2021/10/25 更新

写真a

マエダ タケヤス
前田 健康
MAEDA Takeyasu
所属
教育研究院 医歯学系 歯学系列 教授
歯学部 歯学科 教授
医歯学総合研究科 高度口腔機能教育研究センター 教授
職名
教授
外部リンク

学位

  • 歯学博士 ( 1988年3月   新潟大学 )

研究キーワード

  • 歯学教育

  • 歯科再生学

  • 口腔発生学

  • 口腔組織学

  • 口腔解剖学

研究分野

  • ライフサイエンス / 口腔再生医学、歯科医用工学

  • ライフサイエンス / 解剖学

  • ライフサイエンス / 常態系口腔科学

  • ライフサイエンス / 病態系口腔科学

経歴(researchmap)

  • ワシントン大学   医学部麻酔学講座   客員研究員

    1994年4月 - 1995年2月

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経歴

  • 新潟大学   医歯学総合研究科 口腔生命科学専攻 歯学教育開発室   教授

    2018年8月 - 現在

  • 新潟大学   医歯学総合研究科 高度口腔機能教育研究センター   教授

    2018年8月 - 現在

  • 新潟大学   医歯学総合研究科 高度口腔機能教育研究センター   教授

    2015年6月 - 2018年7月

  • 新潟大学   歯学部 歯学科   教授

    2001年4月 - 現在

  • 新潟大学   医歯学総合研究科   教授

    2001年4月 - 2015年5月

  • 新潟大学   歯学部   教授

    1996年11月 - 2001年3月

  • 新潟大学   歯学部   助教授

    1992年4月 - 1996年1月

  • 新潟大学   歯学部   講師

    1991年11月 - 1992年3月

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学歴

  • 新潟大学   歯学研究科   歯学基礎系

    1984年4月 - 1988年3月

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    国名: 日本国

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  • 新潟大学   歯学部   歯学科

    1978年4月 - 1984年3月

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    国名: 日本国

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所属学協会

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委員歴

  • Japanese dental education association   Chair, committie of international promotion  

    2002年   

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    団体区分:学協会

    Japanese dental education association

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  • 日本歯科教育学会   国際化推進委員会委員長  

    2002年   

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    団体区分:学協会

    日本歯科教育学会

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  • International Association for Denta reserach   Editorial board  

    1998年 - 2001年   

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    団体区分:学協会

    International Association for Denta reserach

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  • 国際歯学研究学会   編集委員  

    1998年 - 2001年   

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    団体区分:学協会

    国際歯学研究学会

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  • 歯科基礎医学会   評議員  

    1996年   

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    団体区分:学協会

    歯科基礎医学会

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  • 顕微鏡学会   評議員  

    1996年   

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    団体区分:学協会

    顕微鏡学会

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  • 新潟歯学会   評議員  

    1996年   

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    団体区分:学協会

    新潟歯学会

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  • 解剖学会   評議員  

    1996年   

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    団体区分:学協会

    解剖学会

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  • Niigata dental society   council  

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    団体区分:学協会

    Niigata dental society

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  • Association for micriscopy   council  

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    団体区分:学協会

    Association for micriscopy

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  • Japanese Association for Oral Biology   Council  

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    団体区分:学協会

    Japanese Association for Oral Biology

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取得資格

  • 歯科医師

 

論文

  • A Repeated State of Acidification Enhances the Anticariogenic Biofilm Activity of Glass Ionomer Cement Containing Fluoro-Zinc-Silicate Fillers. 国際誌

    Traithawit Naksagoon, Shoji Takenaka, Ryoko Nagata, Maki Sotozono, Tatsuya Ohsumi, Takako Ida, Naoki Edanami, Takeyasu Maeda, Yuichiro Noiri

    Antibiotics (Basel, Switzerland)   10 ( 8 )   2021年8月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    This study aimed to evaluate the anticariogenic biofilm activity of a novel zinc-containing glass ionomer cement, Caredyne Restore (CR), using a flow-cell system that reproduces Stephan responses. Streptococcus mutans biofilms were cultured on either CR or hydroxyapatite (HA) discs mounted on a modified Robbins device. The media were allowed to flow at a speed of 2 mL/min for 24 h while exposed to an acidic buffer twice for 30 min to mimic dietary uptake. Acid exposure enhanced biofilm inhibition in the CR group, which showed 2.6 log CFU/mm2 in viable cells and a 2 log copies/mL reduction in total cells compared to the untreated group after 24 h of incubation, suggesting enhanced anticariogenic activity due to the release of fluoride and zinc ions. However, there was no difference in the number of viable and total cells between the two experimental groups after 24 h of incubation in the absence of an acidic environment. The anticariogenic biofilm activity of CR occurs in acidic oral environments, for example in the transient pH drop following dietary uptake. CR restorations are recommended in patients at high risk of caries due to hyposalivation, difficulty brushing, and frequent sugar intake.

    DOI: 10.3390/antibiotics10080977

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  • Perivascular Hedgehog responsive cells play a critical role in peripheral nerve regeneration via controlling angiogenesis. 国際誌

    Yurie Yamada, Jun Nihara, Supaluk Trakanant, Takehisa Kudo, Kenji Seo, Izumi Iida, Kenji Izumi, Masayuki Kurose, Yutaka Shimomura, Miho Terunuma, Takeyasu Maeda, Atsushi Ohazama

    Neuroscience research   2021年6月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Hh signaling has been shown to be activated in intact and injured peripheral nerve. However, the role of Hh signaling in peripheral nerve is not fully understood. In the present study, we observed that Hh signaling responsive cells [Gli1(+) cells] in both the perineurium and endoneurium. In the endoneurium, Gli1(+) cells were classified as blood vessel associated or non-associated. After injury, Gli1(+) cells around blood vessels mainly proliferated to then accumulate into the injury site along with endothelial cells. Hh signaling activity was retained in Gli1(+) cells during nerve regeneration. To understand the role of Hedgehog signaling in Gli1(+) cells during nerve regeneration, we examined mice with Gli1(+) cells-specific inactivation of Hh signaling (Smo cKO). After injury, Smo cKO mice showed significantly reduced numbers of accumulated Gli1(+) cells along with disorganized vascularization at an early stage of nerve regeneration, which subsequently led to an abnormal extension of the axon. Thus, Hh signaling in Gli1(+) cells appears to be involved in nerve regeneration through controlling new blood vessel formation at an early stage.

    DOI: 10.1016/j.neures.2021.06.003

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  • Expression of R-spondins/Lgrs in development of movable craniofacial organs. 国際誌

    Jun Nihara, Maiko Kawasaki, Katsushige Kawasaki, Akane Yamada, Fumiya Meguro, Takehisa Kudo, Supaluk Trakanant, Takahiro Nagai, Isao Saito, Takeyasu Maeda, Atsushi Ohazama

    Gene expression patterns : GEP   41   119195 - 119195   2021年6月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Wnt signaling plays a critical role in the development of many organs, including the major movable craniofacial organs tongue, lip, and eyelid. Four members of the R-spondin family (Rspo1-4) bind to Lgr4/5/6 to regulate the activation of Wnt signaling. However, it is not fully understood how Rspos/Lgrs regulate Wnt signaling during the development of movable craniofacial organs. To address this question, we examined the expression of Rspos, Lgrs, and Axin2 (major mediator of canonical Wnt signaling) during tongue, lip, and eyelid development. The expression of Axin2, Rspos and Lgrs was observed in many similar regions, suggesting that Rspos likely activate canonical Wnt signaling through the Lgr-dependent pathway in these regions. Lgr expression was not detected in regions where Axin2 and Rspos were expressed, suggesting that Rspos might activate canonical Wnt signaling through the Lgr-independent pathway in these regions. In addition, the expression of Rspos and Lgrs were observed in some other regions where Axin2 was not expressed, suggesting the possibility that Rspos and/or Lgrs are involved in non-canonical Wnt signaling or the Wnt-independent pathway. Thus, we identified a dynamic spatiotemporal expression pattern of Rspos and Lgrs during the development of the eyelid, tongue, and lip.

    DOI: 10.1016/j.gep.2021.119195

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  • Rapid elevation of brain-derived neurotrophic factor production in the bilateral trigeminal ganglia by unilateral transection of the mental nerve in mice. 国際誌

    Maiko Yamazaki, Tatsuru Tsurumaki, Yurie Yamada, Takeyasu Maeda, Kenji Seo

    Neuroreport   32 ( 8 )   659 - 665   2021年5月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    OBJECTIVES: Previous spinal nerve injury studies have reported brain-derived neurotrophic factor (BDNF) mRNA upregulation in either the ipsilateral dorsal root ganglion (DRG) neurons or both the contralateral and ipsilateral DRG neurons from early period after peripheral nerve injury. This BDNF elevation induces hyperalgesia in the injured and/or uninjured sites, but this detailed mechanism remains unknown. This study aimed to investigate the BDNF mRNA expression in bilateral DRG neurons caused by unilateral nerve injury and to explore the possible mechanisms by which nitric oxide (NO) mediates BDNF production in the DRG, resulting in contralateral hyperalgesia. METHODS: Early changes in BDNF mRNA expression in the bilateral trigeminal ganglia, within 1 day after mental nerve transection, were examined. Additionally, the effects on BDNF production of the NO synthase inhibitor N(ω)-nitro-l-arginine methyl ester (L-NAME) were investigated in the bilateral trigeminal ganglia. The relationship between injured neurons and BDNF production in the trigeminal ganglia was then assessed using immunohistochemical and retrograde tracing methods. RESULTS: Reverse transcription-PCR analysis demonstrated that unilateral transection of the mental nerve induced a rapid elevation of BDNF mRNA expression, which was inhibited by the intracerebroventricular administration of L-NAME prior to nerve transection. This effect was observed in both the ipsilateral and contralateral sides to the nerve transection. BDNF immunostaining combined with FluoroGold retrograde tracing revealed two types of BDNF-reactive neurons, FluoroGold-labelled and non-FluoroGold-labelled neurons, in the ipsilateral and contralateral sides of the trigeminal ganglia. BDNF-positive cells were also observed in the trigeminal ganglia of other trigeminal nerve branches. CONCLUSIONS: Unilateral nerve injury upregulates BDNF production in the bilateral trigeminal ganglia by NO-mediated and/or indirect activation of afferent neurons, resulting in contralateral hyperalgesia.

    DOI: 10.1097/WNR.0000000000001635

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  • Effects of Erythromycin on Osteoclasts and Bone Resorption via DEL-1 Induction in Mice

    Hikaru Tamura, Tomoki Maekawa, Hisanori Domon, Takumi Hiyoshi, Satoru Hirayama, Toshihito Isono, Karin Sasagawa, Daisuke Yonezawa, Naoki Takahashi, Masataka Oda, Takeyasu Maeda, Koichi Tabeta, Yutaka Terao

    Antibiotics   10 ( 3 )   312 - 312   2021年3月

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    掲載種別:研究論文(学術雑誌)   出版者・発行元:{MDPI} {AG}  

    DOI: 10.3390/antibiotics10030312

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  • MicroRNAs regulate distal region of mandibular development through Hh signaling. 査読 国際誌

    Supaluk Trakanant, Jun Nihara, Takahiro Nagai, Maiko Kawasaki, Katsushige Kawasaki, Yoko Ishida, Fumiya Meguro, Takehisa Kudo, Akane Yamada, Takeyasu Maeda, Isao Saito, Atsushi Ohazama

    Journal of anatomy   238 ( 3 )   711 - 719   2021年3月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Mandibular anomalies are often seen in various congenital diseases, indicating that mandibular development is under strict molecular control. Therefore, it is crucial to understand the molecular mechanisms involved in mandibular development. MicroRNAs (miRNAs) are noncoding small single-stranded RNAs that play a critical role in regulating the level of gene expression. We found that the mesenchymal conditional deletion of miRNAs arising from a lack of Dicer (an essential molecule for miRNA processing, Dicerfl/fl ;Wnt1Cre), led to an abnormal groove formation at the distal end of developing mandibles. At E10.5, when the region forms, inhibitors of Hh signaling, Ptch1 and Hhip1 showed increased expression at the region in Dicer mutant mandibles, while Gli1 (a major mediator of Hh signaling) was significantly downregulated in mutant mandibles. These suggest that Hh signaling was downregulated at the distal end of Dicer mutant mandibles by increased inhibitors. To understand whether the abnormal groove formation inDicer mutant mandibles was caused by the downregulation of Hh signaling, mice with a mesenchymal deletion of Hh signaling activity arising from a lack of Smo (an essential molecule for Hh signaling activation, Smofl/fl ;Wnt1Cre) were examined. Smofl/fl ;Wnt1Cre mice showed a similar phenotype in the distal region of their mandibles to those in Dicerfl/fl ;Wnt1Cre mice. We also found that approximately 400 miRNAs were expressed in wild-type mandibular mesenchymes at E10.5, and six microRNAs were identified as miRNAs with binding potential against both Ptch1 and Hhip1. Their expressions at the distal end of the mandible were confirmed by in situ hybridization. This indicates that microRNAs regulate the distal part of mandibular formation at an early stage of development by involving Hh signaling activity through controlling its inhibitor expression level.

    DOI: 10.1111/joa.13328

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  • Reptinは胎仔上皮におけるDNA損傷応答を介して器官形成を制御する

    目黒 史也, 柿原 嘉人, 川崎 真依子, 川崎 勝盛, 丹原 惇, トゥラカナン・スッパラック, 工藤 武久, 山田 茜, 前田 健康, 多部田 康一, 佐伯 万騎男, 大峡 淳

    新潟歯学会雑誌   50 ( 2 )   115 - 116   2020年12月

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    記述言語:日本語   出版者・発行元:新潟歯学会  

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  • Effect of water aging on the anti-biofilm properties of glass ionomer cement containing fluoro-zinc-silicate fillers. 国際誌

    Traithawit Naksagoon, Tatsuya Ohsumi, Shoji Takenaka, Ryoko Nagata, Taisuke Hasegawa, Takeyasu Maeda, Yuichiro Noiri

    Biofouling   36 ( 9 )   1090 - 1099   2020年10月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    A previous study has reported that a novel fluoro-zinc-silicate glass ionomer cement (Caredyne Restore) showed superior anti-biofilm effects by interfering with bacterial adhesion. However, the active ions may degrade with time. This study aimed to assess the valid anti-biofilm effects of Caredyne Restore after being aged by water immersion for 3 weeks. Streptococcus mutans biofilm was allowed to grow on the surface before and after water aging for 24 h using a modified Robbins device flow-cell system. The results showed water aging promoted biofilm formation. Insufficient amount of fluoride and zinc ions were released from Caredyne Restore after water aging under neutral pH condition. An acidic pH is needed to exert effective anti-biofilm properties. As the release of active ions from Caredyne Restore will gradually decrease after the restoration,  the restoration may not prevent biofilm formation after 3 weeks while neutral pH is maintained by the buffering capacity of saliva.

    DOI: 10.1080/08927014.2020.1856371

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  • Overactivation of the NF-κB pathway impairs molar enamel formation. 査読 国際誌

    Akane Yamada, Maiko Kawasaki, Yasuo Miake, Yurie Yamada, James Blackburn, Katsushige Kawasaki, Supaluk Trakanant, Takahiro Nagai, Jun Nihara, Takehisa Kudo, Fumiya Meguro, Ruth Schmidt-Ullrich, Bigang Liu, Yinling Hu, Angustias Page, Ángel Ramírez, Paul T Sharpe, Takeyasu Maeda, Ritsuo Takagi, Atsushi Ohazama

    Oral diseases   26 ( 7 )   1513 - 1522   2020年10月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    OBJECTIVE: Hypohidrotic ectodermal dysplasia (HED) is a hereditary disorder characterized by abnormal structures and functions of the ectoderm-derived organs, including teeth. HED patients exhibit a variety of dental symptoms, such as hypodontia. Although disruption of the EDA/EDAR/EDARADD/NF-κB pathway is known to be responsible for HED, it remains unclear whether this pathway is involved in the process of enamel formation. EXPERIMENTAL SUBJECTS AND METHODS: To address this question, we examined the mice overexpressing Ikkβ (an essential component required for the activation of NF-κB pathway) under the keratin 5 promoter (K5-Ikkβ). RESULTS: Upregulation of the NF-κB pathway was confirmed in the ameloblasts of K5-Ikkβ mice. Premature abrasion was observed in the molars of K5-Ikkβ mice, which was accompanied by less mineralized enamel. However, no significant changes were observed in the enamel thickness and the pattern of enamel rods in K5-Ikkβ mice. Klk4 expression was significantly upregulated in the ameloblasts of K5-Ikkβ mice at the maturation stage, and the expression of its substrate, amelogenin, was remarkably reduced. This suggests that abnormal enamel observed in K5-Ikkβ mice was likely due to the compromised degradation of enamel protein at the maturation stage. CONCLUSION: Therefore, we could conclude that the overactivation of the NF-κB pathway impairs the process of amelogenesis.

    DOI: 10.1111/odi.13384

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  • Drug-Induced Naïve iPS Cells Exhibit Better Performance than Primed iPS Cells with Respect to the Ability to Differentiate into Pancreatic β-Cell Lineage. 査読 国際誌

    Yuki Kiyokawa, Masahiro Sato, Hirofumi Noguchi, Emi Inada, Yoko Iwase, Naoko Kubota, Tadashi Sawami, Miho Terunuma, Takeyasu Maeda, Haruaki Hayasaki, Issei Saitoh

    Journal of clinical medicine   9 ( 9 )   2020年9月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Pluripotent stem cells are classified as naïve and primed cells, based on their in vitro growth characteristics and potential to differentiate into various types of cells. Human-induced pluripotent stem cells (iPSCs, also known as epiblast stem cells [EpiSCs]) have limited capacity to differentiate and are slightly more differentiated than naïve stem cells (NSCs). Although there are several in vitro protocols that allow iPSCs to differentiate into pancreatic lineage, data concerning generation of β-cells from these iPSCs are limited. Based on the pluripotentiality of NSCs, it was hypothesized that NSCs can differentiate into pancreatic β-cells when placed under an appropriate differentiation induction condition. We examined whether NSCs can be efficiently induced to form potentially pancreatic β cells after being subjected to an in vitro protocol. Several colonies resembling in vitro-produced β-cell foci, with β-cell-specific marker expression, were observed when NSC-derived embryoid bodies (EBs) were induced to differentiate into β-cell lineage. Conversely, EpiSC-derived EBs failed to form such foci in vitro. Intrapancreatic grafting of the in vitro-formed β-cell foci into nude mice (BALB/c-nu/nu) generated a cell mass containing insulin-producing cells (IPCs), without noticeable tumorigenesis. These NSCs can be used as a promising resource for curing type 1 diabetes.

    DOI: 10.3390/jcm9092838

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  • Erythromycin inhibits neutrophilic inflammation and mucosal disease by upregulating DEL-1. 査読 国際誌

    Tomoki Maekawa, Hikaru Tamura, Hisanori Domon, Takumi Hiyoshi, Toshihito Isono, Daisuke Yonezawa, Naoki Hayashi, Naoki Takahashi, Koichi Tabeta, Takeyasu Maeda, Masataka Oda, Athanasios Ziogas, Vasileia Ismini Alexaki, Triantafyllos Chavakis, Yutaka Terao, George Hajishengallis

    JCI insight   5 ( 15 )   2020年8月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:American Society for Clinical Investigation  

    Macrolide antibiotics exert antiinflammatory effects; however, little is known regarding their immunomodulatory mechanisms. In this study, using 2 distinct mouse models of mucosal inflammatory disease (LPS-induced acute lung injury and ligature-induced periodontitis), we demonstrated that the antiinflammatory action of erythromycin (ERM) is mediated through upregulation of the secreted homeostatic protein developmental endothelial locus-1 (DEL-1). Consistent with the anti-neutrophil recruitment action of endothelial cell-derived DEL-1, ERM inhibited neutrophil infiltration in the lungs and the periodontium in a DEL-1-dependent manner. Whereas ERM (but not other antibiotics, such as josamycin and penicillin) protected against lethal pulmonary inflammation and inflammatory periodontal bone loss, these protective effects of ERM were abolished in Del1-deficient mice. By interacting with the growth hormone secretagogue receptor and activating JAK2 in human lung microvascular endothelial cells, ERM induced DEL-1 transcription that was mediated by MAPK p38 and was CCAAT/enhancer binding protein-β dependent. Moreover, ERM reversed IL-17-induced inhibition of DEL-1 transcription, in a manner that was dependent not only on JAK2 but also on PI3K/AKT signaling. Because DEL-1 levels are severely reduced in inflammatory conditions and with aging, the ability of ERM to upregulate DEL-1 may lead to a novel approach for the treatment of inflammatory and aging-related diseases.

    DOI: 10.1172/jci.insight.136706

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  • Gli3 is a Key Factor in the Schwann Cells from Both Intact and Injured Peripheral Nerves. 査読 国際誌

    Yurie Yamada, Supaluk Trakanant, Jun Nihara, Takehisa Kudo, Kenji Seo, Makio Saeki, Masayuki Kurose, Daisuke Matsumaru, Takeyasu Maeda, Atsushi Ohazama

    Neuroscience   432   229 - 239   2020年4月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Hedgehog (Hh) signaling has been shown to be involved in regulating both intact and injured peripheral nerves. Therefore, it is critical to understand how Hh signaling is regulated in the peripheral nerve. One of the transcription factors of the Hh signaling pathway, Gli3, functions as both a repressor and an activator of Hh signaling activity. However, it remains unclear whether Gli3 is involved in controlling the intact and/or injured peripheral nerves. We found that Gli3 act as a repressor in the Schwann cells (SCs) of intact sciatic nerves. Although Dhh and Ptch1 expression were present, Hh signaling was not activated in these SCs. Moreover, heterozygous Gli3 mutation (Gli3-/+) induced ectopic Hh signaling activity in SCs. Hh signaling was thus suppressed by Gli3 in the SCs of intact sciatic nerves. Minor morphological changes were observed in the intact nerves from Gli3-/+ mice. Gli3 expression was significantly decreased following injury and ligand expression switched from Dhh to Shh, which activated Hh signaling in SCs from wild-type mice. Changes of these ligands was found to be important for nerve regeneration in which the downregulation of Gli3 was also involved. In fact, Gli3-/+ mice exhibited accelerated ligand switching and subsequent nerve regeneration. Both suppression of Hh signaling with Gli3 in the intact nerves and activation of Hh signaling without Gli3 in the injured nerve were observed in the SCs in an autocrine manner. Thus, Gli3 is a key factor in the control of intact peripheral nerve homeostasis and nerve regeneration.

    DOI: 10.1016/j.neuroscience.2020.02.036

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  • Molecular mechanisms in palatal rugae development. 査読 国際誌

    Supaluk Trakanant, Jun Nihara, Maiko Kawasaki, Fumiya Meguro, Akane Yamada, Katsushige Kawasaki, Isao Saito, Maeda Takeyasu, Atsushi Ohazama

    Journal of oral biosciences   62 ( 1 )   30 - 35   2020年3月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    BACKGROUND: Periodic patterning of iterative structures is diverse across the animal kingdom. Clarifying the molecular mechanisms involved in the formation of these structures helps to elucidate the genetic commonality of developmental processes, as organs with these structures are believed to share the same molecular mechanisms and fundamental processes. Palatal rugae are periodic corrugated structures on the hard palate and are conserved in all mammals. Although the numbers and patterns of the palatal rugae are species specific, they are consistent in each mammalian species, except humans. HIGHLIGHT: Palatal rugae development is thus under strict genetic control in most mammals and is an excellent model to investigate the genetic commonality of developmental processes to form periodic patterning. CONCLUSION: This review highlights the current understanding of the molecular mechanisms of palatal rugae development.

    DOI: 10.1016/j.job.2019.12.002

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  • Effect of a novel glass ionomer cement containing fluoro-zinc-silicate fillers on biofilm formation and dentin ion incorporation. 査読 国際誌

    Taisuke Hasegawa, Shoji Takenaka, Tatsuya Ohsumi, Takako Ida, Hayato Ohshima, Yutaka Terao, Traithawit Naksagoon, Takeyasu Maeda, Yuichiro Noiri

    Clinical oral investigations   24 ( 2 )   963 - 970   2020年2月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:SPRINGER HEIDELBERG  

    OBJECTIVES: This study is aimed at evaluating the effect of a new glass ionomer cement (GIC) containing fluoro-zinc-silicate fillers on biofilm formation and ion incorporation. MATERIALS AND METHODS: Streptococcus mutans biofilms were developed on two GIC materials: Caredyne Restore (CD) and Fuji VII (FJ); and hydroxyapatite (HA) for 24 h at 37 °C using a flow cell system. The morphological structure and bacterial viability were analyzed using a confocal laser scanning microscopy. Bacterial adhesion during the initial 2 h was also assessed by viable cell counting. To study the ion incorporation, restored cavities prepared on the root surfaces of human incisors were subjected to the elemental mapping of the zinc and fluoride ions in the GIC-dentin interface using a wavelength-dispersive X-ray spectroscopy electron probe microanalyzer. RESULTS: Morphological observations revealed that biofilm formation in the CD group was remarkably inhibited compared with the HA and FJ groups, exhibiting sparse, thinner biofilm clusters. The microorganisms adhering to the CD group were significantly inhibited, revealing 2.9 ± 0.4 for CD, 4.9 ± 0.2 for FJ, and 5.4 ± 0.4 log colony-forming units (CFU) for HA. The CD zinc ion incorporation depth was 72.2 ± 8.0 μm. The fluoride penetration of CD was three times deeper than that of FJ; this difference was statistically significant (p < 0.05). CONCLUSIONS: Enhanced by the incorporation of zinc and fluoride ions, the new GIC inhibited biofilm formation by interfering with bacterial adhesion. CLINICAL RELEVANCE: A novel GIC comprised of fluoro-zinc-silicate fillers may improve clinical outcomes, such as root caries and minimally invasive dentistry.

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  • Ift88 is involved in mandibular development. 査読 国際誌

    Atsushi Kitamura, Maiko Kawasaki, Katsushige Kawasaki, Fumiya Meguro, Akane Yamada, Takahiro Nagai, Yasumitsu Kodama, Supaluk Trakanant, Paul T Sharpe, Takeyasu Maeda, Ritsuo Takagi, Atsushi Ohazama

    Journal of anatomy   236 ( 2 )   317 - 324   2020年2月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    The mandible is a crucial organ in both clinical and biological fields due to the high frequency of congenital anomalies and the significant morphological changes during evolution. Primary cilia play a critical role in many biological processes, including the determination of left/right axis patterning, the regulation of signaling pathways, and the formation of bone and cartilage. Perturbations in the function of primary cilia are known to cause a wide spectrum of human diseases: the ciliopathies. Craniofacial dysmorphologies, including mandibular deformity, are often seen in patients with ciliopathies. Mandibular development is characterized by chondrogenesis and osteogenesis; however, the role of primary cilia in mandibular development is not fully understood. To address this question, we generated mice with mesenchymal deletions of the ciliary protein, Ift88 (Ift88fl/fl ;Wnt1Cre). Ift88fl/fl ;Wnt1Cre mice showed ectopic mandibular bone formation, whereas Ift88 mutant mandible was slightly shortened. Meckel's cartilage was modestly expanded in Ift88fl/fl ;Wnt1Cre mice. The downregulation of Hh signaling was found in most of the mesenchyme of Ift88 mutant mandible. However, mice with a mesenchymal deletion of an essential molecule for Hh signaling activity, Smo (Smofl/fl ;Wnt1Cre), showed only ectopic mandibular formation, whereas Smo mutant mandible was significantly shortened. Ift88 is thus involved in chondrogenesis and osteogenesis during mandibular development, partially through regulating Sonic hedgehog (Shh) signaling.

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  • M2 Phenotype Macrophages Colocalize with Schwann Cells in Human Dental Pulp 査読

    N. Yoshiba, N. Edanami, N. Ohkura, T. Maekawa, A. Tohma, K. Izumi, T. Maeda, A. Hosoya, H. Nakamura, Y. Noiri, K. Yoshiba

    Journal of Dental Research   99 ( 3 )   329 - 338   2020年1月

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  • Primary cilia in murine palatal rugae development. 査読 国際誌

    Mayuko Nakaniwa, Maiko Kawasaki, Katsushige Kawasaki, Akane Yamada, Fumiya Meguro, Maeda Takeyasu, Atsushi Ohazama

    Gene expression patterns : GEP   34   119062 - 119062   2019年12月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Periodic patterning of iterative structures is a fundamental process during embryonic development, since these structures are diverse across the animal kingdom. Therefore, elucidating the molecular mechanisms in the formation of these structures promotes understanding of the process of organogenesis. Periodically patterned ridges, palatal rugae (situated on the hard palate of mammals), are an excellent experimental model to clarify the molecular mechanisms involved in the formation of periodic patterning of iterative structures. Primary cilia are involved in many biological events, including the regulation of signaling pathways such as Shh and non-canonical Wnt signaling. However, the role of primary cilia in the development of palatal rugae remains unclear. We found that primary cilia were localized to the oral cavity side of the interplacode epithelium of the palatal rugae, whereas restricted localization of primary cilia could not be detected in other regions. Next, we generated mice with a placodal conditional deletion of the primary cilia protein Ift88, using ShhCre mice (Ift88 fl/fl;ShhCre). Highly disorganized palatal rugae were observed in Ift88 fl/fl;ShhCre mice. Furthermore, by comparative in situ hybridization analysis, many Shh and non-canonical Wnt signaling-related molecules showed spatiotemporal expression patterns during palatal rugae development, including restricted expression in the epithelium (placodes and interplacodes) and mesenchyme. Some of these expression were found to be altered in Ift88 fl/fl;ShhCre mice. Primary cilia is thus involved in development of palatal rugae.

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  • Histochemical examination on principal collagen fibers in periodontal ligaments of ascorbic acid-deficient ODS-od/od rats. 査読 国際誌

    Tomoka Hasegawa, Yukina Miyamoto-Takasaki, Miki Abe, Zixuan Qiu, Tomomaya Yamamoto, Taiji Yoshida, Hirona Yoshino, Hiromi Hongo, Ayako Yokoyama, Muneteru Sasaki, Shinichiro Kuroshima, Kuniko Hara, Masatoshi Kobayashi, Yasuhiro Akiyama, Takeyasu Maeda, Paulo Henrique Luiz de Freitas, Minqi Li, Norio Amizuka

    Microscopy (Oxford, England)   68 ( 5 )   349 - 358   2019年10月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    In this study, we aimed to clarify the role of ascorbic acid in collagen synthesis in periodontal ligaments using osteogenic disorder Shionogi (ODS)/ShiJcl-od/od rats lacking L-gulonolactone oxidase. These rats cannot synthesize ascorbic acid in vivo. Eight-week-old ODS/ShiJcl-od/od male rats were administered ascorbic acid solution at a concentration of 200 mg/dL (control group, n = 6) or ascorbic acid solution at concentration of 0.3 mg/dL (insufficient group, n = 12). Six rats of the insufficient group were then given with ascorbic acid solution at concentration of 200 mg/dL for additional 3 weeks (rescued group, n = 6), and then, their mandibles were histochemically examined. Consequently, the insufficient group specimens were seen to possess fewer collagen fibers, and silver impregnation revealed numerous fine, reticular fiber-like fibrils branching off from collagen in the periodontal ligaments. In control group, faint immunoreactivities for matrix metalloproteinase (MMP)2 and cathepsin H were seen in the periphery of blood vessels and throughout the ligament, respectively. In contrast, in the insufficient group, intense MMP2-immunoreactivity was observed to be associated with collagen fibrils in the periodontal ligaments, and cathepsin H-immunopositivity was seen in ligamentous cells. The rescued group showed abundant collagen fibers filling the periodontal ligament space. Under transmission electron microscopy, ligamentous fibroblasts incorporated collagen fibrils into tubular endosomes/lysosomes while simultaneously synthesizing collagen fibril bundles. Thus, ascorbic acid insufficiency affected the immunolocalization of cathepsin H and MMP2; however, ligamentous fibroblasts appear to possess the potential to synthesize collagen fibers when supplied with ascorbic acid.

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  • Bmp signaling in molar cusp formation. 査読 国際誌

    Fumiya Meguro, Thantrira Porntaveetus, Maiko Kawasaki, Katsushige Kawasaki, Akane Yamada, Yoshito Kakihara, Makio Saeki, Koichi Tabeta, John A Kessler, Takeyasu Maeda, Atsushi Ohazama

    Gene expression patterns : GEP   32   67 - 71   2019年6月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Tooth cusp is a crucial structure, since the shape of the molar tooth is determined by number, shape, and size of the cusp. Bone morphogenetic protein (Bmp) signaling is known to play a critical role in tooth development, including in initiation. However, it remains unclear whether Bmp signaling is also involved in cusp formation. To address this question, we examined cusp in two different transgenic mouse lines: mice with overexpression of Bmp4 (K14-Bmp4), and those with Bmp inhibitor, Noggin, (K14-Noggin) under keratin14 (K14) promoter. K14-Noggin mice demonstrated extra cusps, whereas reduced number of cusps was observed in K14-Bmp4 mice. To further understand how Bmps are expressed during cusp formation, we performed whole-mount in situ hybridisation analysis of three major Bmps (Bmp2, Bmp4, and Bmp7) in murine maxillary and mandibular molars from E14.5 to P3. The linear expressions of Bmp2 and Bmp4 were observed in both maxillary and mandibular molars at E14.5. The expression patterns of Bmp2 and Bmp4 became significantly different between the maxillary and mandibular molars at E16.5. At P3, all Bmps were expressed in all the cusp regions of the maxillary molar; however, the patterns differed. All Bmps thus exhibited dynamic temporo-spatial expression during the cusp formation. It could therefore be inferred that Bmp signaling is involved in regulating cusp formation.

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  • Ift88 limits bone formation in maxillary process through suppressing apoptosis. 査読 国際誌

    Momoko Watanabe, Maiko Kawasaki, Katsushige Kawasaki, Atsushi Kitamura, Takahiro Nagai, Yasumitsu Kodama, Fumiya Meguro, Akane Yamada, Paul T Sharpe, Takeyasu Maeda, Ritsuo Takagi, Atsushi Ohazama

    Archives of oral biology   101   43 - 50   2019年5月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    OBJECTIVE: The development of the maxillary bone is under strict molecular control because of its complicated structure. Primary cilia play a critical role in craniofacial development, since defects in primary cilia are known to cause congenital craniofacial dysmorphologies as a wide spectrum of human diseases: the ciliopathies. The primary cilia also are known to regulate bone formation. However, the role of the primary cilia in maxillary bone development is not fully understood. DESIGN: To address this question, we generated mice with a mesenchymal conditional deletion ofIft88 using the Wnt1Cre mice (Ift88fl/fl;Wnt1Cre). The gene Ift88 encodes a protein that is required for the function and formation of primary cilia. RESULTS: It has been shown thatIft88fl/fl;Wnt1Cre mice exhibit cleft palate. Here, we additionally observed excess bone formation in the Ift88 mutant maxillary process. We also found ectopic apoptosis in the Ift88 mutant maxillary process at an early stage of development. To investigate whether the ectopic apoptosis is related to the Ift88 mouse maxillary phenotypes, we generated Ift88fl/fl;Wnt1Cre;p53-/- mutants to reduce apoptosis. The Ift88fl/fl;Wnt1Cre;p53-/- mice showed no excess bone formation, suggesting that the cells evading apoptosis by the presence of Ift88 in wild-type mice limit bone formation in maxillary development. On the other hand, the palatal cleft was retained in the Ift88fl/fl;Wnt1Cre;p53-/- mice, indicating that the excess bone formation or abnormal apoptosis was independent of the cleft palate phenotype in Ift88 mutant mice. CONCLUSIONS: Ift88 limits bone formation in the maxillary process by suppressing apoptosis.

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  • Semaphorin 3A Inhibits Nerve Regeneration During Early Stage after Inferior Alveolar Nerve Transection. 査読 国際誌

    Hiroko Kanemaru, Yurie Yamada, Atsushi Ohazama, Takeyasu Maeda, Kenji Seo

    Scientific reports   9 ( 1 )   4245 - 4245   2019年3月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Neuroma formation at sites of injury can impair peripheral nerve regeneration. Although the involvement of semaphorin 3A has been suggested in neuroma formation, this detailed process after injury is not fully understood. This study was therefore undertaken to examine the effects of semaphorin 3A on peripheral nerve regeneration during the early stage after injury. Immunohistochemistry for semaphorin 3A and PGP9.5, a general neuronal marker, was carried out for clarify chronological changes in their expressions after transection of the mouse inferior alveolar nerve thorough postoperative days 1 to 7. At postoperative day 1, the proximal stump of the damaged IAN exhibited semaphorin 3A, while the distal stump lacked any immunoreactivity. From this day on, its expression lessened, ultimately disappearing completely in all regions of the transected inferior alveolar nerve. A local administration of an antibody to semaphorin 3A into the nerve transection site at postoperative day 3 inhibited axon sprouting at the injury site. This antibody injection increased the number of trigeminal ganglion neurons labeled with DiI (paired t-test, p < 0.05). Immunoreactivity of the semaphorin 3A receptor, neuropilin-1, was also detected at the proximal stump at postoperative day 1. These results suggest that nerve injury initiates semaphorin 3A production in ganglion neurons, which is then delivered through the nerve fibers to the proximal end, thereby contributes to the inhibition of axonal sprouting from the proximal region of injured nerves in the distal direction. To our knowledge, this is the first report to reveal the involvement of Sema3A in the nerve regeneration process at its early stage.

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  • MicroRNAs control eyelid development through regulating Wnt signaling. 査読 国際誌

    Takahiro Nagai, Supaluk Trakanant, Maiko Kawasaki, Katsushige Kawasaki, Yurie Yamada, Momoko Watanabe, James Blackburn, Yoko Otsuka-Tanaka, Mitsue Hishinuma, Atsushi Kitatmura, Fumiya Meguro, Akane Yamada, Yasumitsu Kodama, Takeyasu Maeda, Qiliang Zhou, Yasuo Saijo, Akihiro Yasue, Paul T Sharpe, Robert Hindges, Ritsuo Takagi, Atsushi Ohazama

    Developmental dynamics : an official publication of the American Association of Anatomists   248 ( 3 )   201 - 210   2019年3月

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    記述言語:英語  

    BACKGROUND: The timing, location, and level of gene expression are crucial for normal organ development, because morphogenesis requires strict genetic control. MicroRNAs (miRNAs) are noncoding small single-stranded RNAs that play a critical role in regulating gene expression level. Although miRNAs are known to be involved in many biological events, the role of miRNAs in organogenesis is not fully understood. Mammalian eyelids fuse and separate during development and growth. In mice, failure of this process results in the eye-open at birth (EOB) phenotype. RESULTS: It has been shown that conditional deletion of mesenchymal Dicer (an essential protein for miRNA processing; Dicer fl/fl ;Wnt1Cre) leads to the EOB phenotype with full penetrance. Here, we identified that the up-regulation of Wnt signaling resulted in the EOB phenotype in Dicer mutants. Down-regulation of Fgf signaling observed in Dicer mutants was caused by an inverse relationship between Fgf and Wnt signaling. Shh and Bmp signaling were down-regulated as the secondary effects in Dicer fl/fl ;Wnt1Cre mice. Wnt, Shh, and Fgf signaling were also found to mediate the epithelial-mesenchymal interactions in eyelid development. CONCLUSIONS: miRNAs control eyelid development through Wnt. Developmental Dynamics 248:201-210, 2019. © 2019 Wiley Periodicals, Inc.

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  • A bacterial metabolite ameliorates periodontal pathogen-induced gingival epithelial barrier disruption via GPR40 signaling 査読

    Miki Yamada, Naoki Takahashi, Yumi Matsuda, Keisuke Sato, Mai Yokoji, Benso Sulijaya, Tomoki Maekawa, Tatsuo Ushiki, Yoshikazu Mikami, Manabu Hayatsu, Yusuke Mizutani, Shigenobu Kishino, Jun Ogawa, Makoto Arita, Koichi Tabeta, Takeyasu Maeda, Kazuhisa Yamazaki

    Scientific Reports   8 ( 1 )   9008   2018年12月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:Nature Publishing Group  

    Several studies have demonstrated the remarkable properties of microbiota and their metabolites in the pathogenesis of several inflammatory diseases. 10-Hydroxy-cis-12-octadecenoic acid (HYA), a bioactive metabolite generated by probiotic microorganisms during the process of fatty acid metabolism, has been studied for its protective effects against epithelial barrier impairment in the intestines. Herein, we examined the effect of HYA on gingival epithelial barrier function and its possible application for the prevention and treatment of periodontal disease. We found that GPR40, a fatty acid receptor, was expressed on gingival epithelial cells
    activation of GPR40 by HYA significantly inhibited barrier impairment induced by Porphyromonas gingivalis, a representative periodontopathic bacterium. The degradation of E-cadherin and beta-catenin, basic components of the epithelial barrier, was prevented in a GPR40-dependent manner in vitro. Oral inoculation of HYA in a mouse experimental periodontitis model suppressed the bacteria-induced degradation of E-cadherin and subsequent inflammatory cytokine production in the gingival tissue. Collectively, these results suggest that HYA exerts a protective function, through GPR40 signaling, against periodontopathic bacteria-induced gingival epithelial barrier impairment and contributes to the suppression of inflammatory responses in periodontal diseases.

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  • Peptides from rice endosperm protein restrain periodontal bone loss in mouse model of periodontitis. 査読

    Tamura H, Maekawa T, Domon H, Hiyoshi T, Yonezawa D, Nagai K, Ochiai A, Taniguchi M, Tabeta K, Maeda T, Terao Y

    Arch. Oral Biol.   98   132 - 139   2018年11月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

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  • The Sonic Hedgehog signaling pathway regulates inferior alveolar nerve regeneration. 査読 国際誌

    Yurie Yamada, Atsushi Ohazama, Takeyasu Maeda, Kenji Seo

    Neuroscience letters   671   114 - 119   2018年4月

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    記述言語:英語  

    Activation of Shh signaling is known to be observed following injury of the peripheral nerves such as the sciatic nerve. However, the precise role of Shh signaling during peripheral nerve regeneration is not fully understood. The inferior alveolar nerve (IAN) is most commonly injured during oral surgery. Unlike the sciatic nerve, the IAN is isolated from other craniofacial tissues, as it resides in a long bony canal within the mandible. The IAN is thus an excellent experimental model for investigating peripheral nerve regeneration. In this study, the role of Shh signaling in peripheral nerve regeneration was investigated using the mouse IAN transection model. During regeneration, Shh signaling was activated within the entire distal region of the IAN and proximal stumps. Inhibition of Shh signaling by cyclopamine application at the transection site led to abnormal axon growth in random directions, a reduced number of macrophages, and an increase in myelin debris within the distal region. Shh signaling is thus involved in peripheral nerve regeneration via the regulation of myelin degradation.

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  • Sox Genes Show Spatiotemporal Expression during Murine Tongue and Eyelid Development. 査読 国際誌

    Ryuichi Ishikawa, Maiko Kawasaki, Katsushige Kawasaki, Akane Yamada, Supaluk Trakanant, Fumiya Meguro, Atsushi Kitamura, Takehisa Kudo, Takeyasu Maeda, Atsushi Ohazama

    International journal of dentistry   2018   1601363 - 1601363   2018年

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    The tongue is a critical organ, involved in functions such as speaking, swallowing, mastication, and degustation. Although Sox genes are known to play critical roles in many biological processes, including organogenesis, the expression of the Sox family members during tongue development remains unclear. We therefore performed a comparative in situ hybridization analysis of 17 Sox genes (Sox1-14, 17, 18, and 21) during murine tongue development. Sox2, 4, 6, 8, 9, 10, 11, 12, and 21 were found to be expressed in the tongue epithelium, whereas Sox2, 4-6, 8-11, 13, and 21 showed expression in the mesenchyme of the developing tongue. Expression of Sox1, 4, 6, 8-12, and 21 were observed in the developing tongue muscle. Sox5 and 13 showed expression only at E12, while Sox1 expression was observed only on E18. Sox6, 8, 9, and 12 showed expression at several stages. Although the expression of Sox2, 4, 10, 11, and 21 was detected during all the four stages of tongue development, their expression patterns differed among the stages. We thus identified a dynamic spatiotemporal expression pattern of the Sox genes during murine tongue development. To understand whether Sox genes are involved in the development of other craniofacial organs through similar roles to those in tongue development, we also examined the expression of Sox genes in eyelid primordia, which also contain epithelium, mesenchyme, and muscle. However, expression patterns and timing of Sox genes differed between tongue and eyelid development. Sox genes are thus related to organogenesis through different functions in each craniofacial organ.

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  • Lrp4/Wise regulates palatal rugae development through Turing-type reaction-diffusion mechanisms. 査読 国際誌

    Maiko Kawasaki, Katsushige Kawasaki, Fumiya Meguro, Akane Yamada, Ryuichi Ishikawa, Thantrira Porntaveetus, James Blackburn, Yoko Otsuka-Tanaka, Naoaki Saito, Masato S Ota, Paul T Sharpe, John A Kessler, Joachim Herz, Martyn T Cobourne, Takeyasu Maeda, Atsushi Ohazama

    PloS one   13 ( 9 )   e0204126   2018年

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    記述言語:英語  

    Periodic patterning of iterative structures is diverse across the animal kingdom. Clarifying the molecular mechanisms involved in the formation of these structure helps to elucidate the process of organogenesis. Turing-type reaction-diffusion mechanisms have been shown to play a critical role in regulating periodic patterning in organogenesis. Palatal rugae are periodically patterned ridges situated on the hard palate of mammals. We have previously shown that the palatal rugae develop by a Turing-type reaction-diffusion mechanism, which is reliant upon Shh (as an inhibitor) and Fgf (as an activator) signaling for appropriate organization of these structures. The disturbance of Shh and Fgf signaling lead to disorganized palatal rugae. However, the mechanism itself is not fully understood. Here we found that Lrp4 (transmembrane protein) was expressed in a complementary pattern to Wise (a secreted BMP antagonist and Wnt modulator) expression in palatal rugae development, representing Lrp4 expression in developing rugae and Wise in the inter-rugal epithelium. Highly disorganized palatal rugae was observed in both Wise and Lrp4 mutant mice, and these mutants also showed the downregulation of Shh signaling, which was accompanied with upregulation of Fgf signaling. Wise and Lrp4 are thus likely to control palatal rugae development by regulating reaction-diffusion mechanisms through Shh and Fgf signaling. We also found that Bmp and Wnt signaling were partially involved in this mechanism.

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  • Vascularization via activation of VEGF-VEGFR signaling is essential for peripheral nerve regeneration. 査読

    Yohei Nishida, Yurie Yamada, Hiroko Kanemaru, Atsushi Ohazama, Takeyasu Maeda, Kenji Seo

    Biomedical research (Tokyo, Japan)   39 ( 6 )   287 - 294   2018年

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Peripheral nerve injury has been suggested to up-regulate mRNA for the vascular endothelial growth factor (VEGF) which enhances nerve regeneration. VEGF is known to regulate angiogenesis by binding with a specific receptor, the vascular endothelial growth factor receptor (VEGFR). However, little is known about the involvement of VEGF-VEGFR signaling in the nerve regeneration at early stages though previous studies contained a lengthy observation. The present study examined that relationship between angiogenesis and peripheral nerve regeneration at the early stage after nerve transection by focusing on the chronological changes in the expression patterns of VEGF-VEGFR signaling. This study used our previously reported experimental model for nerve regeneration following the transection of the inferior alveolar nerve (IAN) in mice. In a double staining of PGP9.5 and CD31, respective markers for the nerve fibers and endothelial cells, CD31 immunoreactions first appeared in the injury site on postoperative (PO) day 2 when the transected nerve fibers had not been re-connected. The most intense immunoreaction for CD31 was found around the regenerating nerve fibers extending from the proximal stump on PO day 3, but it gradually lessened to disappear by PO day 7. The expression patterns of VEGFR1 and VEGFR2 showed similar chronological changes through the observation periods, with most intense immunoreaction found on PO day 3. Western blotting of total protein extracted from the injury site demonstrated the clear bands for VEGF-A and VEGF-B on PO day 2, indicating a time lag for the expression of ligands and receptors. A local administration of antibody to VEGF-A inhibited the elongation of the nerve fibers from the proximal stump. Furthermore, this administration of VEGF-A antibody inhibited the expression of CD31 in the gap between proximal and distal stumps. These results indicated that a nerve injury initiates productions in VEGF-A and VEFG-B, followed with the expression of VEGFR1 and VEGFR2 at early stages after the nerve injury. Taken these findings together, it is reasonable to postulate that immediate response of VEGF-VEGFR signaling to nerve injury plays a crucial role in local angiogenesis, resulting in a trigger for the regeneration of the nerve fibers in mouse IAN.

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  • Sox Genes Show Spatiotemporal Expression during Murine Tongue and Eyelid Development. 査読 国際誌

    Ryuichi Ishikawa, Maiko Kawasaki, Katsushige Kawasaki, Akane Yamada, Supaluk Trakanant, Fumiya Meguro, Atsushi Kitamura, Takehisa Kudo, Takeyasu Maeda, Atsushi Ohazama

    International journal of dentistry   2018   1601363 - 1601363   2018年

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    記述言語:英語  

    The tongue is a critical organ, involved in functions such as speaking, swallowing, mastication, and degustation. Although Sox genes are known to play critical roles in many biological processes, including organogenesis, the expression of the Sox family members during tongue development remains unclear. We therefore performed a comparative in situ hybridization analysis of 17 Sox genes (Sox1-14, 17, 18, and 21) during murine tongue development. Sox2, 4, 6, 8, 9, 10, 11, 12, and 21 were found to be expressed in the tongue epithelium, whereas Sox2, 4-6, 8-11, 13, and 21 showed expression in the mesenchyme of the developing tongue. Expression of Sox1, 4, 6, 8-12, and 21 were observed in the developing tongue muscle. Sox5 and 13 showed expression only at E12, while Sox1 expression was observed only on E18. Sox6, 8, 9, and 12 showed expression at several stages. Although the expression of Sox2, 4, 10, 11, and 21 was detected during all the four stages of tongue development, their expression patterns differed among the stages. We thus identified a dynamic spatiotemporal expression pattern of the Sox genes during murine tongue development. To understand whether Sox genes are involved in the development of other craniofacial organs through similar roles to those in tongue development, we also examined the expression of Sox genes in eyelid primordia, which also contain epithelium, mesenchyme, and muscle. However, expression patterns and timing of Sox genes differed between tongue and eyelid development. Sox genes are thus related to organogenesis through different functions in each craniofacial organ.

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  • The effect of bone mass and architecture on mandibular condyle after mandibular distraction 査読

    Daisuke Suda, Atsushi Ohazama, Takeyasu Maeda, Tadaharu Kobayashi

    ORAL SURGERY ORAL MEDICINE ORAL PATHOLOGY ORAL RADIOLOGY   124 ( 4 )   339 - 347   2017年10月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:ELSEVIER SCIENCE INC  

    Objective. Mandibular distraction surgery is a critical treatment for jaw deformity. However, abnormal mandibular condylar bone resorption is often seen as complication after surgery. Our previous study using a rat mandibular distraction model suggested that overloading leads to mandibular condylar resorption. Host factors are also believed to influence the resorption. To understand the relationship between host factors and resorption, we investigated the effect of changing bone mass and architecture on the mandibular condyle using FK506.
    Study Design. FK506, an immunosuppressant, was used to compromise bone mass and architecture in this study. Animals were divided into 4 groups: distraction surgery (Dist), FK506 administration (FK), distraction surgery with FK506 administration (FK + Dist), and no surgery or FK506 administration (Cont).
    Results. The FK group showed reduced bone mass and impaired bone architecture. The Dist group exhibited abnormal bone resorption on the surface of the condyles, which was slightly exacerbated in the FK + Dist group. Bone defect length decreased over time as a result of bone apposition in the Dist group. However, in the FK + Dist group, the bone defect length remained the same.
    Conclusions. These results suggest that bone mass and architecture strongly affect the tolerance to the overloading and adaptation with bone apposition in condylar resorption site.

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  • 新潟大学歯学部における統合型模型実習の取り組み

    秋葉 奈美, 長澤 麻沙子, 小野 和宏, 前田 健康, 魚島 勝美

    日本歯科医学教育学会雑誌   33 ( 2 )   106 - 114   2017年8月

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    記述言語:日本語   出版者・発行元:(一社)日本歯科医学教育学会  

    本邦では、社会の急速な高齢化に伴い、高度な治療が求められる患者や全身的な疾患を有する患者が増加している。また、患者の意識変化によって、より質の高い治療を求められることが多くなっている結果、臨床実習に協力していただける患者が減少している。このような背景から、学生は限られた患者を対象に臨床技能の習得をし、卒業時には相応の知識と臨床能力を確保しなければならない。この問題を解決するための手段として、新潟大学歯学部では1つの模型の中に様々な病態を具備した6種類のオリジナルの模型を開発し、治療計画立案を含めた統合的な模型実習を導入している。実習は臨床実習直前の5年前期に実施され、基礎模型実習と臨床実習をつなぐと共に、不足する臨床実習を補完する役割を果たす。模型には歯石除去必要部位、抜歯、充填、歯内治療、分割抜歯適応歯および歯冠修復、不良補綴歯、欠損補綴適応部位を含む。学生は診断・治療計画立案から歯科処置までをファントムに装着した模型上で実際の臨床をシミュレートする形で行う。現在、本実習の評価はルーブリックを用いて行っているが、評価方法に関しては今後も継続的に検討する必要がある。実習後の学生アンケートからは実習が有意義なものであったとの回答を得ており、知識や技術を使いこなす統合的な能力の育成に有効であることが示唆された。(著者抄録)

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  • 診療参加型歯科臨床実習におけるweb公開型電子ポートフォリオの開発と運用

    小田 陽平, 小野 和宏, 藤井 規孝, 小林 正治, 前田 健康

    日本歯科医学教育学会雑誌   33 ( 2 )   65 - 73   2017年8月

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    記述言語:日本語   出版者・発行元:(一社)日本歯科医学教育学会  

    近年、歯学部・歯科大学では技能教育の改善が求められ、診療参加型臨床実習の実践が推奨されている。本学では以前より、学生が直接歯科医療行為に参加する臨床実習を実施しているが、その学習成果、すなわち、現実的な状況で知識や技術を使いこなせる統合的な能力としての「臨床能力」を、いかにして直接評価すべきか対応に苦慮してきた。また、学生が実際の診療の中でどのようなことを感じ、学習し、どのような指導を受けたか、という学習過程そのものが、必ずしも整理された形で実体化されていないという問題点があった。そこで、臨床能力を評価するためのルーブリックを定め、学生の臨床能力を客観性のある到達レベルとして把握するとともに、日々の臨床実習での学習活動を記載するポートフォリオと組み合わせて運用することにより、臨床実習で行っている教育そのものを経時的に記録・評価し、実体化するという発想にいたった。このような背景から電子ポートフォリオの開発と運用を開始したが、学生・教員の受け入れはスムースであった。臨床実習が進むにつれ、ポートフォリオに記載された教員による学生の評価は向上しており、経験を積むことによる学生の臨床能力向上が反映されたものと推察された。以上のことから、臨床実習における電子ポートフォリオの導入は、学生自身の振り返りや教員の学生指導にとって有用であると考えられた。(著者抄録)

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  • Differential Expression and Roles of Secreted Frizzled-Related Protein 5 and the Wingless Homolog Wnt5a in Periodontitis 査読

    T. Maekawa, P. Kulwattanaporn, K. Hosur, H. Domon, M. Oda, Y. Terao, T. Maeda, G. Hajishengallis

    JOURNAL OF DENTAL RESEARCH   96 ( 5 )   571 - 577   2017年5月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:SAGE PUBLICATIONS INC  

    The Wingless/integrase-1 (Wnt) family of protein ligands and their functional antagonists, secreted frizzled-related proteins (sFRPs), regulate various biological processes ranging from embryonic development to immunity and inflammation. Wnt5a and sFRP5 comprise a typical ligand/antagonist pair, and the former molecule was recently detected at the messenger RNA (mRNA) level in human periodontitis. The main objective of this study was to investigate the interrelationship of expression of Wnt5a and sFRP5 in human periodontitis (as compared to health) and to determine their roles in inflammation and bone loss in an animal model. We detected both Wnt5a and sFRP5 mRNA in human gingiva, with Wnt5a dominating in diseased and sFRP5 in healthy tissue. Wnt5a and sFRP5 protein colocalized in the gingival epithelium, suggesting epithelial cell expression, which was confirmed in cultured human gingival epithelial cells (HGECs). The HGEC expression of Wnt5a and sFRP5 was differentially regulated by a proinflammatory stimulus (lipopolysaccharide [LPS] from Porphyromonas gingivalis) in a manner consistent with the clinical observations (i.e., LPS upregulated Wnt5a and downregulated sFRP5). In HGECs, exogenously added Wnt5a enhanced whereas sFRP5 inhibited LPS-induced inflammation, as monitored by interleukin 8 production. Consistent with this, local treatment with sFRP5 in mice subjected to ligature-induced periodontitis inhibited inflammation and bone loss, correlating with decreased numbers of osteoclasts in bone tissue sections. As in humans, mouse periodontitis was associated with high expression of Wnt5a and low expression of sFRP5, although this profile was reversed after treatment with sFRP5. In conclusion, we demonstrated a novel reciprocal relationship between sFRP5 and Wnt5a expression in periodontal health and disease, paving the way to clinical investigation of the possibility of using the Wnt5a/sFRP5 ratio as a periodontitis biomarker. Moreover, we showed that sFRP5 blocks experimental periodontal inflammation and bone loss, suggesting a promising platform for the development of a new host modulation therapy in periodontitis.

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  • 新潟大学歯学部臨床実習における臨床能力評価

    藤井 規孝, 竹中 彰治, 多部田 康一, 佐藤 直子, 秋葉 奈美, 小田 陽平, 勝見 祐二, 小野 和宏, 前田 健康

    日本歯科医学教育学会雑誌   33 ( 1 )   4 - 11   2017年4月

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    記述言語:日本語   出版者・発行元:(一社)日本歯科医学教育学会  

    歯学教育を改善するために、各大学歯学部・歯科大学においてさまざまな取り組みが行われている。なかでも、診療参加型の臨床実習を充実させることは重視されており、それぞれの歯学部・歯科大学は信頼される歯科医師を輩出するという最も大きな社会的命題を果たすべく努力を続けている。新潟大学歯学部では、卒業生の質の保証を目的に、臨床実習においてACCEPT Projectを立ち上げ、「ACKPIS」と称する方法を用いて学生の臨床能力を評価してきた。ACKPISは診療を自験する学生に対し、どのような専門領域においても必要不可欠となる6つの基本項目を、それぞれの専門処置を評価課題として確認するものである。今回、ACKPISの効果を検証するために、平成28年度の受検生にアンケート調査を行った。ACKPISの基本項目は、ADEAやGDCが提唱する歯科医師に求められるコンピテンシーの中にも該当する記述がみられ、国内外で医師に対して行われている臨床能力評価法同様、診療現場でのフィードバックを含んでいることから、歯学生の臨床能力を評価するために適当であると考えられた。また、アンケート結果から、受検した学生もACKPISの必要性や重要性を認識し、受検方法や合否判定の妥当性を認めていることが明らかになった。以上のことから、ACKPISは臨床実習中の学生の臨床能力を評価するための有用な方法となり得ることが示唆された。(著者抄録)

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  • Regional regulation of Filiform tongue papillae development by Ikkα/Irf6. 査読

    Kawasaki M, Kawasaki K, Oommen S, Blackburn J, Watanabe M, Nagai T, Kitamura A, Maeda T, Liu B, Schmidt-Ullrich R, Akiyama T, Inoue J, Hammond NL, Sharpe PT, Ohazama A

    Developmental dynamics : an official publication of the American Association of Anatomists   245 ( 9 )   937 - 946   2016年9月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

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  • Regional Regulation of Filiform Tongue Papillae Development by Ikk alpha/Irf6 査読

    Maiko Kawasaki, Katsushige Kawasaki, Shelly Oommen, James Blackburn, Momoko Watanabe, Takahiro Nagai, Atsushi Kitamura, Takeyasu Maeda, Bigang Liu, Ruth Schmidt-Ullrich, Taishin Akiyama, Jun-Ichiro Inoue, Nigel L. Hammond, Paul T. Sharpe, Atsushi Ohazama

    DEVELOPMENTAL DYNAMICS   245 ( 9 )   937 - 946   2016年9月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:WILEY  

    Background: Non-gustatory filiform papillae play critical roles in helping to grip food, drawing food to the esophagus, cleaning the mouth, and spreading saliva. The molecular mechanisms of filiform tongue papillae development however are not fully understood. Results: We found Ikk alpha and Irf6 expression in developing tongue epithelium, and describe here specific tongue abnormalities in mice with mutation of these genes, indicating a role for Ikk alpha and Irf6 in filiform papillae development. Ikk alpha and Irf6 mutant tongues showed ectopic vertical epithelium at the midline, while lateral sides of mutant tongues adhered to the oral mucosa. Both the ectopic median vertical epithelium and adhered epithelium exhibited the presence of filiform tongue papillae, whereas epithelium between the median vertical epithelium and adhered tongue showed a loss of filiform tongue papillae. Timing of filiform papillae development was found to be slightly different between the midline and lateral regions of the wild-type tongue. Conclusions: Filiform papillae thus develop through distinct molecular mechanisms between the regions of tongue dorsum in the medio-lateral axis, with some filiform papillae developing under the control of Ikk alpha and Irf6. Developmental Dynamics 245: 937-946, 2016. (C) 2016 Wiley Periodicals, Inc.

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  • Spatio-temporal expression of Sox genes in murine palatogenesis 査読

    Momoko Watanabe, Katsushige Kawasaki, Maiko Kawasaki, Thantrira Portaveetus, Shelly Oommen, James Blackburn, Takahiro Nagai, Atsushi Kitamura, Atsushi Nishikawa, Yasumitsu Kodama, Ritsuo Takagi, Takeyasu Maeda, Paul T. Sharpe, Atsushi Ohazama

    GENE EXPRESSION PATTERNS   21 ( 2 )   111 - 118   2016年7月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:ELSEVIER SCIENCE BV  

    Members of the Sox gene family play critical roles in many biological processes including organogenesis. We carried out comparative in situ hybridisation analysis of seventeen Sox genes (Sox1-14, 17, 18 and 21) during murine palatogenesis from initiation to fusion of the palatal shelves above the dorsal side of the tongue. At palatal shelf initiation (E12.5), the localized expression of six Sox genes (Sox2, 5, 6, 9,12 and 13) was observed in the shelves, whereas Sox4 and Sox11 showed ubiquitious expression. During the down growth of palatal shelves (E13.5), Sox4, Sox5, and Sox9 exhibited restricted expression to the interior side of the palatal shelves facing the tongue. Following elevation of the palatal shelves (E14.5), Sox2, Sox11 and Sox21 expression was present in the midline epithelial seam. We thus identify dynamic spatio-temporal expression of Sox gene family during the process of palatogenesis. (C) 2016 Elsevier B.V. All rights reserved.

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  • Effects of C-xylopyranoside derivative on epithelial regeneration in an in vitro 3D oral mucosa model 査読

    Atsushi Uenoyama, Ikuko Kakizaki, Aki Shiomi, Naoaki Saito, Yuko Hara, Taro Saito, Hisashi Ohnuki, Hiroko Kato, Ritsuo Takagi, Takeyasu Maeda, Kenji Izumi

    BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY   80 ( 7 )   1344 - 1355   2016年7月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:TAYLOR & FRANCIS LTD  

    Identifying substandard tissue-engineered oral mucosa grafts with a poor epithelium before clinical use is critical to ensure quality assurance/control in regenerative medicine, leading to success of grafting. This study investigated the effects of one of the C-xylopyranoside derivatives, -D-xylopyranoside-n-propane-2-one (XPP), on oral epithelial regeneration. Using a three-dimensional oral mucosa model, we analyzed changes of the epithelial structure, glycosaminoglycan (GAG) synthesis, the expression levels of basement membrane zone markers, and substrates of Akt/mTOR signaling. Compared with the control, 2mM XPP treatment increased the mean and minimal epithelial thickness, and reduced the variation of epithelial thickness. It also stimulated expressions of decorin and syndecan-1 with change of GAG amount and/or composition, and enhanced the expressions of integrin 6, CD44, and Akt/mTOR signaling substrates. These findings suggest that XPP supplementation contributes to consistent epithelial regeneration. Moreover, upregulation of those markers may play a role in increasing the quality of the oral mucosal epithelium.

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  • Contribution of synovial lining cells to synovial vascularization of the rat temporomandibular joint 査読

    Kayoko Nozawa-Inoue, Fumiko Harada, Jin Magara, Atsushi Ohazama, Takeyasu Maeda

    JOURNAL OF ANATOMY   228 ( 3 )   520 - 529   2016年3月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:WILEY-BLACKWELL  

    The lining layer of the synovial membrane in the temporomandibular joint (TMJ) contains two types of lining cells: macrophage-like type A and fibroblast-like type B cells. The type B cells are particularly heterogeneous in their morphology and immunoreactivity, so that details of their functions remain unclear. Some of the type B cells exhibit certain resemblances in their ultrastructure to those of an activated capillary pericyte at the initial stage of the angiogenesis. The articular surface, composed of cartilage and the disc in the TMJ, has few vasculatures, whereas the synovial lining layer is richly equipped with blood capillaries to produce the constituent of synovial fluid. The present study investigated at both the light and electron microscopic levels the immunocytochemical characteristics of the synovial lining cells in the adult rat TMJ, focusing on their contribution to the synovial vascularization. It also employed an intravascular perfusion with Lycopersicon esculentum (tomato) lectin to identify functional vessels in vivo. Results showed that several type B cells expressed desmin, a muscle-specific intermediate filament which is known as the earliest protein to appear during myogenesis as well as being a marker for the immature capillary pericyte. These desmin-positive type B cells showed immunoreactions for vimentin and pericyte markers (neuron-glial 2; NG2 and PDGFRb) but not for the other markers of myogenic cells (MyoD and myogenin) or a contractile apparatus (aSMA and caldesmon). Immunoreactivity for RECA-1, an endothelial marker, was observed in the macrophage-like type A cells. The arterioles and venules inside the synovial folds extended numerous capillaries with RECA-1-positive endothelial cells and desmin-positive pericytes to distribute densely in the lining layer. The distal portion of these capillaries showing RECA-1-immunoreactivity lacked lectin-staining, indicating a loss of blood-circulation due to sprouting or termination in the lining layer. The desmin-positive type B and RECA-1-positive type A cells attached to this portion of the capillaries. Some capillaries in the lining layer also expressed ninein, a marker for sprouting endothelial cells, called tip cells. Since an activated pericyte, macrophage and tip cell are known to act together at the forefront of the vessel sprout during angiogenesis, the desmin-positive type B cell and RECA-1-positive type A cell might serve as these angiogenic cells in the synovial lining layer. Tomato lectin perfusion following decalcification would be a highly useful tool for research on the vasculature of the mineralized tissue. Use of this technique combined with immunohistochemistry should permit future extensive investigations on the presence of the physiological angiogenesis and on the function of the lining cells in the synovial membrane. Key words: angiogenesis; synovial lining cell; synovial membrane; temporomandibular joint; tomato lectin.

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  • Peripheral brain-derived neurotrophic factor modulates regeneration following inferior alveolar nerve injury in rats 査読

    Hiroyuki Yoshikawa, Yurie Yamada, Masayuki Kurose, Kensuke Yamamura, Takeyasu Maeda, Kenji Seo

    Journal of Oral and Facial Pain and Headache   30 ( 4 )   346 - 354   2016年

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:Quintessence Publishing Co. Inc.  

    Aims: To examine the effects of local brain-derived neurotrophic factor (BDNF) produced after nerve injury on the functional regeneration of the damaged nerve. Methods: The inferior alveolar nerve was transected in adult male rats and 1 μg or 10 μg of BDNF antibody was administered at the injury site
    a third group of rats received saline and a fourth group underwent nerve ligation. BDNF mRNA was quantified in the transected tissue and trigeminal ganglion by using realtime polymerase chain reaction (PCR). Head withdrawal thresholds following mechanical (tactile) stimulation (with von Frey filaments) of the mental region were measured for 3 weeks postoperatively. Electromyographic activity of the jaw opening reflex (JOR) was recorded from the anterior belly of the digastric muscle. Results: Within 24 hours, transection induced significant elevation of BDNF mRNA expression in the injured tissue (unpaired t test, P &lt
    .01). The head withdrawal threshold to mechanical stimulation increased at 1 day after transection and then decreased (two-way repeated measures analysis of variance [ANOVA], P &lt
    .001). At 2 weeks after surgery, the head withdrawal threshold was higher than before surgery in the group that received a higher dose of BDNF antibody (ANOVA, P &lt
    .001), but not in the group that received a smaller dose (ANOVA, P &gt
    .05). No significant differences were observed in the latency or threshold of the JOR between saline- and antibody-treated rats (unpaired t test, P &gt
    .05). Conclusion: These results suggest that locally administered BDNF antibody neutralizes nerve injury-induced BDNF at the injury site and thus influences sensorimotor recovery.

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  • Cyclic mechanical pressure-loading alters epithelial homeostasis in a three-dimensional in vitro oral mucosa model: clinical implications for denture-wearers 査読

    A. Shiomi, K. Izumi, A. Uenoyama, T. Saito, N. Saito, H. Ohnuki, H. Kato, M. Kanatani, S. Nomura, H. Egusa, T. Maeda

    JOURNAL OF ORAL REHABILITATION   42 ( 3 )   192 - 201   2015年3月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:WILEY-BLACKWELL  

    Denture-wearing affects the quality and quantity of epithelial cells in the underlying healthy oral mucosa. The physiologic mechanisms, however, are poorly understood. This study aimed to compare histologic changes and cellular responses of an epithelial cell layer to cyclic mechanical pressure-loading mimicking denture-wearing using an organotypic culture system to develop a three-dimensional in vitro oral mucosa model (3DOMM). Primary human oral keratinocytes and fibroblasts were serially grown in a monolayer culture, and cell viability was measured under continuous cyclic mechanical pressure (50kPa) for 7days (cycles of 60min on, 20s off to degas and inject air). Upon initiation of an air-liquid interface culture for epithelial stratification, the cyclic pressure, set to the mode above mentioned, was applied to the 3DOMMs for 7days. Paraffin-embedded 3DOMMs were examined histologically and immunohistochemically. In the monolayer culture, the pressure did not affect the viability of oral keratinocytes or fibroblasts. Few histologic changes were observed in the epithelial layer of the control and pressure-loaded 3DOMMs. Immunohistochemical examination, however, revealed a significant decrease in Ki-67 labelling and an increase in filaggrin and involucrin expression in the suprabasal layer of the pressure-loaded 3DOMMs. Pressure-loading attenuated integrin 1 expression and increased matrix metalloproteinase-9 activity. Incomplete deposition of laminin and type IV collagen beneath the basal cells was observed only in the pressure-loaded 3DOMM. Cyclic pressure-loading appeared to disrupt multiple functions of the basal cells in the 3DOMM, resulting in a predisposition towards terminal differentiation. Thus, denture-wearing could compromise oral epithelial homeostasis.

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  • Excess NF-κB induces ectopic odontogenesis in embryonic incisor epithelium. 査読

    Blackburn J, Kawasaki K, Porntaveetus T, Kawasaki M, Otsuka-Tanaka Y, Miake Y, Ota MS, Watanabe M, Hishinuma M, Nomoto T, Oommen S, Ghafoor S, Harada F, Nozawa-Inoue K, Maeda T, Peterková R, Lesot H, Inoue J, Akiyama T, Schmidt-Ullrich R, Liu B, Hu Y, Page A, Ramírez Á, Sharpe PT, Ohazama A

    Journal of dental research   94 ( 1 )   121 - 128   2015年1月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    DOI: 10.1177/0022034514556707

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  • Expression of Sox genes in tooth development 査読

    Katsushige Kawasaki, Maiko Kawasaki, Momoko Watanabe, Erik Idrus, Takahiro Nagai, Shelly Oommen, Takeyasu Maeda, Nobuko Hagiwara, Jianwen Que, Paul T. Sharpe, Atsushi Ohazama

    INTERNATIONAL JOURNAL OF DEVELOPMENTAL BIOLOGY   59 ( 10-12 )   471 - 478   2015年

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:UNIV BASQUE COUNTRY UPV-EHU PRESS  

    Members of the Sox gene family play roles in many biological processes including organogenesis. We carried out comparative in situ hybridization analysis of seventeen sox genes (Sox 1-14, 17, 18, 21) during murine odontogenesis from the epithelial thickening to the cytodifferentiation stages. Localized expression of five Sox genes (Sox6, 9, 13, 14 and 21) was observed in tooth bud epithelium. Sox13 showed restricted expression in the primary enamel knots. At the early bell stage, three Sox genes (Sox8, 11, 17 and 21) were expressed in pre-ameloblasts, whereas two others (Sox5 and 18) showed expression in odontoblasts. Sox genes thus showed a dynamic spatio-temporal expression during tooth development.

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  • A histologic study of deformation of the mandibular condyle caused by distraction in a rat model 査読

    Naoko Sakagami, Tadaharu Kobayashi, Kayoko Nozawa-Inoue, Kimimitsu Oda, Taku Kojima, Takeyasu Maeda, Chikara Saito

    ORAL SURGERY ORAL MEDICINE ORAL PATHOLOGY ORAL RADIOLOGY   118 ( 3 )   284 - 294   2014年9月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:ELSEVIER SCIENCE INC  

    Objective. This study investigated the bone resorption process of the rat mandibular condyle after mandibular distraction.
    Study Design. Male Wistar rats at 10 weeks of age underwent unilateral mandibular distraction at 0.175 mm per 12 hours for 10 days. Histologic and histochemical analyses were performed at postoperative day 1 and weeks 1 and 3.
    Results. High-resolution computed tomography (micro-CT) observations showed that deformation of the condyle occurred in the anterior region, where a discontinuity of the condylar cartilage layer was found in histologic sections. This destroyed area gathered many osteoclasts. In the central region, disorganization with a thin hypertrophic cell layer was recognizable by day 1 but later thickened. Morphologic recovery of the mandibular condyle could be attained by week 3 in this animal model.
    Conclusions. These morphologic findings indicate that rapid deformation of the condyle, with destruction of the cartilage layer and bone resorption, was caused by artificial distraction.

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  • Effect of local application of an antibody against brain-derived neurotrophic factor on neuroma formation after transection of the inferior alveolar nerve in the rat 査読

    Yessenia M. Valverde Guevara, Hiroyuki Yoshikawa, Isao Saito, Takeyasu Maeda, Kenji Seo

    NEUROREPORT   25 ( 13 )   1069 - 1074   2014年9月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:LIPPINCOTT WILLIAMS & WILKINS  

    This study aimed to examine the contributions of brain-derived neurotrophic factor (BDNF) at the injury site toward neuroma formation and nerve regeneration after inferior alveolar nerve transection. Histological analysis confirmed neuroma formation at 2 weeks after complete transection of the inferior alveolar nerve. A local administration of an antibody to BDNF inhibited connective tissue proliferation at the injury site and promoted nerve fiber integrity. Fluorogold labeling showed a significantly higher number of labeled cells in the trigeminal ganglion in the anti-BDNFtreated group compared with the vehicle control group. In-situ hybridization histochemistry showed intense signals for tropomyosin receptor kinase B mRNA in the area of the injury site containing fibrous or granular tissue in the anti-BDNF-treated group. In contrast, these signals were close to the detection limit in the area of the perineurium in intact nerve trunks, indicating that the signals were expressed by fibroblasts within the connective tissue. These findings suggest that antagonization of endogenous BDNF induced by nerve injury reduces neuroma formation, without inhibiting damaged axon regeneration. (C) 2014 Wolters Kluwer Health vertical bar Lippincott Williams & Wilkins.

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  • R-spondins/Lgrs expression in tooth development 査読

    Maiko Kawasaki, Thantrira Porntaveetus, Katsushige Kawasaki, Shelly Oommen, Yoko Otsuka-Tanaka, Mitsue Hishinuma, Takato Nomoto, Takeyasu Maeda, Keiyo Takubo, Toshio Suda, Paul T. Sharpe, Atsushi Ohazama

    DEVELOPMENTAL DYNAMICS   243 ( 6 )   844 - 851   2014年6月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:WILEY-BLACKWELL  

    Background: Tooth development is highly regulated in mammals and it is regulated by networks of signaling pathways (e. g. Tnf, Wnt, Shh, Fgf and Bmp) whose activities are controlled by the balance between ligands, activators, inhibitors and receptors. The members of the R-spondin family are known as activators of Wnt signaling, and Lgr4, Lgr5, and Lgr6 have been identified as receptors for R-spondins. The role of R-spondin/Lgr signaling in tooth development, however, remains unclear. Results: We first carried out comparative in situ hybridization analysis of R-spondins and Lgrs, and identified their dynamic spatio-temporal expression in murine odontogenesis. R-spondin2 expression was found both in tooth germs and the tooth-less region, the diastema. We further examined tooth development in R-spondin2 mutant mice, and although molars and incisors exhibited no significant abnormalities, supernumerary teeth were observed in the diastema. Conclusions: R-spondin/Lgr signaling is thus involved in tooth development. Developmental Dynamics 243:844-851, 2014. (c) 2014 Wiley Periodicals, Inc.

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  • Zoledronic acid impairs re-epithelialization through down-regulation of integrin αvβ6 and transforming growth factor beta signalling in a three-dimensional in vitro wound healing model. 査読

    Saito T, Izumi K, Shiomi A, Uenoyama A, Ohnuki H, Kato H, Terada M, Nozawa-Inoue K, Kawano Y, Takagi R, Maeda T

    International journal of oral and maxillofacial surgery   43 ( 3 )   373 - 380   2014年3月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

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  • 大学学習法へのパフォーマンス評価導入における実践的課題

    小野 和宏, 西山 秀昌, 八木 稔, ステガロユ ロクサーナ, 重谷 佳見, 山村 健介, 井上 誠, 前田 健康, Ono Kazuhiro, Nishiyama Hideyoshi, Yagi Minoru, Stegaroiu Roxana, Shigetani Yoshimi, Yamamura Kensuke, Inoue Makoto, Maeda Takeyasu

    新潟大学高等教育研究   1 ( 2 )   5 - 8   2014年

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    記述言語:日本語   出版者・発行元:新潟大学教育・学生支援機構  

    新潟大学歯学部では,初年次教育として大学学習法を実施している.この授業は講義・演習での知識や技能をもとに,レポート作成とプレゼンテーションを行うもので,その学習活動を通して,間題解決能力,論理的思考力,表現力の開発をねらいとしている.今回,学習成果の直接評価に向けて,パフォーマンス評価を導入すべく,レポートからこれら能力を読み解くルーブリックづくりに取り組んだ.また,この経験からみえてきたパフォーマンス評価の難点とその対処について検討した.その結果,パフォーマンス評価により能力を把握できる可能性が示されたが,導入には教員の理解と協力,能力と関連した適切な評価観点の設定,評価の信頼性を高める工夫,評価と指導の一体化を考慮した授業デザインなど,いくつかの課題があることも明らかになった.The curriculum of Niigata University Faculty of Dentistry provides the &quot;Study Skills&quot; course as first year education. The ultimate goal of this course is to develop student abilities of problem solving, logical thinking and expression. For that purpose, each student is guided through the process of writing a report and presenting its content. To access the degree to which these abilities were achieved, we created a rubric and used it to evaluate the student reports. Based on the experience of this trial, we discussed problems for the introduction of performance assessment into the course. Although performance assessment has a high potential to grasp these abilities, there are some practical problems that may interfere with its introduction, such as &quot;how to involve the faculty in assessment&quot;, &quot;how to establish dimensions of the rubric that appropriately relate with the student abilities&quot;, &quot;how to increase the reliability of assessment&quot;, &quot;how to design the course to allow for a connection between assessment and instruction&quot;.

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  • Distinct expression patterns and roles of aldehyde dehydrogenases in normal oral mucosa keratinocytes: differential inhibitory effects of a pharmacological inhibitor and RNAi-mediated knockdown on cellular phenotype and epithelial morphology 査読

    Hiroko Kato, Kenji Izumi, Taro Saito, Hisashi Ohnuki, Michiko Terada, Yoshiro Kawano, Kayoko Nozawa-Inoue, Chikara Saito, Takeyasu Maeda

    HISTOCHEMISTRY AND CELL BIOLOGY   139 ( 6 )   847 - 862   2013年6月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:SPRINGER  

    Aldehyde dehydrogenases (ALDHs), enzymes responsible for detoxification and retinoic acid biosynthesis, are considered a potent functional stem cell marker of normal and malignant cells in many tissues. To date, however, there are no available data on ALDH distributions and functions in oral mucosa. This study aims to clarify the levels and types of ALDH expression using immunohistochemistry with accompanying mRNA expression as well as an ALDEFLUOR assay, and to assess phenotypic and histological changes after manipulation of the ALDH activity of oral keratinocytes to increase the potency of a tissue-engineered oral mucosa by a specific ALDH inhibitor, diethylaminobenzaldehyde (DEAB), together with small interfering RNA of ALDH1A3 and ALDH3A1. Results showed the mRNA and cytoplasmic protein expression of ALDH1A3 and ALDH3A1 to be mostly localized in the upper suprabasal layer although no ALDH1A1 immunoreaction was detected throughout the epithelium. Oral keratinocytes with high ALDH activity exhibited a profile of differentiating cells. By pharmacological inhibition, the phenotypic analysis revealed the proliferating cell-population shifting to a more quiescent state compared with untreated cells. Furthermore, a well-structured epithelial layer showing a normal differentiation pattern and a decrease in Ki-67 immunopositive basal cells was developed by DEAB incubation, suggesting a slower turnover rate efficient to maintain undifferentiated cells. Histological findings of a regenerated oral epithelium by ALDH1A3 siRNA were similar to those when treated with DEAB while ALDH3A1 siRNA eradicated the epithelial regenerative capacity. These observations suggest the effects of phenotypic and morphological alterations by DEAB on oral keratinocytes are mainly consequent to the inhibition of ALDH1A3 activity.

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  • Histochemical aspects of the vascular invasion at the erosion zone of the epiphyseal cartilage in MMP-9-deficient mice 査読

    Taku Kojima, Tomoka Hasegawa, Paulo Henrique Luiz de Freitas, Tomomaya Yamamoto, Muneteru Sasaki, Keisuke Horiuchi, Hiromi Hongo, Tamaki Yamada, Naoko Sakagami, Naoaki Saito, Michiko Yoshizawa, Tadaharu Kobayashi, Takeyasu Maeda, Chikara Saito, Norio Amizuka

    BIOMEDICAL RESEARCH-TOKYO   34 ( 3 )   119 - 128   2013年6月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:BIOMEDICAL RESEARCH PRESS LTD  

    We have histologically examined vascular invasion and calcification of the hypertrophic zone during endochondral ossification in matrix metalloproteinase (MMP)-9 deficient (MMP-9(-/-)) mice and in their littermates at 3 days, 3 weeks and 6 weeks after birth. Capillaries and osteoclasts at the chondro-osseous junction showed an intense MMP-9 immunopositivity, suggesting that they recognize chemical properties of cartilaginous matrices, and then release MMP-9 for cartilage degradation. CD31-positive capillaries and tartrate-resistant acid phosphatase-reactive osteoclasts could be found in the close proximity in the region of chondro-osseous junction in MMP-9(-/-) mice, while in wild-type mice, vascular invasion preceded osteoclastic migration into the epiphyseal cartilage. Although MNP-9(-/-) mice revealed larger hypertrophic zones, the index of calcified area was significantly smaller in MMP-9(-/-) mice. Interestingly, the lower layer of the MMP-9(-/-) hypertrophic zone showed intense MMP-13 staining, which could not be observed in wild-type mice. This indicates that MMP-13 may compensate for MMP-9 deficiency at that specific region, but not to a point at which the deficiency could be completely rescued. In conclusion, it seems that MMP-9 is the optimal enzyme for cartilage degradation during endochondral ossification by controlling vascular invasion and subsequent osteoclastic migration.

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  • [Results of a questionnaire on efforts to increase research-oriented doctors]. 査読

    Hiroyuki Yaginuma, George Matsumura, Chisato Mori, Takeyasu Maeda, Nobukazu Araki, Yasuko Noda, Kazunori Nakajima, Mitsuhiro Kawata, Shigeo Okabe

    Kaibogaku zasshi. Journal of anatomy   88 ( 1-2 )   3 - 8   2013年3月

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    記述言語:日本語  

    We surveyed medical and dental schools to promote the exchange of information about university efforts to increase the number of research-oriented doctors. Periods in which students rotate through laboratories to conduct research were reported by more than two thirds of universities. Many comments asserted that these efforts are effective. However, a small number of respondents reported low student motivation and insufficient time for laboratory experience. MD-PhD courses, in which students take a leave of absence in the middle of undergraduate training and follow a PhD curriculum, have been employed by more than 10 universities. However, relatively few students have chosen such programs. Modified MD-PhD courses have recently been introduced by several universities. In these courses, by taking part of the graduate school curriculum in advance, undergraduate students can shorten the time they spend in graduate school. Students who take such courses are increasing. There were many opinions that extra positions and financial support for research-oriented doctors are effective and should be enhanced. There were also many opinions that emphasize the importance of identifying research-oriented students, improving laboratory working environments, attending academic meetings and inter-university consortia to maintain students' motivation, and promoting collaboration with departments of clinical medicine.

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  • Observation of the Bone Surrounding an Overloaded Implant in a Novel Rat Model 査読

    Masako Nagasawa, Ryohei Takano, Takeyasu Maeda, Katsumi Uoshima

    INTERNATIONAL JOURNAL OF ORAL & MAXILLOFACIAL IMPLANTS   28 ( 1 )   109 - 116   2013年1月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:QUINTESSENCE PUBLISHING CO INC  

    Purpose: Although the recent success rates of dental implants are quite high, it is still true that loading during the bone healing period may be more likely to discourage osseointegration and that occlusal overloading might result in implant failure, even after osseointegration is established. The purposes of this study were to establish an appropriate experimental animal model and to histologically assess degenerative changes in established osseointegration under early and excessive occlusal loading. Materials and Methods: Forty rats were divided into control and experimental groups. The maxillary first and second molars on both sides were extracted, and machined-surface titanium implants were placed. In the experimental group, 2 or 4 weeks after implant placement, abutments that were designed to overload the implants were attached. Control group implants did not receive abutments and remained in situ 2 or 4 weeks. Sections were prepared and observed histologically. Results: Attrition of occluding opposite teeth and shiny spots on the abutments indicated that this model was useful for histologic investigation of the remodeling and bone changes around implants. Specimens showed remarkable bone loss and deterioration of osseointegration when overloading began at 2 weeks. Overloading applied after 4 weeks of healing induced active bone resorption in remote areas of the implants after 15 days of occlusion, while bone resorption at the interface was limited. Conclusion: The authors successfully established an implant occlusion model using rats. This model revealed degenerative changes in osseointegration and/or in the bone around implants upon excessive occlusal loading. These results emphasize the risks associated with immediate loading and overloading. This is the first study to reveal the possibility of bone loss around overloaded implants in the absence of infection using a small animal model.

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  • Construction and characterization of a tissue-engineered oral mucosa equivalent based on a chitosan-fish scale collagen composite 査読

    Michiko Terada, Kenji Izumi, Hisashi Ohnuki, Taro Saito, Hiroko Kato, Marie Yamamoto, Yoshiro Kawano, Kayoko Nozawa-Inoue, Haruhiko Kashiwazaki, Toshiyuki Ikoma, Junzo Tanaka, Takeyasu Maeda

    JOURNAL OF BIOMEDICAL MATERIALS RESEARCH PART B-APPLIED BIOMATERIALS   100B ( 7 )   1792 - 1802   2012年10月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:WILEY-BLACKWELL  

    This study was designed to (1) assess the in vitro biocompatibility of a chitosancollagen composite scaffold (C3) constructed by blending commercial chitosan and tilapia scale collagen with oral mucosa keratinocytes, (2) histologically and immunohistochemically characterize an ex vivo-produced oral mucosa equivalent constructed using the C3 (EVPOME-C), and (3) compare EVPOME-C with oral mucosa constructs utilizing AlloDerm (R) (EVPOME-A), BioMend (R) Extend (TM) (EVPOME-B), and native oral mucosa. C3 scaffold had a well-developed fibril network and a sufficiently small porosity to prevent keratinocytes from growing inside the scaffold after cell-seeding. The EVPOME oral mucosa constructs were fabricated in a chemically defined culture system. After culture at an air-liquid interface, EVPOME-C and EVPOME-B had multilayered epithelium with keratinization, while EVPOME-A had a more organized stratified epithelium. Ki-67 and p63 immunolabeled cells in the basal layer of all EVPOMEs suggested a regenerative ability. Compared with native oral mucosa, the keratin 15 and 10/13 expression patterns in all EVPOMEs showed a less-organized differentiation pattern. In contrast to the beta 1-integrin and laminin distribution in EVPOME-A and native oral mucosa, the subcellular deposition in EVPOME-C and EVPOME-B indicated that complete basement membrane formation failed. These findings demonstrated that C3 has a potential application for epithelial tissue engineering and provides a new potential therapeutic device for oral mucosa regenerative medicine. (C) 2012 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 2012.

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  • Application of capsaicin together with changed sodium channel blocker QX-414 produces transient attenuation of neuropathic-like pain behavior and serves as novel approach for detection of injury related changes in functional expression of TRPV1 channels. 査読

    Hossain Md. Zakir, Rahman, Md. Mostafeezur, Akiko Suzuki, Suzuro Hitomi, Ikuko Suzuki, Takeyasu Maeda, Kenji Seo, Yoshiaki Yamada, Kensuke Yamamura, Shaya, Lev, Alexander M. Binshtok, Junichi Kitagawa

    PLOS ONE   7 ( 9 )   e44023   2012年9月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

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  • Expression of TRPV1 Channels after Nerve Injury Provides an Essential Delivery Tool for Neuropathic Pain Attenuation 査読

    Hossain Md Zakir, Rahman Md Mostafeezur, Akiko Suzuki, Suzuro Hitomi, Ikuko Suzuki, Takeyasu Maeda, Kenji Seo, Yoshiaki Yamada, Kensuke Yamamura, Shaya Lev, Alexander M. Binshtok, Koichi Iwata, Junichi Kitagawa

    PLOS ONE   7 ( 9 )   e44023   2012年9月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:PUBLIC LIBRARY SCIENCE  

    Increased expression of the transient receptor potential vanilloid 1 (TRPV1) channels, following nerve injury, may facilitate the entry of QX-314 into nociceptive neurons in order to achieve effective and selective pain relief. In this study we hypothesized that the level of QX-314/capsaicin (QX-CAP) - induced blockade of nocifensive behavior could be used as an indirect in-vivo measurement of functional expression of TRPV1 channels. We used the QX-CAP combination to monitor the functional expression of TRPV1 in regenerated neurons after inferior alveolar nerve (IAN) transection in rats. We evaluated the effect of this combination on pain threshold at different time points after IAN transection by analyzing the escape thresholds to mechanical stimulation of lateral mental skin. At 2 weeks after IAN transection, there was no QX-CAP mediated block of mechanical hyperalgesia, implying that there was no functional expression of TRPV1 channels. These results were confirmed immunohistochemically by staining of regenerated trigeminal ganglion (TG) neurons. This suggests that TRPV1 channel expression is an essential necessity for the QX-CAP mediated blockade. Furthermore, we show that 3 and 4 weeks after IAN transection, application of QX-CAP produced a gradual increase in escape threshold, which paralleled the increased levels of TRPV1 channels that were detected in regenerated TG neurons. Immunohistochemical analysis also revealed that non-myelinated neurons regenerated slowly compared to myelinated neurons following IAN transection. We also show that TRPV1 expression shifted towards myelinated neurons. Our findings suggest that nerve injury modulates the TRPV1 expression pattern in regenerated neurons and that the effectiveness of QX-CAP induced blockade depends on the availability of functional TRPV1 receptors in regenerated neurons. The results of this study also suggest that the QX-CAP based approach can be used as a new behavioral tool to detect dynamic changes in TRPV1 expression, in various pathological conditions.

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  • インプラント周囲骨細胞の免疫組織化学的検索

    羽下 麻衣子[辻村], 網塚 憲生, 前田 健康, 吉江 紀夫

    Journal of Oral Biosciences Supplement   2012   119 - 119   2012年9月

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    記述言語:日本語   出版者・発行元:(一社)歯科基礎医学会  

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  • Immunoexpression of aquaporin-1 in the rat periodontal ligament during experimental tooth movement 査読

    Tomoko Yoshii, Fumiko Harada, Isao Saito, Kayoko Nozawa-Inoue, Yoshiro Kawano, Takeyasu Maeda

    BIOMEDICAL RESEARCH-TOKYO   33 ( 4 )   225 - 233   2012年8月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:BIOMEDICAL RESEARCH PRESS LTD  

    This study examined the immunoexpression pattern of aquaporin-1 (AQP1), first identified as a water channel protein, in the periodontal ligament of rat molars during experimental tooth movement to clarify its role in periodontal responses in an overloaded model by the insertion of a piece of elastic band. In the control group without any treatment, the cementoblasts and osteogenic cells as well as the vascular endothelial cells showed AQP1 immunoreaction. In the experimental group, hyalinized tissue and intensely AQP1 positive amorphous structures which were identified as degenerated endothelial cells by immunoelectron microscopy, occurred at the compression side on Days 1 and 3. AQP1 immunoreaction came to be stronger in the intact endothelial cells around the hyalinized tissue. The hyalinized tissue had almost disappeared by Day 5 when many macrophages reactive to acid phosphatase activity appeared. The periodontal width on Day 7 became almost the same as that in the control group. These findings indicate that the hyalinized tissue and damaged AQP1 positive endothelial cells are phagocytized by macrophages which have temporally migrated, and suggest that the surviving endothelial cells with intense AQP1 reaction are involved in periodontal regeneration by capillary sprouting.

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  • Zoledronic acid induces S-phase arrest via a DNA damage response in normal human oral keratinocytes 査読

    Hisashi Ohnuki, Kenji Izumi, Michiko Terada, Taro Saito, Hiroko Kato, Akiko Suzuki, Yoshiro Kawano, Kayoko Nozawa-Inoue, Ritsuo Takagi, Takeyasu Maeda

    ARCHIVES OF ORAL BIOLOGY   57 ( 7 )   906 - 917   2012年7月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:PERGAMON-ELSEVIER SCIENCE LTD  

    Objective: This study aimed to clarify the effects of zoledronic acid (ZOL) on human primary oral mucosal keratinocytes growing in a monolayer culture and on a tissue-engineered oral mucosal construct.
    Design: Changes in the viability and proliferation of oral keratinocytes incubated with ZOL were measured. Following treatment with 10 mu M ZOL, histological examinations and immunohistochemical analyses for Ki-67, Geminin, and phospho-histone (gamma)-H2A.X were performed on tissue-engineered oral mucosa. Cell cycle distribution and the degree of apoptosis were also measured by flow cytometry. Additionally, we measured the expression of cell cycle regulatory proteins as well as phospho-Chk1 and -Chk2.
    Results: ZOL treatment suppressed cell viability and proliferation in a dose-dependent manner. Compared with untreated tissue-engineered oral mucosa, ZOL treatment resulted in a thinner epithelium in which the basal cells appeared less-organised. This is consistent with the observed significant reduction in the Ki-67 labelling index. In contrast, the Geminin labelling index was significantly higher than that in the untreated sample. In spite of the presence of a few apoptotic cells, ZOL induced an arrest in S-phase, which was confirmed by our observed alterations in the expression levels of cyclin A, B1, p27(KIP1), Rb and phospho-Rb. When the proteasome inhibitor MG132 was added to the ZOL-treated cells, we observed a partial restoration of the expression of cyclin A, cyclin B1, and p27(KIP1). Expression of phospho-Chk1 was detected, and a significant increase in the labelling index of gamma-H2A.X was also seen.
    Conclusions: These results indicate that a 10-mu M ZOL treatment induces a DNA damage response in oral keratinocytes that activates the ubiguitin-mediated proteolysis of cell cycle regulators, resulting in cell cycle arrest and repressive effects on cell viability, proliferation, and epithelial turnover. (C) 2011 Elsevier Ltd. All rights reserved.

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  • Alterations in intermediate filaments expression in disc cells from the rat temporomandibular joint following exposure to continuous compressive force 査読

    Jin Magara, Kayoko Nozawa-Inoue, Akiko Suzuki, Yoshiro Kawano, Kazuhiro Ono, Shuichi Nomura, Takeyasu Maeda

    JOURNAL OF ANATOMY   220 ( 6 )   612 - 621   2012年6月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:WILEY-BLACKWELL  

    The articular disc in the temporomandibular joint (TMJ) that serves in load relief and stabilizing in jaw movements is a dense collagenous tissue consisting of extracellular matrices and disc cells. The various morphological configurations of the disc cells have given us diverse names, such as fibroblasts, chondrocyte-like cells and fibrochondrocytes; however, the characteristics of these cells have remained to be elucidated in detail. The disc cells have been reported to exhibit heterogeneous immunoreaction patterns for intermediate filaments including glial fibrillary acidic protein (GFAP), nestin and vimentin in the adult rat TMJ. Because these intermediate filaments accumulate in the disc cells as tooth eruption proceeds during postnatal development, it might be surmised that the expression of these intermediate filaments in the disc cells closely relates to mechanical stress. The present study was therefore undertaken to examine the effect of a continuous compressive force on the immunoexpression of these intermediate filaments and an additional intermediate filament muscle-specific desmin in the disc cells of the TMJ disc using a rat experimental model. The rats wore an appliance that exerts a continuous compressive load on the TMJ. The experimental period with the appliance was 5 days as determined by previous studies, after which some experimental animals were allowed to survive another 5 days after removal of the appliance. Histological observations demonstrated that the compressive force provoked a remarkable acellular region and a decrease in the thickness of the condylar cartilage of the mandible, and a sparse collagen fiber distribution in the articular disc. The articular disc showed a significant increase in the number of desmin-positive cells as compared with the controls. In contrast, immunopositive cells for GFAP, nestin and vimentin remained unchanged in number as well as intensity. At 5 days after removal of the appliance, both the disc and cartilage exhibited immunohistological and histological features in a recovery process. These findings indicate that the mature articular cells are capable of producing desmin instead of the other intermediate filaments against mechanical stress. The desmin-positive disc cells lacked a-smooth muscle actin (a-SMA) in this study, even though desmin usually co-exists with a-SMA in the vascular smooth muscle cells or pericytes. Because the precursor of a pericyte has such an immunoexpression pattern during angiogenesis, there is a further possibility that the formation of new vessels commenced in response to the extraordinary compressive force.

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  • 骨再生バイオマテリアルと間葉系幹細胞併用による骨再生向上の解析―培養技術を応用した新しい骨再生法の展開― 査読

    小島 拓, 芳澤享子, 小野由起子, 鈴木晶子, 坂上直子, 長谷川智香, 網塚憲生, 織田公光, 前田健康, 齊藤 力

    日本歯科医学会誌   31   34 - 38   2012年3月

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    記述言語:日本語  

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  • 体性感覚研究の進展 歯根膜の感覚受容装置の形態学的基盤―特にルフィニ神経終末について― 査読

    前田健康

    顕微鏡   46 ( 4 )   227 - 232   2011年12月

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    記述言語:日本語   掲載種別:研究論文(学術雑誌)   出版者・発行元:(公社)日本顕微鏡学会  

    歯の支持・固定装置である歯根膜は豊富な知覚神経支配を受けており、咀嚼システムの感覚入力系として機能している。歯根膜の感覚受容器は侵害受容性の自由神経終末と機械受容器にわけられる。歯に加わる刺激は歯根膜機械受容器を介して、さまざまな口腔反射を惹起し、円滑に咀嚼運動を制御している。歯根膜機械受容器として低閾値遅順応性II型の伸展受容器であるルフィニ神経終末が重要である。この神経終末は分枝を繰り返す太い軸索終末と終末シュワン細胞が付随するという特徴をもっている。軸索終末にはカルシウム結合タンパクなどさまざまなタンパクの発現がみられる。歯根膜ルフィニ神経終末の発育・成熟には歯の萠出力や咬合力のような機械刺激の付与が不可欠であり、また高い再生能力を有している。多種の神経栄養因子が時期依存的に作用することによって、歯根膜ルフィニ神経終末の発育・再生が制御されているようである。(著者抄録)

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  • 体性感覚研究の進展 歯根膜の感覚受容装置の形態学的基盤―特にルフィニ神経終末について― 査読

    前田健康

    顕微鏡   46 ( 4 )   227 - 232   2011年12月

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    記述言語:日本語   掲載種別:研究論文(学術雑誌)  

    歯の支持・固定装置である歯根膜は豊富な知覚神経支配を受けており、咀嚼システムの感覚入力系として機能している。歯根膜の感覚受容器は侵害受容性の自由神経終末と機械受容器にわけられる。歯に加わる刺激は歯根膜機械受容器を介して、さまざまな口腔反射を惹起し、円滑に咀嚼運動を制御している。歯根膜機械受容器として低閾値遅順応性II型の伸展受容器であるルフィニ神経終末が重要である。この神経終末は分枝を繰り返す太い軸索終末と終末シュワン細胞が付随するという特徴をもっている。軸索終末にはカルシウム結合タンパクなどさまざまなタンパクの発現がみられる。歯根膜ルフィニ神経終末の発育・成熟には歯の萠出力や咬合力のような機械刺激の付与が不可欠であり、また高い再生能力を有している。多種の神経栄養因子が時期依存的に作用することによって、歯根膜ルフィニ神経終末の発育・再生が制御されているようである。(著者抄録)

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  • Differential roles of CCN family proteins during osteoblast differentiation: Involvement of Smad and MAPK signaling pathways 査読

    Harumi Kawaki, Satoshi Kubota, Akiko Suzuki, Makoto Suzuki, Kumiko Kohsaka, Kenji Hoshi, Toshiya Fujii, Noureddine Lazar, Toshihiro Ohgawara, Takeyasu Maeda, Bernard Perbal, Teruko Takano-Yamamoto, Masaharu Takigawa

    BONE   49 ( 5 )   975 - 989   2011年11月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:ELSEVIER SCIENCE INC  

    CCN family proteins play diverse roles in many aspects of cellular processes such as proliferation, differentiation, adhesion, migration, angiogenesis and survival. In the bone tissue of vertebrate species, the expression of most CCN family members has been observed in osteoblasts. However, their spatial and temporal distributions, as well as their functions, are still only partially understood. In this study, we evaluated the localization of CCN family members in skeletal tissue in vivo and comparatively analyzed the gene expression patterns and functions of the members in murine osteoblasts in primary culture. Immunofluorescent analyses revealed that the CCN family members were differentially produced in osteoblasts and osteocytes. The presence of all Ccn transcripts was confirmed in those osteoblasts. Among the members, CCN1, CCN2, CCN4 and CCN5 were found in osteocytes. CCN4 and CCN5 were distributed in osteocytes located inside of bone matrix as well. Next, we investigated the expression pattern of Ccn family members during osteoblast differentiation. Along with differentiation, most of the members followed proper gene expression patterns: whereas, Ccn4 and Ccn5 showed quite similar patterns. Furthermore, we evaluated the effects of CCN family members on the osteoblastic activities by using recombinant CCN proteins and RNA interference method. Five members of this family displayed positive effects on osteoblast proliferation or differentiation. Of note, CCN3 drastically inhibited the osteoblast activities. Each Ccn specific siRNA could modulate osteoblast activities in a manner expected by the observed effect of respective recombinant CCN protein. In addition, we found that extracellular signal-regulated kinase1/2 and p38 mitogen-activated protein kinase pathways were critically involved in the CCN family member-mediated modification of osteoblast activities.
    Collectively, all Ccn family members were found to be differentially expressed along with differentiation and therefore could participate in progression of the osteoblast lineage. (C) 2011 Elsevier Inc. All rights reserved.

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  • Phenotypes of articular disc cells in the rat temporomandibular joint as demonstrated by immunohistochemistry for nestin and GFAP 査読

    Hitoshi Miyako, Akiko Suzuki, Kayoko Nozawa-Inoue, Jin Magara, Yoshiro Kawano, Kazuhiro Ono, Takeyasu Maeda

    JOURNAL OF ANATOMY   219 ( 4 )   472 - 480   2011年10月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:WILEY-BLACKWELL  

    The articular disc is a dense collagenous tissue containing disc cells that are phenotypically described as chondrocyte-like cells or fibrochondrocytes. Despite the possible existence of these phenotypes in systemic joints, little is known about the detailed classification of the articular disc cells in the temporomandibular joint. In this immunocytochemical study we examined the localization and distribution patterns of nestin and glial fibrillary acidic protein (GFAP) in the articular disc of the rat temporomandibular joint at postnatal day 1, and weeks 1, 2, 4 and 8, based on the status of tooth eruption and occlusion. Nestin and GFAP are intermediate filament proteins whose expression patterns are closely related to cell differentiation and cell migration. Both types of immunopositive cell greatly increased postnatally to a stable level after postnatal week 4, but they showed different distribution patterns and cell morphologies. Nestin-reactive disc cells, which were characterized by a meagre cytoplasm and thin cytoplasmic processes, were scattered in the articular disc, whereas GFAP-positive cells, characterized by broader processes, existed exclusively in the deeper area. In mature discs, the major proportion of articular disc cells exhibited GFAP immunoreactivity. Furthermore, a double-immunostaining demonstrated that the nestin-negative cells, consisting of GFAP-positive and -negative cells, exhibited immunoreactions for heat shock protein 25. These findings indicate that the articular disc cells comprise at least three types in the rat temporomandibular joint and suggest that their expressions closely relate to mechanical loading forces within the joint, including occlusal force, as observed through postnatal development.

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  • ラット顎関節への機械的負荷による関節円板におけるデスミンの発現

    真柄 仁, 野澤 佳世子[井上], 鈴木 晶子, 河野 芳朗, 野村 修一, 前田 健康

    Journal of Oral Biosciences   53 ( Suppl. )   151 - 151   2011年9月

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    記述言語:日本語   出版者・発行元:(一社)歯科基礎医学会  

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  • ラット顎関節滑膜におけるデスミン陽性B型表層細胞

    野澤 佳世子[井上], 鈴木 晶子, 真柄 仁, 河野 芳朗, 寺田 典子, 泉 健次, 前田 健康

    Journal of Oral Biosciences   53 ( Suppl. )   162 - 162   2011年9月

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    記述言語:日本語   出版者・発行元:(一社)歯科基礎医学会  

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  • 初年次教育としての大学学習法の効果と課題

    小野 和宏, 八木 稔, ステガロユ ロクサーナ, 大島 勇人, 西山 秀昌, 八巻 正樹, 前田 健康

    日本歯科医学教育学会雑誌 = Journal of Japanese Association for Dental Education   27 ( 2 )   69 - 77   2011年8月

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  • 初年次教育としての大学学習法の効果と課題

    小野 和宏, 八木 稔, ステガロユ ロクサーナ, 大島 勇人, 西山 秀昌, 八巻 正樹, 前田 健康

    日本歯科医学教育学会雑誌 = Journal of Japanese Association for Dental Education   27 ( 2 )   69 - 77   2011年8月

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  • 新潟大学における初年次教育の役割と課題 査読

    小野和宏, 八木 稔, ステガロユ ロクサーナ, 大島勇人, 西山秀昌, 八巻正樹, 前田健康

    日本歯科医学教育学会雑誌   27 ( 2 )   17 - 25   2011年8月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:日本歯科医学教育学会  

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  • [Final report of the working group for the future planning of the Japanese Association of Anatomists]. 査読

    Hiroyuki Yaginuma, George Matsumura, Chisato Mori, Takeyasu Maeda, Nobukazu Araki, Yasuko Noda, Kazunori Nakajima, Mitsuhiro Kawata, Shigeo Okabe

    Kaibogaku zasshi. Journal of anatomy   86 ( 2 )   39 - 44   2011年6月

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    記述言語:日本語  

    The working group for the future planning of the Japanese Association of Anatomists (JAA) has been working to address the issues that were consulted from the president of JAA since October 2009. After making the interim report in March 2010, a public hearing for general members of the JAA was held and a final report was submitted to the President in January 2011. The report contains the analysis of the current situation, the directions in which we should proceed, and recommendations of concrete actions that JAA should take for each issue. The issues discussed were as follows: 1. Future prospects of anatomy and morphological sciences. How can we maintain the specialties of morphological and anatomical sciences in the rapidly advancing field of life sciences and develop collaborations with other fields? 2. Improvement of the JAA academic meetings. How can we increase JAA members and young participants in the academic meetings of the JAA? 3. Fostering the next generation of young researchers. How can we increase young researchers graduated from the schools of Medicine or Dentistry? 4. Future prospects of education of gross anatomy. Prospects of education in gross anatomy and the body donation registration system in relation with some new cadaver-related movements.

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  • A morphological analysis on the osteocytic lacunar canalicular system in bone surrounding dental implants.

    Haga Maiko, Nozawa-Inoue Kayoko, Li Minqi, Oda Kimimitsu, Yoshie Sumio, Amizuka Norio, Maeda Takeyasu

    Anat Rec (Hoboken)   294 ( 6 )   1074 - 1082   2011年6月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Osseointegration is the most preferable interface of dental implants and newly formed bone. However, the cavity preparation for dental implants often gives rise to empty lacunae or pyknotic osteocytes in bone surrounding the dental implant. This study aimed to examine the chronological alternation of osteocytes in the bone surrounding the titanium implants using a rat model. The distribution of the osteocytic lacunar canalicular system (OLCS) in bone around the titanium implants was examined by silver impregnation according to Bodian&#039;s staining. We also performed double staining for alkaline phosphatase (ALP) and tartrate-resistant acid phosphatase (TRAP), as well as immunohistochemistry for fibroblast growth factor (FGF) 23--a regulator for the serum concentration of phosphorus--and sclerostin, which has been shown to inhibit osteoblastic activities. Newly formed bone and the injured bone at the early stage exhibited an irregularly distributed OLCS and a few osteocytes positive for sclerostin or FGF23, therefore indicating immature bone. Osteocytes in the surrounding bone from Day 20 to Month 2 came to reveal an intense immunoreactivity for sclerostin. Later on, the physiologica

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  • A Morphological Analysis on the Osteocytic Lacunar Canalicular System in Bone Surrounding Dental Implants 査読

    Maiko Haga, Kayoko Nozawa-Inoue, Minqi Li, Kimimitsu Oda, Sumio Yoshie, Norio Amizuka, Takeyasu Maeda

    ANATOMICAL RECORD-ADVANCES IN INTEGRATIVE ANATOMY AND EVOLUTIONARY BIOLOGY   294 ( 6 )   1074 - 1082   2011年6月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:WILEY-BLACKWELL  

    Osseointegration is the most preferable interface of dental implants and newly formed bone. However, the cavity preparation for dental implants often gives rise to empty lacunae or pyknotic osteocytes in bone surrounding the dental implant. This study aimed to examine the chronological alternation of osteocytes in the bone surrounding the titanium implants using a rat model. The distribution of the osteocytic lacunar canalicular system (OLCS) in bone around the titanium implants was examined by silver impregnation according to Bodian&apos;s staining. We also performed double staining for alkaline phosphatase (ALP) and tartrate-resistant acid phosphatase (TRAP), as well as immunohistochemistry for fibroblast growth factor (FGF) 23-a regulator for the serum concentration of phosphorus-and sclerostin, which has been shown to inhibit osteoblastic activities. Newly formed bone and the injured bone at the early stage exhibited an irregularly distributed OLCS and a few osteocytes positive for sclerostin or FGF23, therefore indicating immature bone. Osteocytes in the surrounding bone from Day 20 to Month 2 came to reveal an intense immunoreactivity for sclerostin. Later on, the physiological bone remodeling gradually replaced such immature bone into a compact profile bearing a regularly arranged OLCS. As the bone was remodeled, FGF23 immunoreactivity became more intense, but sclerostin became less so in the well-arranged OLCS. In summary, it seems likely that OLCS in the bone surrounding the dental implants is damaged by cavity formation, but later gradually recovers as bone remodeling takes place, ultimately inducing mature bone. Anat Rec, 294:1074-1082, 2011. (C) 2011 Wiley-Liss, Inc.

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  • 機械的負荷を与えたラット顎関節関節円板におけるデスミンの発現

    真柄 仁, 野澤 佳世子[井上], 鈴木 晶子, 河野 芳朗, 野村 修一, 前田 健康

    新潟歯学会雑誌   41 ( 1 )   52 - 53   2011年6月

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    記述言語:日本語   出版者・発行元:新潟歯学会  

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  • Initial phase of neuropathic pain within a few hours after nerve injury in mice

    Seiji Komagata, Shanlin Chen, Akiko Suzuki, Haruyoshi Yamashita, Ryuichi Hishida, Takeyasu Maeda, Minoru Shibata, Katsuei Shibuki

    Journal of Neuroscience   31 ( 13 )   4896 - 4905   2011年3月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    We tested a hypothesis that the spinal plasticity induced within a few hours after nerve injury may produce changes in cortical activities and an initial phase of neuropathic pain. Somatosensory cortical responses elicited by vibratory stimulation were visualized by transcranial flavoprotein fluorescence imaging in mice. These responses were reduced immediately after cutting the sensory nerves. However, the remaining cortical responses mediated by nearby nerves were potentiated within a few hours after nerve cutting. Nerve injury induces neuropathic pain. In the present study, mice exhibited tactile allodynia 1-2 weeks after nerve injury. Lesioning of the ipsilateral dorsal column, mediating tactile cortical responses, abolished somatic cortical responses to tactile stimuli. However, nontactile cortical responses appeared in response to the same tactile stimuli within a few hours after nerve injury, indicating that tactile allodynia was acutely initiated. Weinvestigated the trigger mechanisms underlying the cortical changes. Endogenous glial cell line-derived neurotrophic factor (GDNF), found in the Meissner corpuscles, induced basal firing_0.1Hzor less in itsA_ tactile afferents, and disruption of the basal firing triggered the potentiation of nontactile cortical responses. Application of 10nMLY341495 [(2S)-2-amino-2-[(1S,2S)-2-carboxycycloprop- 1-yl]-3-(xanth-9-yl) propanoic acid], a specific antagonist of group II metabotropic glutamate receptors (mGluRs), on to the surface of the spinal cord also induced the potentiation of nontactile cortical responses. Together, it is suggested that low-frequency afferent firing produced by GDNF in touch-sensitive nerve fibers continuously activated spinal group II mGluRs and that failure of this activation triggered tactile allodynia. © 2011 the authors.

    DOI: 10.1523/JNEUROSCI.6753-10.2011

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  • Initial Phase of Neuropathic Pain within a Few Hours after Nerve Injury in Mice 査読

    Seiji Komagata, Shanlin Chen, Akiko Suzuki, Haruyoshi Yamashita, Ryuichi Hishida, Takeyasu Maeda, Minoru Shibata, Katsuei Shibuki

    JOURNAL OF NEUROSCIENCE   31 ( 13 )   4896 - 4905   2011年3月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:SOC NEUROSCIENCE  

    We tested a hypothesis that the spinal plasticity induced within a few hours after nerve injury may produce changes in cortical activities and an initial phase of neuropathic pain. Somatosensory cortical responses elicited by vibratory stimulation were visualized by transcranial flavoprotein fluorescence imaging in mice. These responses were reduced immediately after cutting the sensory nerves. However, the remaining cortical responses mediated by nearby nerves were potentiated within a few hours after nerve cutting. Nerve injury induces neuropathic pain. In the present study, mice exhibited tactile allodynia 1-2 weeks after nerve injury. Lesioning of the ipsilateral dorsal column, mediating tactile cortical responses, abolished somatic cortical responses to tactile stimuli. However, nontactile cortical responses appeared in response to the same tactile stimuli within a few hours after nerve injury, indicating that tactile allodynia was acutely initiated. We investigated the trigger mechanisms underlying the cortical changes. Endogenous glial cell line-derived neurotrophic factor ( GDNF), found in the Meissner corpuscles, induced basal firing similar to 0.1 Hz or less in its A beta tactile afferents, and disruption of the basal firing triggered the potentiation of nontactile cortical responses. Application of 10nM LY341495[(2S)-2-amino-2-[(1S, 2S)-2-carboxycyclopropl-yl]-3-(xanth-9-yl)propanoic acid], a specific antagonist of group II metabotropic glutamate receptors (mGluRs), on to the surface of the spinal cord also induced the potentiation of nontactile cortical responses. Together, it is suggested that low-frequency afferent firing produced by GDNF in touch-sensitive nerve fibers continuously activated spinal group II mGluRs and that failure of this activation triggered tactile allodynia.

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  • Detection of acid-sensing ion channel 3 (ASIC3) in periodontal Ruffini endings of mouse incisors 査読

    Farhana Rahman, Fumiko Harada, Isao Saito, Akiko Suzuki, Yoshiro Kawano, Kenji Izumi, Kayoko Nozawa-Inoue, Takeyasu Maeda

    NEUROSCIENCE LETTERS   488 ( 2 )   173 - 177   2011年1月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:ELSEVIER IRELAND LTD  

    The acid-sensing ion channel 3 (ASIC3), a member of the epithelial sodium channel/degenerin (ENaC/DEG) superfamily, has been reported to participate in acid sensing, mechanosensation, and nociception. However, no information is available regarding the precise localization and function of this molecule in the periodontal ligament, which contains abundant sensory nerves originating from the trigeminal ganglion. The present study examined the expression of ASIC3 in the lingual periodontal ligament of mouse incisors by immunohistochemistry. Furthermore, the expression of ASIC3 in the trigeminal ganglion which innervates the periodontal ligament - was investigated at protein (immunohistochemistry and quantitative analysis) and mRNA levels (RT-PCR technique and in situ hybridization histochemistry). Immunohistochemistry for ASIC3 was able to demonstrate dendritic profiles of the periodontal Ruffini endings in the mouse incisors. No thin fibers terminating as nociceptive free nerve endings exhibited ASIC3 immunoreactivity. Double immunofluorescent staining revealed ASIC3 immunoreaction in the axoplasm but not in the ordinary Schwann cells - including the associated terminal Schwann cells. Observation of the trigeminal ganglia showed variously sized neurons expressing ASIC3 immunoreaction; the most intense immunopositivity was found in the small and medium-sized neurons, as confirmed by in situ hybridization histochemistry using a specific cRNA probe. Quantitative analysis on trigeminal ganglion neurons showed that 38.0% of ASIC3 neurons could be categorized as medium-sized neurons which mediate mechanotransduction. These findings suggest that ASIC3 functions as a molecule for mechanosensation in the periodontal Ruffini endings. (C) 2010 Elsevier Ireland Ltd. All rights reserved.

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  • 顎関節関節円板の発達におけるネスチンおよびGFAPの局在変化

    都 仁, 鈴木 晶子, 野澤 佳世子[井上], 真柄 仁, 前田 健康

    新潟歯学会雑誌   40 ( 2 )   193 - 193   2010年12月

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  • 歯科インプラント周囲における骨細胞・骨細管系の組織化学的変化

    羽下 麻衣子, 野澤 佳世子[井上], 李 敏啓, 吉江 紀夫, 網塚 憲生, 前田 健康

    Journal of Oral Biosciences   52 ( Suppl )   128 - 128   2010年9月

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  • 実験的咬合性外傷における歯周組織破壊・改変に伴うアクアポリン1発現の変化

    河野 承子, 河野 芳朗, 三富 智恵, 田口 洋, 早崎 治明, 前田 健康

    Journal of Oral Biosciences   52 ( Suppl )   148 - 148   2010年9月

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  • FGFR3 down-regulates PTH/PTHrP receptor gene expression by mediating JAK/STAT signaling in chondrocytic cell line

    Minqi Li, Yukie Seki, Paulo H.L. Freitas, Masaki Nagata, Taku Kojima, Sara Sultana, Sobhan Ubaidus, Takeyasu Maeda, Junko Shimomura, Janet E. Henderson, Masato Tamura, Kimimitsu Oda, Zhusheng Liu, Ying Guo, Reiko Suzuki, Tsuneyuki Yamamoto, Ritsuo Takagi, Norio Amizuka

    Journal of Electron Microscopy   59 ( 3 )   227 - 236   2010年6月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    The signaling axis comprising the parathyroid hormone (PTH)-related peptide (PTHrP), the PTH/PTHrP receptor and the fibroblast growth factor receptor 3 (FGFR3) plays a central role in chondrocyte proliferation. The Indian hedgehog (IHH) gene is normally expressed in early hypertrophic chondrocytes, and its negative feedback loop was shown to regulate PTH/PTHrP receptor signaling. In this study, we examined the regulation of PTH/PTHrP receptor gene expression in a FGFR3-transfected chondrocytic cell line, CFK2. Expression of IHH could not be verified on these cells, with consequent absence of hypertrophic differentiation. Also, expression of the PTH/PTHrP receptor (75% reduction of total mRNA) and the PTHrP (50% reduction) genes was reduced in CFK2 cells transfected with FGFR3 cDNA. Interestingly, we verified significant reduction in cell growth and increased apoptosis in the transfected cells. STAT1 was detected in the nuclei of the CFK2 cells transfected with FGFR3 cDNA, indicating predominance of the JAK/STAT signaling pathway. The reduction in PTH/PTHrP receptor gene in CFK2 cells overexpressing FGFR3 was partially blocked by treatment with an inhibitor of JAK3 (WHI-P131), but not with an inhibitor of MAPK (SB203580) or JAK2 (AG490). Altogether, these findings suggest that FGFR3 down-regulates PTH/PTHrP receptor gene expression via the JAK/STAT signaling in chondrocytic cells. The Author 2010. Published by Oxford University Press on behalf of Japanese Society of Microscopy. All rights reserved.

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  • FGFR3 down-regulates PTH/PTHrP receptor gene expression by mediating JAK/STAT signaling in chondrocytic cell line 査読

    Minqi Li, Yukie Seki, Paulo H. L. Freitas, Masaki Nagata, Taku Kojima, Sara Sultana, Sobhan Ubaidus, Takeyasu Maeda, Junko Shimomura, Janet E. Henderson, Masato Tamura, Kimimitsu Oda, Zhusheng Liu, Ying Guo, Reiko Suzuki, Tsuneyuki Yamamoto, Ritsuo Takagi, Norio Amizuka

    JOURNAL OF ELECTRON MICROSCOPY   59 ( 3 )   227 - 236   2010年6月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:OXFORD UNIV PRESS  

    The signaling axis comprising the parathyroid hormone (PTH)-related peptide (PTHrP), the PTH/PTHrP receptor and the fibroblast growth factor receptor 3 (FGFR3) plays a central role in chondrocyte proliferation. The Indian hedgehog (IHH) gene is normally expressed in early hypertrophic chondrocytes, and its negative feedback loop was shown to regulate PTH/PTHrP receptor signaling. In this study, we examined the regulation of PTH/PTHrP receptor gene expression in a FGFR3-transfected chondrocytic cell line, CFK2. Expression of IHH could not be verified on these cells, with consequent absence of hypertrophic differentiation. Also, expression of the PTH/PTHrP receptor (75% reduction of total mRNA) and the PTHrP (50% reduction) genes was reduced in CFK2 cells transfected with FGFR3 cDNA. Interestingly, we verified significant reduction in cell growth and increased apoptosis in the transfected cells. STAT1 was detected in the nuclei of the CFK2 cells transfected with FGFR3 cDNA, indicating predominance of the JAK/STAT signaling pathway. The reduction in PTH/PTHrP receptor gene in CFK2 cells overexpressing FGFR3 was partially blocked by treatment with an inhibitor of JAK3 (WHI-P131), but not with an inhibitor of MAPK (SB203580) or JAK2 (AG490). Altogether, these findings suggest that FGFR3 down-regulates PTH/PTHrP receptor gene expression via the JAK/STAT signaling in chondrocytic cells.

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  • Genetic reduction of GABA(A) receptor gamma 2 subunit expression potentiates the immobilizing action of isoflurane 査読

    Kenji Seo, Hiroyuki Seino, Hiroyuki Yoshikawa, Andrey B. Petrenko, Hiroshi Baba, Naoshi Fujiwara, Genji Someya, Yoshiro Kawano, Takeyasu Maeda, Masato Matsuda, Takashi Kanematsu, Masato Hirata

    NEUROSCIENCE LETTERS   472 ( 1 )   1 - 4   2010年3月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:ELSEVIER IRELAND LTD  

    Potentiation of inhibitory gamma-aminobutyric acid subtype A (GABA(A)) receptor function is involved in the mechanisms of anesthetic action. The present study examined the immobilizing action of the volatile anesthetic isoflurane in mice with double knockout (DKO) of phospholipase C-related inactive protein (PRIP)-1 and -2. Both of these proteins play important roles in the expression of GABA(A) receptors containing the gamma 2 subunit on the neuronal cell surface. Immunohistochemistry for GABA(A) receptor subunits demonstrated reduced expression of gamma 2 subunits in the spinal cord of the DKO mice. Immunohistochemistry also revealed up-regulation of the alpha 1 and beta 3 subunits even though there were no apparent differences in the immunoreactivities for the beta 2 subunits between wild-type and DKO mice. The tail-clamp method was used to evaluate the anesthetic/immobilizing effect of isoflurane and the minimum alveolar concentration (MAC) was significantly lower in DKO mice compared with wild-type controls (1.07 +/- 0.01% versus 1.36 +/- 0.04% atm), indicating an increased sensitivity to isoflurane in DKO mice. These immunohistochemical and pharmacological findings suggest that reduced expression of the GABA(A) receptor gamma 2 subunit affects the composition and function of spinal GABA(A) receptors and potentiates the immobilizing action of isoflurane. (C) 2010 Elsevier Ireland Ltd. All rights reserved.

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  • 吸収性プレートによる顎骨の再生

    小島 拓, 網塚 憲生, 鈴木 晶子, 芳澤 享子, 齊藤 力, 前田 健康

    日本歯科医学会誌   29   118 - 118   2010年3月

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    記述言語:日本語   出版者・発行元:日本歯科医学会  

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  • 新潟大学歯学部における教員研修(FD)の概要と今後の方向性 査読

    安島久雄, 魚島勝美, 小野和宏, 藤井規孝, 井上 誠, 山村健介, 齋藤 功, 興地隆史, 高木律男, 前田健康

    日歯教誌   26 ( 3 )   349 - 353   2010年

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  • 新潟大学歯学部歯学科のカリキュラム評価 学習成果に関する卒業生の自己評価と教員評価の一致度による検討 査読

    小野和宏, 八木 稔, 大内章嗣, 魚島勝美, 藤井規孝, 林 孝文, 齋藤 功, 興地隆史, 前田健康

    日歯教誌   26 ( 1 )   49 - 57   2010年

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  • Imaging of somatosensory cortical responses elicited by neuropathic pain in mice 査読

    Katsuei Shibuki, Seiji Komagata, Shanlin Chen, Akiko Suzuki, Haruyoshi Yamashita, Ryuichi Hishida, Takeyasu Maeda, Minoru Shibata

    NEUROSCIENCE RESEARCH   68   E30 - E30   2010年

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    記述言語:英語   出版者・発行元:ELSEVIER IRELAND LTD  

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  • Spinal plasticity converting tactile inputs to pain within a few hours after disruption of GDNF-induced low frequency firing in tactile nerves 査読

    Seiji Komagata, Shanlin Chen, Akiko Suzuki, Haruyoshi Yamashita, Ryuichi Hishida, Takeyasu Maeda, Minoru Shibata, Katsuei Shibuki

    NEUROSCIENCE RESEARCH   68   E156 - E156   2010年

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    記述言語:英語   出版者・発行元:ELSEVIER IRELAND LTD  

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  • FGF23 is mainly synthesized by osteocytes in the regularly distributed osteocytic lacunar canalicular system established after physiological bone remodeling

    Sobhan Ubaidus, Minqi Li, Sara Sultana, Paulo Henrique Luiz De Freitas, Kimimitsu Oda, Takeyasu Maeda, Ritsuo Takagi, Norio Amizuka

    Journal of Electron Microscopy   58 ( 6 )   381 - 392   2009年12月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    This study aimed to evaluate whether the immunolocalization of fibroblast growth factor (FGF) 23 and dentin matrix protein 1 (DMP1) is associated with the spatial regularity of the osteocyte lacunar canalicular system(s) (OLCS). Femora of 12-weeks-old male ICR mice were fixed with 4 paraformaldehyde, decalcified with a 10 EDTA solution and then embedded in paraffin. We have devised a triple staining procedure that combines silver impregnation, alkaline phosphatase (ALPase) immunohistochemistry and tartrate-resistant acid phosphatase (TRAPase) enzyme histochemistry on a single paraffin section. This procedure permitted the visualization of ALPase-positive plump osteoblasts and several TRAPase-positive osteoclasts on those bone matrices featuring irregularly arranged OLCS, and of ALPase-positive bone lining cells on the bone matrix displaying the well-arranged OLCS. As observations proceeded from the metaphysis toward the diaphysis, the endosteal cortical bone displayed narrower bands of calcein labeling, accompanied by increased regularity of the OLCS. This implies that the speed of bone deposition during bone remodeling would affect the regularity of the OLCS. While DMP1 was evenly localized in all regions of the cortical bones, FGF23 was more abundantly localized in osteocytes of cortical bones with regularly arranged OLCS. In cortical bones, the endosteal area featuring regular OLCS exhibited more intense FGF23 immunoreaction when compared to the periosteal region, which tended to display irregular OLCS. In summary, FGF23 appears to be synthesized principally by osteocytes in the regularly distributed OLCS that have been established after bone remodeling.

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  • FGF23 is mainly synthesized by osteocytes in the regularly distributed osteocytic lacunar canalicular system established after physiological bone remodeling 査読

    Sobhan Ubaidus, Minqi Li, Sara Sultana, Paulo Henrique Luiz de Freitas, Kimimitsu Oda, Takeyasu Maeda, Ritsuo Takagi, Norio Amizuka

    JOURNAL OF ELECTRON MICROSCOPY   58 ( 6 )   381 - 392   2009年12月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:OXFORD UNIV PRESS  

    This study aimed to evaluate whether the immunolocalization of fibroblast growth factor (FGF) 23 and dentin matrix protein 1 (DMP1) is associated with the spatial regularity of the osteocyte lacunar canalicular system(s) (OLCS). Femora of 12-weeks-old male ICR mice were fixed with 4% paraformaldehyde, decalcified with a 10% EDTA solution and then embedded in paraffin. We have devised a triple staining procedure that combines silver impregnation, alkaline phosphatase (ALPase) immunohistochemistry and tartrate-resistant acid phosphatase (TRAPase) enzyme histochemistry on a single paraffin section. This procedure permitted the visualization of ALPase-positive plump osteoblasts and several TRAPase-positive osteoclasts on those bone matrices featuring irregularly arranged OLCS, and of ALPase-positive bone lining cells on the bone matrix displaying the well-arranged OLCS. As observations proceeded from the metaphysis toward the diaphysis, the endosteal cortical bone displayed narrower bands of calcein labeling, accompanied by increased regularity of the OLCS. This implies that the speed of bone deposition during bone remodeling would affect the regularity of the OLCS. While DMP1 was evenly localized in all regions of the cortical bones, FGF23 was more abundantly localized in osteocytes of cortical bones with regularly arranged OLCS. In cortical bones, the endosteal area featuring regular OLCS exhibited more intense FGF23 immunoreaction when compared to the periosteal region, which tended to display irregular OLCS. In summary, FGF23 appears to be synthesized principally by osteocytes in the regularly distributed OLCS that have been established after bone remodeling.

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  • Intermittent PTH Administration Stimulates Pre-Osteoblastic Proliferation Without Leading to Enhanced Bone Formation in Osteoclast-Less c-fos(-/-) Mice 査読

    Paulo Henrique Luiz de Freitas, Minqi Li, Tadashi Ninomiya, Midori Nakamura, Sobhan Ubaidus, Kimimitsu Oda, Nobuyuki Udagawa, Takeyasu Maeda, Ritsuo Takagi, Norio Amizuka

    JOURNAL OF BONE AND MINERAL RESEARCH   24 ( 9 )   1586 - 1597   2009年9月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:WILEY-BLACKWELL  

    This study aimed to investigate the behavior and ultrastructure of osteoblastic cells after intermittent PTH treatment and attempted to elucidate the role of osteoclasts on the mediation of PTH-driven bone anabolism. After administering PTH intermittently to wildtype and c-fos(-/-) mice, immunohistochemical, histomorphometrical, ultrastructural, and statistical examinations were performed. Structural and kinetic parameters related to bone formation were increased in PTH-treated wildtype mice, whereas in the osteoclast-deficient c-fos(-/-) mice, there were no significant differences between groups. In wildtype and knockout mice, PTH administration led to significant increases in the number of cells double-positive for alkaline phosphatase and BrdU, suggesting active pre-osteoblastic proliferation. Ultrastructural examinations showed two major pre-osteoblastic subtypes: one rich in endoplasmic reticulum (ER), the hypER cell, and other with fewer and dispersed ER, the misER cell. The latter constituted the most abundant preosteoblastic phenotype after PTH administration in the wildtype mice. In c-fos(-/-) mice, misER cells were present on the bone surfaces but did not seem to be actively producing bone matrix. Several misER cells were shown to be positive for EphB4 and were eventually seen rather close to osteoclasts in the PTH-administered wildtype mice. We concluded that the absence of osteoclasts in c-fos(-/-) mice might hinder PTH-driven bone anabolism and that osteoclastic presence may be necessary for full osteoblastic differentiation and enhanced bone formation seen after intermittent PTH administration. J Bone Miner Res 2009;24:1586-1597. Published online on April 27, 2009; doi: 10.1359/JBMR.090413

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  • Intermittent PTH Administration Stimulates Pre-Osteoblastic Proliferation Without Leading to Enhanced Bone Formation in Osteoclast-Less c-fos(-/-) Mice 査読

    Paulo Henrique Luiz de Freitas, Minqi Li, Tadashi Ninomiya, Midori Nakamura, Sobhan Ubaidus, Kimimitsu Oda, Nobuyuki Udagawa, Takeyasu Maeda, Ritsuo Takagi, Norio Amizuka

    JOURNAL OF BONE AND MINERAL RESEARCH   24 ( 9 )   1586 - 1597   2009年9月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:WILEY-BLACKWELL  

    This study aimed to investigate the behavior and ultrastructure of osteoblastic cells after intermittent PTH treatment and attempted to elucidate the role of osteoclasts on the mediation of PTH-driven bone anabolism. After administering PTH intermittently to wildtype and c-fos(-/-) mice, immunohistochemical, histomorphometrical, ultrastructural, and statistical examinations were performed. Structural and kinetic parameters related to bone formation were increased in PTH-treated wildtype mice, whereas in the osteoclast-deficient c-fos(-/-) mice, there were no significant differences between groups. In wildtype and knockout mice, PTH administration led to significant increases in the number of cells double-positive for alkaline phosphatase and BrdU, suggesting active pre-osteoblastic proliferation. Ultrastructural examinations showed two major pre-osteoblastic subtypes: one rich in endoplasmic reticulum (ER), the hypER cell, and other with fewer and dispersed ER, the misER cell. The latter constituted the most abundant preosteoblastic phenotype after PTH administration in the wildtype mice. In c-fos(-/-) mice, misER cells were present on the bone surfaces but did not seem to be actively producing bone matrix. Several misER cells were shown to be positive for EphB4 and were eventually seen rather close to osteoclasts in the PTH-administered wildtype mice. We concluded that the absence of osteoclasts in c-fos(-/-) mice might hinder PTH-driven bone anabolism and that osteoclastic presence may be necessary for full osteoblastic differentiation and enhanced bone formation seen after intermittent PTH administration. J Bone Miner Res 2009;24:1586-1597. Published online on April 27, 2009; doi: 10.1359/JBMR.090413

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  • GDNF Expression in Terminal Schwann Cells Associated With the Periodontal Ruffini Endings of the Rat Incisors During Nerve Regeneration 査読

    Megumi Ohishi, Fumiko Harada, Farhana Rahman, Isao Saito, Yoshiro Kawano, Kayoko Nozawa-Inoue, Takeyasu Maeda

    ANATOMICAL RECORD-ADVANCES IN INTEGRATIVE ANATOMY AND EVOLUTIONARY BIOLOGY   292 ( 8 )   1182 - 1191   2009年8月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:WILEY  

    The terminal Schwann cells (TSCs) which play crucial roles in regeneration of the periodontal Ruffini endings (RE) exhibit immunoreaction for glial cell line-derived neurotrophic factor (GDNF). However, no information is available regarding the role of GDNF in the periodontal RE during nerve regeneration. This study was undertaken to examine the changes in GDNF expression in the rat periodontal RE following transection of the inferior alveolar nerve (IAN) using immunohistochemistry for GDNF and S-100 protein, a marker for the TSCs. We additionally investigated the changes in expression of GDNF in the trigeminal ganglion (TG) at protein and mRNA levels. A transection to IAN induced a disappearance of the TSCs from the alveolus-related part (ARP), followed by a migration of spindle-shaped cells with S-100 but without GDNF immunoreactions into the tooth-related part (TRP) by postoperative (PO) week 2. At PO week 2, GDNF immunoreacted cellular elements increased in number in the ARP although the spindle-shaped cells without GDNF reaction remained in the TRP After PO week 4, many GDNF-positive TSCs appeared in the ARP though the spindle-shaped cells vanished from the TRP. A real time RT-PCR analysis demonstrated the highest elevation of GDNT mRNA in the TG at PO week 2. These findings suggested the involvement of this molecule in the maturation and maintenance of the periodontal RE during regeneration. Taken together with our previous and current studies, it appears that the regeneration of the periodontal RE is controlled by multiple neurotrophins in a stage-specific manner. Anat Rec, 292:1182-1191, 2009. (C) 2009 Wiley-Liss, Inc.

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  • Influence of heat stress to matrix on bone formation 査読

    Keiko Yoshida, Katsumi Uoshima, Kimimitsu Oda, Takeyasu Maeda

    CLINICAL ORAL IMPLANTS RESEARCH   20 ( 8 )   782 - 790   2009年8月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:WILEY-BLACKWELL  

    Objectives
    It is important to know the etiology of implant failure. It has been reported that heat stress during drilling was one of the causes for failure and the threshold was 47 degrees C. However, clinically, we encounter cases in which overheating does not seem to affect osseointegration eventually. The purpose of this study was to assess histologically the spatio-temporal effect of heat stress on bone formation after overheating the bone matrix.
    Material and methods
    Rat calvarial bone was heated to 37 degrees C, 43 degrees C, 45 degrees C and 48 degrees C for 15 min by a temperature stimulator. Paraffin sections were prepared 1, 3 and 5 weeks after heating and investigated histologically under light microscopy. Hematoxylin and eosin staining, alkaline phosphatase (ALP), osteopontin (OPN), heat shock protein 27 (Hsp27) and heat shock protein 70 (Hsp70) immunohistochemistry and tartrate-resistant acid phosphatase (TRAP) enzyme histochemistry were carried out. The area of dead osteocytes was calculated and statistically analyzed. Apoptotic osteocytes were detected by the terminal deoxynucleotidyltransferase-mediated dUTP nick end-labeling (TUNEL) method.
    Results
    Along with the temperature increase, the area of dead osteocytes increased and regeneration of the periosteal membrane was delayed. Hsps- and TUNEL-positive cells were only seen in the 48 degrees C group. Spatio-temporal changes of TRAP- and ALP-positive cell numbers were observed, while OPN expression was mostly absent. Even after 48 degrees C stimulation, bone formation on the calvarial surface was observed after 5 weeks.
    Conclusions
    Although there was a temperature-dependent delay in bone formation after heat stress, the 48 degrees C heat stress did not obstruct bone formation eventually. This delay was probably caused by slow periosteal membrane regeneration.
    To cite this article:Yoshida K, Uoshima K, Oda K, Maeda T. Influence of heat stress to matrix on bone formation.Clin. Oral Impl. Res. 20, 2009; 782-790.doi: 10.1111/j.1600-0501.2008.01654.x.

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  • Influence of heat stress to matrix on bone formation 査読

    Keiko Yoshida, Katsumi Uoshima, Kimimitsu Oda, Takeyasu Maeda

    CLINICAL ORAL IMPLANTS RESEARCH   20 ( 8 )   782 - 790   2009年8月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:WILEY-BLACKWELL  

    Objectives
    It is important to know the etiology of implant failure. It has been reported that heat stress during drilling was one of the causes for failure and the threshold was 47 degrees C. However, clinically, we encounter cases in which overheating does not seem to affect osseointegration eventually. The purpose of this study was to assess histologically the spatio-temporal effect of heat stress on bone formation after overheating the bone matrix.
    Material and methods
    Rat calvarial bone was heated to 37 degrees C, 43 degrees C, 45 degrees C and 48 degrees C for 15 min by a temperature stimulator. Paraffin sections were prepared 1, 3 and 5 weeks after heating and investigated histologically under light microscopy. Hematoxylin and eosin staining, alkaline phosphatase (ALP), osteopontin (OPN), heat shock protein 27 (Hsp27) and heat shock protein 70 (Hsp70) immunohistochemistry and tartrate-resistant acid phosphatase (TRAP) enzyme histochemistry were carried out. The area of dead osteocytes was calculated and statistically analyzed. Apoptotic osteocytes were detected by the terminal deoxynucleotidyltransferase-mediated dUTP nick end-labeling (TUNEL) method.
    Results
    Along with the temperature increase, the area of dead osteocytes increased and regeneration of the periosteal membrane was delayed. Hsps- and TUNEL-positive cells were only seen in the 48 degrees C group. Spatio-temporal changes of TRAP- and ALP-positive cell numbers were observed, while OPN expression was mostly absent. Even after 48 degrees C stimulation, bone formation on the calvarial surface was observed after 5 weeks.
    Conclusions
    Although there was a temperature-dependent delay in bone formation after heat stress, the 48 degrees C heat stress did not obstruct bone formation eventually. This delay was probably caused by slow periosteal membrane regeneration.
    To cite this article:Yoshida K, Uoshima K, Oda K, Maeda T. Influence of heat stress to matrix on bone formation.Clin. Oral Impl. Res. 20, 2009; 782-790.doi: 10.1111/j.1600-0501.2008.01654.x.

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  • Influence of heat stress to matrix on bone formation.

    Yoshida Keiko, Uoshima Katsumi, Oda Kimimitsu, Maeda Takeyasu

    Clin Oral Implants Res   20 ( 8 )   782 - 790   2009年8月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    OBJECTIVES: It is important to know the etiology of implant failure. It has been reported that heat stress during drilling was one of the causes for failure and the threshold was 47 degrees C. However, clinically, we encounter cases in which overheating does not seem to affect osseointegration eventually. The purpose of this study was to assess histologically the spatio-temporal effect of heat stress on bone formation after overheating the bone matrix. MATERIAL AND METHODS: Rat calvarial bone was heated to 37 degrees C, 43 degrees C, 45 degrees C and 48 degrees C for 15 min by a temperature stimulator. Paraffin sections were prepared 1, 3 and 5 weeks after heating and investigated histologically under light microscopy. Hematoxylin and eosin staining, alkaline phosphatase (ALP), osteopontin (OPN), heat shock protein 27 (Hsp27) and heat shock protein 70 (Hsp70) immunohistochemistry and tartrate-resistant acid phosphatase (TRAP) enzyme histochemistry were carried out. The area of dead osteocytes was calculated and statistically analyzed. Apoptotic osteocytes were detected by the terminal deoxynucleotidyltransferase-mediated dUTP nick end-labeling (TUNEL) method. RESULTS: Along with

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  • 新潟大学歯学部歯学科の新教育課程とその評価 査読

    小野和宏, 八木 稔, 大内章嗣, 魚島勝美, 林 孝文, 齋藤 功, 興地隆史, 前田健康, 山田好秋

    新潟歯誌   39 ( 1 )   29 - 40   2009年6月

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  • Behavioural and histological observations of sensory impairment caused by tight ligation of the trigeminal nerve in mice 査読

    Hiroyuki Seino, Kenji Seo, Takeyasu Maeda, Genji Someya

    JOURNAL OF NEUROSCIENCE METHODS   181 ( 1 )   67 - 72   2009年6月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:ELSEVIER SCIENCE BV  

    Dental treatments sometimes cause sensory impairment, especially in the region innervated by the third division of the trigeminal nerve. The most frequent symptoms are loss of sensation and abnormal sensation. Although most studies have addressed the neuropathic symptom "allodynia" using experimental animal models of the infraorbital nerve, there is little information regarding the sensory impairment that frequently occurs clinically. Therefore, different experimental models are required to clarify the mechanisms of the clinical effects, and previous experimental models have been limited to rats. Here, we report a sensory impairment model in mice whose mechanical touch threshold increased after tight ligation of the mental nerve. Habituation before surgery by mechanical touching of the face enabled us to observe the long-term chronological changes in sensation. The mechanical touch thresholds within the mental nerve region were measured for 70 postoperative (PC) days. Changes in the distribution of substance P (SP) were evaluated by immunohistochemistry to clarify the involvement of axonal flow in the sensory impairment and its recovery. The mechanical touch thresholds transiently increased by PO days 2-3, but decreased to the preoperative levels at around PO day 14. Apparent SP immunoreactivity was recognizable on the medial side to the ligation at PO days 2-3 and disappeared at PO day 7. These behavioural and immunohistochemical changes appeared to exhibit similar time courses, suggesting a possible relationship between them. Therefore, we suggest that our experimental mouse model could represent a new model for clarifying the mechanism of the sensory impairment caused by peripheral nerve injury. (C) 2009 Elsevier B.V. All rights reserved.

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  • Immunohistochemical detection of ENaC beta in the terminal Schwann cells associated with the periodontal Ruffini endings of the rat incisor 査読

    Yasumasa Hitomi, Akiko Suzuki, Yoshiro Kawano, Kayoko Nozawa-Inoue, Makoto Inoue, Takeyasu Maeda

    BIOMEDICAL RESEARCH-TOKYO   30 ( 2 )   113 - 119   2009年4月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:BIOMEDICAL RESEARCH PRESS LTD  

    Epithelial sodium channels (ENaCs) are a subfamily of ion channels within the degenerin/ENaC (DEG/ENaC) superfamily. Previous studies have shown the immunolocalization of ENaC in the neural elements of the Cutaneous mechanoreceptors as well as dorsal root and trigeminal ganglion neurons, indicating the involvement of this molecule in mechanotransduction. The present study examined the expression of ENaC beta, a major component of ENaC protein, in the mechanoreceptive Ruffini endings in the periodontal ligament of the rat incisors by immunohistochemistry. The expression of ENaC beta in the trigeminal ganglion-which innervates the periodontal Ruffini endings-was also investigated at the mRNA and protein levels. Furthermore, double staining and a nerve injury experiment were applied to clarify its detailed localization in the periodontal Ruffini endings. ENaC beta immunoreaction in the trigeminal ganglion was recognizable in the comparatively large neurons which have been considered to mediate mechanotransduction. Immunohistochemistry for ENaC beta demonstrated dendritic ramifications of the Ruffini endings as well as the rounded cells in the periodontal ligament. Double staining with ENaC beta and either PGP9.5 or S-100 protein showed immunoreaction for ENaC beta in both the axonal and glial elements in the periodontal ligament. Some ENaC beta positive cells with rounded profiles were reactive to non-specific cholinesterase activity. Furthermore, a transection of the inferior alveolar nerve failed to eliminate the rounded cells with ENaC beta reaction, indicating that they were the terminal Schwann cells associated with the periodontal Ruffini endings. These findings suggest that ENaC beta is a key mechanotransducing channel in the periodontal Ruffini endings. Probably, the terminal Schwann cells together with the axon terminals regulate mechanotransduction in the periodontal endings.

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  • Differential cell-specific location of Cav-1 and Ca2+-ATPase in terminal Schwann cells and mechanoreceptive Ruffini endings in the periodontal ligament of the rat incisor 査読

    Naoyuki Iizuka, Akiko Suzuki, Kayoko Nozawa-Inoue, Yoshiro Kawano, B. G. T. L. Nandasena, Takashi Okiji, Takeyasu Maeda

    JOURNAL OF ANATOMY   214 ( 2 )   267 - 274   2009年2月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:WILEY-BLACKWELL  

    Caveolae are involved in clathrin-independent endocytosis, transcytosis, signal transduction, and tumor suppression - all of which depend on their main constituent protein caveolin families. The periodontal Ruffini ending has been reported to develop a caveola-like structure on the cell membrane of both the axon terminals and Schwann sheaths, suggesting the existence of an axon-Schwann cell interaction in the periodontal Ruffini endings. However, little information is available concerning the functional significance of these caveolae. The present study was undertaken to examine the immunolocalization of caveolin-1, -3 (Cav-1, Cav-3) and Ca2+-ATPase in the periodontal Ruffini endings of the rat incisor. Decalcified sections of the upper jaws were processed for immunocytochemistry at the levels of light and electron microscopy. Some immunostained sections were treated with histochemistry for nonspecific cholinesterase (nChE) activity. Observations showed the periodontal Ruffini endings were immunopositive for Cav-1, but not Cav-3. Immunoreactive products for Cav-1 were confined to caveola-like structures in the cell membranes of the cytoplasmic extensions and cell bodies of the terminal Schwann cells associated with the periodontal Ruffini endings. However, the axonal membranes of the terminals did not express any Cav-1 immunoreaction. Double staining with Ca2+-ATPase and either protein gene product 9.5 (PGP 9.5) or S-100 protein disclosed the co-localization of immunoreactions in the axonal branches of the periodontal Ruffini endings, but not in the terminal Schwann cells. As Ca2+ plays an important role in mechanotransduction, these characteristic immunolocalizations show Cav-1/Ca2+-ATPase might be involved in the quick elimination of intracellular Ca2+ in mechanotransduction.

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  • Immunohistochemical detection of nestin in the periodontal Ruffini endings of the rat incisor 査読

    Shion Saito, Akiko Suzuki, Kayoko Nozawa-Inoue, Yoshiro Kawano, Masaaki Hoshino, Chikara Saito, Takeyasu Maeda

    NEUROSCIENCE LETTERS   449 ( 3 )   195 - 200   2009年1月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:ELSEVIER IRELAND LTD  

    Nestin is an intermediate filament which was first identified in neuroepithelial stem cells. This expression has also been reported in restricted locations in adults. Previous studies have suggested that the periodontal Ruffini endings remain immature in nature even in adulthood. The present study reports on a characteristic expression of immunoreaction for nestin in the periodontal Ruffini endings during postnatal development. RT-PCR analysis detected nestin mRNA in a reverse transcripted cDNA sample from both the rat trigeminal ganglion and periodontal ligament. The nestin immunoreaction existed in the periodontal ligament at postnatal day 3 (PO 3 days), when many spindle-shaped Schwann cells were positive for nestin immunoreaction. At PO 1 week, when periodontal nerve fibers displayed a dendritic fashion, the round cells came to show the nestin immunoreaction. These immunopositive cells were also reactive for S-100 protein and non-specific cholinesterase, indicating that these cells could be categorized as terminal Schwann cells associated with the periodontal Ruffini endings. Some ordinary Schwann cells also exhibited nestin immunoreaction. From PO 2 to 3 weeks, nestin positive terminal Schwarm cells increased in number in accordance with the postnatal development of the periodontal Ruffini endings, while this immuno-expression pattern remained unchanged. Nestin immunoreaction was also recognizable in the satellite cells - but never in the neurons - in the trigeminat ganglion throughout this observation period. This immuno-expression pattern suggests that nestin serves as an intermediate filament for mechanical stability in the periodontal Ruffini endings against external stimuli. (c) 2008 Elsevier Ireland Ltd. All rights reserved.

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  • Detailed Process of Bone Remodeling After Achievement of Osseointegration in a Rat Implantation Model 査読

    Maiko Haga, Noritaka Fujii, Kayoko Nozawa-Inoue, Shuichi Nomura, Kimimitsu Oda, Katsumi Uoshima, Takeyasu Maeda

    Anatomical Record-Advances in Integrative Anatomy and Evolutionary Biology   292 ( 1 )   38 - 47   2009年1月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:WILEY-LISS  

    Osseointegration is regarded as the most appropriate implant-bone interface in dental implantation. However, damaged bone with empty osteocytic lacunae driven by implant cavity preparation remains even after the completion of osseointegration. Although previous studies have suggested the occurrence of bone remodeling around implants, information on its detailed process is meager. Our study aimed to examine the fate of bone around titanium implants after the establishment of osseointegration on an animal model using the rat maxilla. Titanium implants were inserted into prepared bone cavities of the rat maxilla. Bone formation and maturation processes were evaluated by double staining for alkaline phosphatase and tartrate-resistant acid phosphatase, immunohistochemistry for bone matrix proteins, vital staining with calcein, and elemental mapping with an electron probe microanalyzer. Bone with empty osteocytic lacunae or pyknosis remained between the intact preexisting and newly formed woven bones at post 1 month. It gradually decreased to disappear completely by active bone remodeling with a synchronized coordination of alkaline phosphatase-positive osteoblasts and tartrate-resistant acid phosphatase-reactive osteoclasts at post 3 months, thickening to be replaced by compact bone. Dynamic labeling showed two clear lines in the newly formed bone around the implant through this experimental period. Electron probe microanalyzer analysis demonstrated chronologically increased levels of Ca and P in the newly formed bone identical to those in the surrounding bone at post 2.5 months. These findings indicate that continuous bone remodeling after the achievement of osseointegration causes replacement of the damaged bone by compact bone as well as an improvement in bone quality. Anat Rec,292:38-47, 2009. (c) 2008 Wiley-Liss, Inc.

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  • Vitamin k2, a gamma-carboxylating factor of gla-proteins, normalizes the bone crystal nucleation impaired by Mg-insufficiency 査読

    Norio Amizuka, Minqi Li, Masatoshi Kobayashi, Kuniko Hara, Shoji Akahane, Kiichi Takeuchi, Paulo H. L. Freitas, Hidehiro Ozawa, Takeyasu Maeda, Yasuhiro Akiyama

    HISTOLOGY AND HISTOPATHOLOGY   23 ( 11 )   1353 - 1366   2008年11月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:F HERNANDEZ  

    It has been reported that the Mg-insufficient bone is fragile upon mechanical loading, despite its high bone mineral density, while vitamin K2 (MK-4: menatetrenone) improved the mechanical strength of Mg-insufficient bone. Therefore, we aimed to elucidate the ultrastructural properties of bone in rats with dietary Mg insufficiency with and without MK-4 supplementation. Morphological examinations including histochemistry, transmission electron microscopy, electron probe microanalysis (EPMA) and X-ray diffraction were conducted on the femora and tibiae of 4-week-old Wistar male rats fed with 1) a normal diet (control group, 0.09% Mg), 2) a Mg-insufficient diet (low Mg group, 0.006% Mg), or 3) a Mg-insufficient diet supplemented with MK-4 (MK-4 group, 0.006% Mg, 0.03% MK-4). MK-4 appeared to inhibit the osteoclastic bone resorption that is stimulated by Mg insufficiency. EPMA analysis, however, revealed an increased concentration of Ca paralleling Mg reduction in the low Mg group. Assessment by X-ray diffraction revealed an abundance of a particular synthetic form of hydroxyapatite in the low Mg group, while control bones featured a variety of mineralized crystals. In addition, Mg-deficient bones featured larger mineral crystals, i.e., crystal overgrowth. This crystalline aberration in Mg-insufficient bones induced collagen fibrils to mineralize easily, even in the absence of mineralized nodules, which therefore led to an early collapse of the fibrils. MK-4 prevented premature collagen mineralization by normalizing the association of collagen fibrils with mineralized nodules. Thus, MK-4 appears to rescue the impaired collagen mineralization caused by Mg insufficiency by promoting a re-association of the process of collagen mineralization with mineralized nodules.

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  • Cooperative Regulation of Chondrocyte Differentiation by CCN2 and CCN3 Shown by a Comprehensive Analysis of the CCN Family Proteins in Cartilage 査読

    Harumi Kawaki, Satoshi Kubota, Akiko Suzuki, Noureddine Lazar, Tomohiro Yamada, Tatsushi Matsumura, Toshihiro Ohgawara, Takeyasu Maeda, Bernard Perbal, Karen M. Lyons, Masaharu Takigawa

    JOURNAL OF BONE AND MINERAL RESEARCH   23 ( 11 )   1751 - 1764   2008年11月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:WILEY-BLACKWELL  

    CCN2 is best known as a promoter of chondrocyte differentiation among the CCN family members. and its null mice display skeletal dysmorphisms. However, little is known concerning roles of the other CCN members in chondrocytes. Using both in vivo and in vitro approaches, We conducted a comparative analysis of CCN2-null and wildtype mice to study the roles of CCN2 and the other CCN proteins in cartilage development. Immunohistochemistry was used to evaluate the localization of CCN proteins and other chondrocyte-associated molecules in the two types of mice. Moreover, gene expression levels and the effects of exogenous CCN proteins oil chondrocyte proliferation, differentiation, and the expression of chondrocyte-associated genes in their primary chondrocytes were evaluated. Ccn3 was dramatically upregulated in CCN2-null cartilage and chondrocytes. This upregulation was associated with diminished cell proliferation and delayed differentiation. Consistent with the in vivo findings, CCN2 deletion entirely retarded chondrocyte terminal differentiation and decreased the expression of several chondrocyte-associated genes ill vitro. whereas CcO expression drastically increased. In contrast, the addition Of exogenous CCN2 promoted differentiation strongly and induced the expression of the associated genes. whereas decreasing, the CcO expression. These findings collectively indicate that CCN2 induces chondrocyte differentiation by regulating the expression of chondrocyte-associated genes but that these effects are counteracted by CCN3. The lack of CCN2 caused upregulation of CCN3 in CCN2-null mice, which resulted in the observed phenotypes, Such as the resultant delay of terminal differentiation. The involvement of the PTHrP-Ihh loop in the regulation of CCN3 expression is also suggested.

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  • 生涯学習能力に対するPBLチュートリアルの効果

    小野和宏, 八木稔, 大内章嗣, 魚島勝美, 安島久雄, 林孝文, 齋藤功, 興地隆史, 前田健康

    日本歯科医学教育学会雑誌   24 ( 2 )   145 - 149   2008年8月

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    記述言語:日本語  

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  • Immunocytochemical localization of caveolin-3 in the synoviocytes of the rat temporomandibular joint during development 査読

    Masahiro Niwano, Kayoko Nozawa-Inoue, Akiko Suzuki, Nobuyuki Ikeda, Ritsuo Takagi, Takeyasu Maeda

    ANATOMICAL RECORD-ADVANCES IN INTEGRATIVE ANATOMY AND EVOLUTIONARY BIOLOGY   291 ( 3 )   233 - 241   2008年3月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:WILEY-LISS  

    Caveolins-caveolin-1, -2, -3 (Cav1, 2, 3)-are major components of caveolae, which have diverse functions. Our recent study on the temporomandibular joint (TMJ) revealed expressions of Cav1 and muscle-specific Cav3 in some synovial fibroblast-like type B cells with well-developed caveolae. However, the involvement of Cav3 expression in the differentiation and maturation of type B cells remains unclear. The present study therefore examined the chronological alterations in the localization of Cav3 in the synovial lining cells of the rat TMJ during postnatal development by immunocytochemical techniques. Observations showed immature type B cells possessed a few caveolae with Cav1 but lacked Cav3 protein at postnatal day 5 (P5). At P14, Cav3-immunopositive type B cells first appeared in the synovial lining layer. They increased in number and immunointensity from P14 to P21 as occlusion became active. In immunoelectron microscopy and double immunolabeling with heat shock protein 25 (Hsp25) and Cav3, coexpressed type B cells developed rough endoplasmic reticulum and numerous caveolae, while the Cav3-immunonegative type B cell with Hsp25 immunoreaction possessed few of these. Results suggest that Cav3 expression, which is closely related to added functional stimuli, reflects the differentiation of the type B synoviocytes.

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  • Histological and elemental analyses of impaired bone mineralization in klotho-deficient mice 査読

    Hironobu Suzuki, Norio Amizuka, Kimimitsu Oda, Masaki Noda, Hayato Ohshima, Takeyasu Maeda

    JOURNAL OF ANATOMY   212 ( 3 )   275 - 285   2008年3月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:BLACKWELL PUBLISHING  

    The klotho gene-deficient mouse is known as an animal model for an accelerated gerontic state, mimicking osteoporosis, skin atrophy, ectopic calcification, and gonadal dysplasia. To elucidate the influence of klotho deficiency on bone mineralization, we examined the ultrastructures of osteoblasts and bone matrices in addition to performing the elemental mapping of calcium, phosphorus, and magnesium in the bone. Under anesthesia, 4- and 5-week-old klotho-deficient mice (klotho(-/-) mice) and their wild-type littermates were perfused with either 4% paraformaldehyde for light microscopic observation or 4% paraformaldehyde and 0.0125% glutaraldehyde for electron microscopic observation. The femurs and tibiae were processed for both observations. Paraffin sections were subject to alkaline phosphatase and tartrate resistant acid phosphatase histochemistry. Semithin and ultrathin sections obtained from epoxy resin-embedded specimens were used for detecting mineralization - according to von Kossa's staining method - and for elemental mapping by electron probe micro-analyzer, respectively. Alkaline phosphatase-positive plump osteoblasts adjacent to the growth plate normally developed cell organelles in the klotho(-/-) metaphyses. This, however, contrasted with the flattened osteoblasts covering the metaphyseal trabeculae and accompanied by small tartrate resistant acid phosphatase-positive osteoclasts. The wild-type mice displayed the mineralized matrix at the zone of hypertrophic chondrocyte of the growth plate and well-mineralized metaphyseal trabeculae parallel to the longitudinal axis of the bone. Alternatively, the klotho(-/-) mice demonstrated a thick mineralized matrix from the proliferative zone of the growth plate as well as the large non-mineralized area in the metaphyseal trabeculae. Consistently, electron probe micro-analysis verified sporadic distributions of higher or lower concentrations of calcium and phosphorus in each trabecule of the klotho(-/-) mice. The distribution of magnesium, however, was almost uniform. Under transmission electron microscopy, osteoblasts on the metaphyseal trabeculae displayed less-developed cell organelles in the klotho(-/-) mice. Thus, the klotho deficiency appears not only to reduce osteoblastic population, but also to disturb bone mineralization.

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  • Functional requirement of CCN2 for intramembranous bone formation in embryonic mice 査読

    Harumi Kawaki, Satoshi Kubota, Akiko Suzuki, Tomohiro Yamada, Tatsushi Matsumura, Toshiko Mandal, Mayumi Yao, Takeyasu Maeda, Karen M. Lyons, Masaharu Takigawa

    BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS   366 ( 2 )   450 - 456   2008年2月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:ACADEMIC PRESS INC ELSEVIER SCIENCE  

    CCN2 is best known as a promoter of chondrocyte differentiation among the CCN family members, and Ccn2 null mutant mice display skeletal dysmorphisms. However, little is known concerning the roles of CCN2 during bone formation. We herein present a comparative analysis of wild-type and Ccn2 null mice to investigate the roles of CCN2 in bone development. Multiple histochemical methods were employed to analyze the effects of CCN2 deletion in vivo, and effects of CCN2 on the osteogenic response were evaluated with the isolated and cultured osteoblasts. As a result, we found a drastic reduction of the osteoblastic phenotype in Ccn2 null mutants. Importantly, addition of exogenous CCN2 promoted every step of osteoblast differentiation and rescued the attenuated activities of the Ccn2 null osteoblasts. These results suggest that CCN2 is required not only for the regulation of cartilage and subsequent events, but also for the normal intramembranous bone development. (c) 2007 Elsevier Inc. All rights reserved.

    DOI: 10.1016/j.bbrc.2007.11.155

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  • Histological evaluation on bone regeneration of dental implant placement sites grafted with a self-setting alpha-tricalcium phosphate cement 査読

    Masayoshi Nakadate, Norio Amizuka, Minqi Li, Paulo H. L. Freitas, Kimimitsu Oda, Shuichi Nomura, Katsumi Uoshima, Takeyasu Maeda

    MICROSCOPY RESEARCH AND TECHNIQUE   71 ( 2 )   93 - 104   2008年2月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:WILEY-LISS  

    This study aimed to evaluate the histological characteristics of the new bone formed at dental implant placement sites concomitantly grafted with a self-setting tricalcium phosphate cement (BIOPEX-R (R)). Standardized defects were created adjacent to the implants in maxillae of 4-week-old male Wistar rats, and were concomitantly filled with BIOPEX-R (R). Osteogenesis was examined in two sites of extreme clinical relevance: (1) the BIOPEX-R (R)-grafted surface corresponding to the previous alveolar ridge (alveolar ridge area), and (2) the interface between the grafting material and implants (interface area). At the alveolar ridge area, many tartrate-resistant acid phosphatase (TRAPase)-reactive osteoclasts had accumulated on the BIOPEX-R (R) surface and were shown to migrate toward the implant. After that, alkaline phosphatase (ALPase)-positive osteoblasts deposited new bone matrix, demonstrating their coupling with osteoclasts. On the other hand, the interface area showed several osteoclasts initially invading the narrow gap between the implant and graft material. Again, ALPase-positive osteoblasts were shown to couple with osteoclasts, having deposited new bone matrix after bone resorption. Transmission electron microscopic observations revealed direct contact between the implant and the new bone at the interface area, although few thin cells could still be identified. At both the alveolar ridge and the interface areas, newly formed bone resembled compact bone histologically. Also, concentrations of Ca, P, and Mg were much alike with those of the preexistent cortical bone. In summary, when dental implant placement and grafting with BIOPEX-R (R) are done concomitantly, the result is a new bone that resembles compact bone, an ideal achievement in reconstructive procedures for dental implantology.

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  • Involvement of the klotho protein in dentin formation and mineralization 査読

    Hironobu Suzuki, Norio Amizuka, Kimimitsu Oda, Masaki Noda, Hayato Ohshima, Takeyasu Maeda

    ANATOMICAL RECORD-ADVANCES IN INTEGRATIVE ANATOMY AND EVOLUTIONARY BIOLOGY   291 ( 2 )   183 - 190   2008年2月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:WILEY-LISS  

    Klotho-deficient mice exhibit multiple pathological conditions resembling human aging. Our previous study showed alterations in the distribution of osteocytes and in the bone matrix synthesis in klotho-deficient mice. Although the bone and tooth share morphological features such as mineralization processes and components of the extracellular matrix, little information is available on how klotho deletion influences tooth formation. The present study aimed to elucidate the altered histology of incisors of klotho-deficient mice-comparing the findings with those from their wild-type litter-mates, by using immunohistochemistry for alkaline phosphatase (ALP), osteopontin, and dentin matrix protein-1 (DMP-1), terminal deoxynucleotidyl transferase-mediated deoxyuridinetriphosphate nick end-labeling (TUNEL) detection for apoptosis, and electron probe microanalyzer (EPMA) analysis on calcium (Ca), phosphate (P), and magnesium (Mg). Klotho-deficient incisors exhibited disturbed layers of odontoblasts, predentin, and dentin, resulting in an obscure dentin-predentinal border at the labial region. Several odontoblast-like cells without ALP activity were embedded in the labial dentin matrix, and immunopositivity for DMP-1 and osteopontin was discernible in the matrix surrounding these embedded odontoblast-like cells. TUNEL detection demonstrated an apoptotic reaction in the embedded odontoblast-like cells and pulpal cells in the klotho-deficient mice. EPMA revealed lower concentrations of Ca, P, and Mg in the klotho-deficient dentin, except for the dentin around abnormal odontoblast-like cells. These findings suggest the involvement of the klotho gene in dentinogenesis and its mineralization.

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  • The effect of neonatal capsaicin treatment on the CGRP-immunoreaction in the trigeminal subnucleus caudalis of mice 査読

    Megumi Aita, Takeyasu Maeda, Kenji Se

    BIOMEDICAL RESEARCH-TOKYO   29 ( 1 )   33 - 42   2008年2月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:BIOMEDICAL RESEARCH PRESS LTD  

    The calcitonin-gene related peptide (CGRP) is a primary afferent neurotransmitter in the trigeminal system. Although a neonatal administration of capsaicin eliminates substance P (SP)-mediated nociceptive responses to induce a permanent functional reduction in C-fibers, little information is available regarding changes in CGRP-immunoreaction in mice undergoing neonatal capsaicin treatment (CP mice). This study examined postnatal changes in the distribution of CGRP-immunoreaction in the trigeminal subnucleus caudalis and trigeminal ganglion of CP mice by immunohistochemical technique and a quantitative analysis. Immunohistochemistry for CGRP in the subnucleus caudalis (Vc) demonstrated two dense distributions of neurons in the CP mice as well as naive mice: in the marginal layer and the region 400-600 mu m deep. The quantitative analysis revealed no significant difference in the density of CGRP immunoreaction between naive and CP mice 1-8 weeks of age. In the trigeminal ganglion of both groups, the size distribution of CGRP-positive neurons displayed a distribution pattern with one peak in 200-300 mu m(2) at week 1 and with two peaks in 200-300 mu m(2) and 600-700 mu m(2) at week 8 but no significant difference in neural density existed between these regions. When double staining in the naive mice with CGRP or SP and VR1, a capsaicin receptor, was done, many trigeminal ganglion neurons co-expressed SP- and VR1-immunoreactions, but rarely exhibited CGRP/VR1-co-localization. Taken together with previous data, these current observations suggest that CGRP containing afferent neurons possibly performs differing roles in nociceptive afferent input transmission within the Vc from SP-containing neurons in mice.

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  • 生涯学習力に対するPBLチュートリアルの効果 査読

    小野和宏, 八木 稔, 大内章嗣, 魚島勝美, 安島久雄, 林 孝文, 齋藤 功, 興地隆史, 前田健康

    日歯教誌   24 ( 1月2日 )   145 - 149   2008年

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  • Histological examination of bone regeneration achieved by combining grafting with hydroxyapatite and thermoplastic bioresorbable plates 査読

    Taku Kojima, Norio Amizuka, Akiko Suzuki, Paulo Henrique, Luiz de Freitas, Michiko Yoshizawa, Akira Kudo, Chikara Saito, Takeyasu Maeda

    JOURNAL OF BONE AND MINERAL METABOLISM   25 ( 6 )   361 - 373   2007年11月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:SPRINGER TOKYO  

    In this study, we present a novel guided bone regeneration (GBR) concept that consists of combining Boneject, a bone substitute containing atelocollagen and bovine hydroxyapatite particles, with thermoplastic, bioresorbable plates (DeltaSystem) known to resist mechanical loading. In rat calvariae, standardized bone defects were filled with Boneject and covered with a convex DeltaSystem plate. Tissue from rats at 1, 2, 4, 8, and 12 weeks postoperation were fixed with an aldehyde solution, and the new bone formed at the defects was histologically assessed. At 1 week, alkaline phosphatase (ALP)-negative cells deriving from the bottom region of the defect could be found up to half the height of the cavity. Boneject particle surfaces in the bottom region revealed an intense osteopontin immunopositivity whereas those in the upper region did not. Tartrate-resistant acid phosphatase (TRAP)-positive osteoclasts accumulated on the surfaces of osteopontin-coated particles. A newly formed, woven-like bone featuring ALP-positive osteoblasts extended from the native bone. At the second week, the newly formed woven bone had surrounded the Boneject particles. Cement lines, which indicate active bone remodeling, could be observed in the new bone despite its immaturity. Four weeks after surgery, the new bone had reached the height of the DeltaSystem plate, and just beneath it a periostin-positive fibrous layer covered the mix of new bone and Boneject particles. By then, despite having acceptable histological features, electron probe microanalyzer (EPMA) and transmission electron microscope (TEM) analyses revealed that the new bone could not be regarded as compact bone. At 8 and 12 weeks, the new bone showed compact bone-like features according to TEM and EPMA assessments. Summarizing, the combination of a bone substitute such as Boneject and a rigid, bioresorbable plate appears to be osteoconductive and to promote bone remodeling, leading to the genesis of a tissue similar to the one that is regarded as the "gold standard" for bone regeneration: the compact bone.

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  • The expression and production of vascular endothelial growth factor in oral mucosa equivalents 査読

    Y. Nakanishi, K. Izumi, M. Yoshizawa, C. Saito, Y. Kawano, T. Maeda

    INTERNATIONAL JOURNAL OF ORAL AND MAXILLOFACIAL SURGERY   36 ( 10 )   928 - 933   2007年10月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:CHURCHILL LIVINGSTONE  

    Angiogenesis is anticipated during wound healing. Vascular endothelial growth factor (VEGF) is a potent direct angiogenic factor that stimulates endothelial cell migration and proliferation in vitro and angiogenesis in vivo. Ex vivo produced oral mucosa equivalent (EVPOME) grafts have been reported to promote revascularization in the underlying tissue after grafting. The aim of this study was to evaluating the following: VEGF mRNA and its protein expression in EVPOME grafts, the protein levels in conditioned media produced by EVPOMEs, and the ability of VEGF to stimulate the growth of microvascular endothelial cells. VEGF mRNA expression and immunoreaction were found in basal and suprabasal layers. VEGF secreted by EVPOME was detected throughout the period of manufacture of the grafts, and became elevated for the first week at an air-liquid interface. Both EVPOME-conditioned media and a medium containing 10 ng/mL recombinant human VEGF induced endothelial cell proliferation, while neutralization of VEGF by an antibody blocked cell growth. These results suggest that VEGF secreted by EVPOME grafts might assist initial vascularization after grafting, which is critical to subsequent graft survival.

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  • Histological assessments on the abnormalities of mouse epiphyseal chondrocytes with short term centrifugal loading 査読

    Paulo Henrique Luiz de Freitas, Taku Kojima, Soblian Ubaidus, Minqi Li, Guangwei Shang, Ritsuo Takagi, Takeyasu Maeda, Kimimitsu Oda, Hidehiro Ozawa, Norio Amizuka

    BIOMEDICAL RESEARCH-TOKYO   28 ( 4 )   191 - 203   2007年8月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:BIOMEDICAL RESEARCH PRESS LTD  

    We have examined the morphological changes in chondrocytes after exposure to experimental hypergravity. Tibial epiphyseal cartilages of 17-days-old mouse fetuses were exposed to centrifugation at 3G for 16 It mimicking hypergravitational environment (experimental group), or subjected to stationary cultures (control group). Centrifugation did not affect the sizes of epiphyseal cartilage, chondrocyte proliferation, type X collagen-positive hypertrophic zone, and the mRNA expressions of parathyroid hormone-related peptide and fibroblast growth factor receptor Ill. However, centrifuged chondrocytes showed abnormal morphology and aberrant spatial arrangements, resulting in disrupted chondrocytic columns. Through histochemical assessments, actin filaments were shown to distribute evenly along cell membranes of control proliferative chondrocytes, while chondrocytes subjected to centrifugal force developed a thicker layer of actin filaments. Transmission electron microscopic observations revealed spotty electron-dense materials underlying control chondrocytes' cell membranes, while experimental chondrocytes showed their thick layer. In the intracolumnar regions of the control cartilage, longitudinal electron-dense fibrils were associated with short cytoplasmic processes of normal chondrocytes, indicating assumed cell-to-matrix interactions. These extracellular fibrils were disrupted in the centrifuged samples. Summarizing, altered actin filaments associated with cell membranes, irregular cell shape and disappearance of intracolumnar extracellular fibrils suggest that hypergravity disturbs cell-to-matrix interactions in our cartilage model.

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  • Histochemical examinations on cortical bone regeneration induced by thermoplastic bioresorbable plates applied to bone defects of rat calvariae 査読

    Taku Kojima, Paulo H. L. Freitas, Sobhan Ubaidus, Akiko Suzuki, Minqi Li, Michiko Yoshizawai, Kimimitsu Oda, Takeyasu Maeda, Akira Kudo, Chikara Saito, Norio Amizuka

    BIOMEDICAL RESEARCH-TOKYO   28 ( 4 )   219 - 229   2007年8月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:BIOMEDICAL RESEARCH PRESS LTD  

    We aimed to histologically elucidate whether bioresorbable plates (DeltaSystem (R)) can induce cortical bone formation, which is essential for long-lasting bone augmentation. Standardized bone defects in rat calvariae were covered with a convexly-shaped DeltaSystem (R) plate, and then processed for histological observations. At 1 week, alkaline phosphatase-positive osteoblasts were seen in the newly-formed bone extending from the cavity's bottom, indicating accelerated osteogenesis. A thick layer of soft connective tissue positive for periostin, a hallmark of periosteum, covered this new bone. At 2 weeks, a spongy bone had filled the cavity up to half its height. The inner layer of the soft tissue facing the spongy bone revealed abundant periostin and osteopontin, and had many tartrate-resistant acid phosphatase-positive osteoclasts. At 4 weeks, this layer had given rise to thin new bony matrices without relation to the spongy bone arising from the cavity. These bone matrices had been thickened by 8 weeks, and turned into a thick cortical bone outlining the regenerated bone at 12 weeks. Thus, our study has provided histological evidences of cortical osteogenesis when DeltaSystem (R) plates are used for bone augmentation procedures.

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  • 老化モデルマウス顎下腺への男性ホルモンの効果

    鈴木 啓展, 網塚 憲生, 天野 修, 前田 健康

    Journal of Oral Biosciences   49 ( Suppl. )   110 - 110   2007年8月

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    記述言語:日本語   出版者・発行元:(一社)歯科基礎医学会  

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  • Immunolocalization of aquaporin-1 in the mechanoreceptive Ruffini endings in the periodontal ligament 査読

    Batheegama Gammacharige Tharanga Lakmali Nandasena, Akiko Suzuki, Megumi Aita, Yoshiro Kawano, Kayoko Nozawa-Inoue, Takeyasu Maeda

    BRAIN RESEARCH   1157   32 - 40   2007年7月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:ELSEVIER SCIENCE BV  

    Previous ultrastructural studies have suggested an axon-Schwarm cell interaction in the periodontal Ruffini ending, a primary mechanoreceptor. However, no information is available on the transport mechanism between them. The present study examined the immunolocalization of aquaporin-1 (AQP1) and -4 (AQP4), a member of the water-selective channel, in the periodontal Ruffini endings of the rat incisors and trigeminal ganglion. In addition, the expression of mRNA for AQP1 and 4 was detected in the trigeminal ganglion by a RT-PCR technique. A single PCR product of the sizes anticipated for AQP1 and 4 was detectable in a reverse transcripted cDNA sample from the trigeminal ganglion, whose neurons innervate the periodontal Ruffini endings. An AQP1 immunoreaction was recognizable in the axon terminals of the periodontal Ruffini endings as well as their associated terminal Schwann cells, as confirmed with a double staining with AQP1 and either PGP9.5 or S-100 protein. However, no immunoreaction for AQP4 was found in periodontal Ruffini endings. Although the AQP4 immunoreaction was localized in some satellite cells - but never in neurons - of the trigeminal ganglion, 16.1% trigeminal neurons showed the AQP1 immunoreaction. Furthermore, the AQP1 immunoreaction was found in certain satellite cells which surrounded AQP1-positive or -negative neurons. An analysis of a cross-sectional area of these positive neurons demonstrated that approximately 66.9% of the positive neurons were 400-1000 mu m(2) (671.4 +/- 172.4 mu m(2)), indicating that they could be categorized as medium-sized neurons which mediate mechanotransduction. These findings suggest that AQP1 controls water transport in the periodontal Ruffini endings. (c) 2007 Elsevier B.V. All rights reserved.

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  • 骨細胞の分布/陰窩-細管系に関する組織学的評価(Histochemical assessments on the distribution of osteocytes / lacunar canalicular system)

    Ubaidus Sobhan, 李 敏啓, 小島 拓, 織田 公光, 前田 健康, 高木 律男, 網塚 憲生

    日本骨代謝学会学術集会プログラム抄録集   25回   203 - 203   2007年6月

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    記述言語:英語   出版者・発行元:(一社)日本骨代謝学会  

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  • Involvement of neurotrophin-4/5 in regeneration of the periodontal Ruffini endings at the early stage 査読

    Shahiqul Jabbar, Fumiko Harada, Megumi Aita, Megumi Ohishi, Isao Saito, Yoshiro Kawano, Akiko Suzuki, Kayoko Nozawa-Inoue, Takeyasu Maeda

    JOURNAL OF COMPARATIVE NEUROLOGY   501 ( 3 )   400 - 412   2007年3月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:WILEY-LISS  

    Little is known about the role of neurotrophin-4/5 (NT-4/5) in the regeneration of mechano-receptors. Therefore, the present study examined the regeneration process of Ruffini endings in the periodontal ligament in nt-4/5-deficient and wildtype mice following transection of the inferior alveolar nerve by immunohistochemistry for protein gene product 9.5 (PGP 9.5), a general neuronal marker, and by computer-assisted quantitative image analysis. Furthermore, rescue experiments by a continuous administration of recombinant NT-4/5 were performed and analyzed quantitatively. At postoperative day 3 (PO 3d), almost all PGP 9.5-positive neural elements had disappeared; they began to appear in both types of animals at PO 7d. At PO 10d, almost all nerve fibers showed a beaded appearance, with fewer ramifications in both types of mice. Although the regeneration proceeded in the wildtype, a major population of the periodontal Ruffini endings continued to display smooth outlines at PO 28d in the nt-4/5 homozygous mice. The reduction ratio of neural density reached a maximum at PO 3d, decreased at PO 10d, and later showed a plateau. In a rescue experiment, an administration of NT-4/5 showed an acceleration of nerve regeneration in the homozygous mice. These findings indicate that the nt-4/5depletion causes a delay in the regeneration of the periodontal Ruffini endings, but the delay is shortened by an exogenous administration of NT-4/5. Combined with our previous findings of bdnf-deficient mice (Harada et al. [2003] Arch Histol Cytol 66:183-194), these morphological and numerical data suggest that multiple neurotrophins such as NT-4/5 and brain-derived neurotrophic factor (BDNF) play roles in their regeneration in a stage-specific manner.

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  • Expression of caveolin-3 in the fibroblast-like type B synoviocytes in the rat temporomandibular joint 査読

    Kayoko Nozawa-Inoue, Akiko Suzuki, Masahiro Niwano, Yoshiro Kawano, Takeyasu Maeda

    ANATOMICAL RECORD-ADVANCES IN INTEGRATIVE ANATOMY AND EVOLUTIONARY BIOLOGY   290 ( 3 )   238 - 242   2007年3月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:WILEY-LISS  

    The present study revealed that the fibroblast-like type B synoviocytes (covering the surface of the synovial membrane in the rat temporomandibular joint) had muscle-specific caveolin-3 protein in their caveolae. The existence of two kinds of type B synoviocytes (with and without caveolin-3-immunoreactions even in the synovial lining layer) might reflect the functional difference between them.

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  • Expression of caveolin-3 in the fibroblast-like type B synoviocytes in the rat temporomandibular joint 査読

    Kayoko Nozawa-Inoue, Akiko Suzuki, Masahiro Niwano, Yoshiro Kawano, Takeyasu Maeda

    Anatomical Record   290 ( 3 )   238 - 242   2007年3月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    The present study revealed that the fibroblast-like type B synoviocytes (covering the surface of the synovial membrane in the rat temporomandibular joint) had muscle-specific caveolin-3 protein in their caveolae. The existence of two kinds of type B synoviocytes (with and without caveolin-3-immunoreactions even in the synovial lining layer) might reflect the functional difference between them. © 2007 Wiley-Liss, Inc.

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  • Involvement of GDNF and its receptors in the maturation of the periodontal Ruffini endings 査読

    Yasushi Igarashi, Megumi Aita, Akiko Suzuki, Tharanga Nandasena, Yoshiro Kawano, Kayoko Nozawa-Inoue, Takeyasu Maeda

    NEUROSCIENCE LETTERS   412 ( 3 )   222 - 226   2007年2月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:ELSEVIER IRELAND LTD  

    Our recent study revealed an intense immunoreaction for GDNF and its receptors in the Ruffini endings, primary mechanoreceptors in the periodontal ligament, of young rats. However, no information is available for the expression of GDNF and its receptors during their development. The present study aimed to reveal postnatal changes in the immuno-expression of GDNF, GFR alpha l and RET in the periodontal Ruffini endings of the rat incisors by double immunofluorescent staining. At postnatal day 3 (PO 3d), no structure with GDNF-, GFR alpha l-, or RET-immunoreaction existed in the periodontal ligament. The PGP 9.5-positive nerve fibers without GDNF- and RET-immunoreaction displayed a dendritic fashion at PO lw, with a GFR alpha l-reaction found around these nerves. At PO 2w, GDNF-positive terminal Schwann cells occurred near the thick and dendritic axons, a part of which showed a RET-reaction, with no reactive cells near the thin nerves. The terminal Schwann cells became positive for GFR alpha l, but lacked RET-immunoreaction. At PO 3w, when the formation of the periodontal Ruffini endings had proceeded, GDNF-positive terminal Schwann cells began to increase in number. This stage-specific immuno-expression pattern suggests that GDNF is a key molecule for the maturation and maintenance of the periodontal Ruffini endings. (c) 2006 Elsevier Ireland Ltd. All rights reserved.

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  • An evaluation of problem-based learning course at Niigata University Faculty of Dentisitry 招待 査読

    Maeda T, Ono K, Ohuchi A, Hayashi T, Saito I, Okiji T, Uoshima K

    Dentistry in Japan   43   166 - 171   2007年

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  • 軟骨内骨化のMMP-9の役割における組織学的検討

    小島 拓, 李 敏啓, 齊藤 力, 前田 健康, 網塚 憲生

    Journal of Oral Biosciences   48 ( Suppl. )   126 - 126   2006年9月

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    記述言語:日本語   出版者・発行元:(一社)歯科基礎医学会  

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  • MMP-9遺伝子欠損マウスにおける軟骨内骨化異常の組織化学的検討

    小島 拓, 網塚 憲生, 李 敏啓, 小崎 直人, 齊藤 力, 高石 官成, 前田 健康

    日本骨代謝学会学術集会プログラム抄録集   24回   219 - 219   2006年7月

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    記述言語:日本語   出版者・発行元:(一社)日本骨代謝学会  

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  • Localization of CD44 and hyaluronan in the synovial membrane of the rat temporomandibular joint 査読

    A Suzuki, K Nozawa-Inoue, N Amizuka, K Ono, T Maeda

    ANATOMICAL RECORD PART A-DISCOVERIES IN MOLECULAR CELLULAR AND EVOLUTIONARY BIOLOGY   288A ( 6 )   646 - 652   2006年6月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:WILEY-LISS  

    Previous studies have pointed out a lack of adhesion structures in the synovial lining layer of the rat temporomandibular joint (TMJ) despite showing an epithelial arrangement. CD44, a major cell adhesion molecule, plays crucial roles as an anchor between cells and extracellular matrices by binding hyaluronan (HA) for the development of organs or the metastasis of tumors. The present study examined the localization of CD44 in the synovial membrane of the rat TMJ by immunocytochemistry for OX50, ED1, and Hsp25, which are markers for the rat CD44, macrophage-like type A, and fibroblast-like type B synoviocytes, respectively. Histochemistry for HA-binding protein (HABP) was also employed for the detection of HA. OX50 immunoreactions were found along the cell surface and, in particular, accumulated along the surface of the articular cavity. Observations by a double immunostaining and immunoelectron microscopy revealed that all the OX50-immunopositive cells were categorized as fibroblastic type B cells, which had many caveolae and a few vesicles reactive to intense OX50. However, the macrophage-like type A cells did not have any OX50 immunoreaction in the synovial lining layer. A strong HABP reaction was discernable in the extracellular matrix surrounding both OX50-positive and -negative cells in the synovial lining layers, exhibiting a meshwork distribution, but weak in its sublining layer. This localization pattern of CD44 and HABP might be involved in the formation of the epithelial arrangement of the synovial lining layer. Furthermore, OX50 immunonegativity in the type A cells suggests their low phagocytotic activity in the rat TMJ under normal conditions.

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  • Histological and immunohistochemical changes in the submandibular gland in klotho-deficient mice 査読

    Hironobu Suzuki, Norio Amizuka, Masaki Noda, Osamu Aman, Takeyasu Maeda

    ARCHIVES OF HISTOLOGY AND CYTOLOGY   69 ( 2 )   119 - 128   2006年6月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:INT SOC HISTOLOGY & CYTOLOGY  

    The submandibular gland (SMG) has been regarded as an age-stable organ in spite of reports on its structural changes with aging. Although the klotho gene is involved in aging, little information is available regarding its effects on morphological changes of SMGs. The present study examined the histological and immunohistochemical features of SMGs in klotho-deficient mice - which are well-established aging models - by immunohistochemical and histochemical techniques. Five kinds of cellular markers - against NGF, EGF, Mn- and Cu/Zn-SOD, and RITC-conjugated phalloidin were used for the identification of cell types. In klotho-deficient mice, the SMGs lost their granular ducts and each lobe diminished. The granular duct showed strong immunoreactivities for NGF and EGF in the wild-type mice, but the NGF- and EGF-immunopositive ducts decreased in number remarkably in klotho-deficient mice. Interestingly, instead of a loss of the granular duct, the striated duct located on the distal portion in the homozygous mice came to show NGF- and EGF-immunoreactions. Neither Mn- and Cu/Zn-SOD immunoreactivities in the duct system nor the phalloidin-reaction in the myoepithelial cells differed between the wild-type and klotho-deficient mice. Our findings suggest that the klotho gene inhibited the differentiation of the granular duct from the striated duct due to the repression and/or down-regulation of sexual and growth hormones.

    DOI: 10.1679/aohc.69.119

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  • Expression of GDNF and its receptors in the periodontal mechanoreceptor 査読

    Megumi Aita, Yoshiro Kawano, Takeyasu Maeda

    NEUROSCIENCE LETTERS   400 ( 1-2 )   25 - 29   2006年5月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:ELSEVIER IRELAND LTD  

    Our previous studies have revealed the involvement of signaling pathways of BDNF and NT-4/5 via TrkB in the development, regeneration, survival and maintenance of the Ruffini endings, primary mechanoreceptors in the periodontal ligament. However, the involvement of other neurotrophins remains unclear. The present study examined the expression of GDNF, GFR alpha 1, and RET in the incisor periodontal ligament and trigeminal ganglion of young rats by RT-PCR and immunocytochemistry. All these mRNAs were detected in both tissues by RT-PCR. These immunoreactions were found in the terminal Schwann cells associated with the periodontal Ruffini endings, as confirmed by histochemistry for non-specific cholinesterase activity. Their axonal branches showed GF alpha 1- and RET-immunoreactions but lacked GDNF-immunoreactivity. In the trigeminal ganglion. about 30% of the neurons were immunoreactive to GFR alpha 1 and RET. Averages of cross-sectional areas of their positive neurons demonstrated that they could mainly be categorized as medium-sized neurons. GDNF-immunoreaction was restricted to the satellite cells and not in trigeminal ganglion neurons. These findings indicate that GDNF mediates trophic effects on the survival and target innervation of the periodontal Ruffini endings via GFR alpha 1 and RET. (c) 2006 Elsevier Ireland Ltd. All rights reserved.

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  • 生後マウスにおける下顎頭軟骨の経時的観察

    池田 順行, Hossain Kazi Sazzad, 井上 佳世子, 鈴木 晶子, 高木 律男, 前田 健康, 網塚 憲生

    日本顎関節学会雑誌   18 ( 1 )   42 - 42   2006年4月

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  • Histochemical evidence of osteoclastic degradation of extracellular matrix in osteolytic metastasis originating from human lung small carcinoma (SBC-5) cells 査読

    MQ Li, N Amizuka, K Takeuchi, PHL Freitas, Y Kawano, M Hoshino, K Oda, K Nozawa-Inoue, T Maeda

    MICROSCOPY RESEARCH AND TECHNIQUE   69 ( 2 )   73 - 83   2006年2月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:WILEY-LISS  

    The aim of this study was to assess the dynamics of osteoclast migration and the degradation of unmineralized extracellular matrix in an osteolytic metastasis by examining a well-standardized lung cancer metastasis model of nude mice. SBC-5 human lung small carcinoma cells were injected into the left cardiac ventricle of 6-week-old BALB/c nu/nu mice under anesthesia. At 25-30 days after injection, the animals were sacrificed and their femora and/or tibiae were removed for histochemical analyses. Metastatic lesions were shown to occupy a considerable area extending from the metaphyses to the bone marrow region. Tartrate resistant acid phosphatase (TRAPase)-positive osteoclasts were found in association with an alkaline phosphatase (ALPase)positive osteoblastic layer lining the bone surface, but could also be localized in the ALPase-negative stromal tissues that border the tumor nodules. These stromal tissues were markedly positive for osteopontin, and contained a significant number of TRAPase-positive osteoclasts expressing immunoreactivity for CD44. We thus speculated that, mediating its affinity for CD44, osteopontin may serve to facilitate osteoclastic migration after their formation associated with ALPase-positive osteoblasts. We next examined the localization of cathepsin K and matrix metallo-proteinase-9 (MMP-9) in osteoclasts. Osteoclasts adjacent to the bone surfaces were positive for both proteins, whereas those in the stromal tissues in the tumor nests showed only MMP-9 immunoreactivity. Immunoelectron microscopy disclosed the presence of MMP-9 in the Golgi apparatus and in vesicular structures at the baso-lateral cytoplasmic region of the osteoclasts found in the stromal tissue. MMP-9-positive vesicular structures also contained fragmented extracellular materials. Thus, osteoclasts appear to either select an optimized function, namely secreting proteolytic enzymes from ruffled borders during bone resorption, or recognize the surrounding extracellular matrix by mediating osteopontin/CD44 interaction, and internalize the extracellular matrices.

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  • Expression of caveolin-1 in the rat temporomandibular joint 査読

    Nozawa-Inoue Kayoko, Akiko Suzuki, Norio Amizuka, Takeyasu Maeda

    Anatomical Record - Part A Discoveries in Molecular, Cellular, and Evolutionary Biology   288 ( 1 )   8 - 12   2006年1月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    This immunocytochemical study revealed the expression of caveolin-1, a major protein of caveolae, in the rat temporomandibular joint. In the synovial lining layer, immunoreactive products for caveolin-1 were detected on the cell membrane of the fibroblast-like type B cells, as confirmed by immunocytochemistry for heat shock protein 25. The cells in the articular disk, the articular layer, and zone of proliferation of the mandibular condyle also showed intense immunoreactions for caveolin-1. © 2005 Wiley-Liss, Inc.

    DOI: 10.1002/ar.a.20275

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  • Expression of caveolin-1 in the rat temporomandibular joint 査読

    K Nozawa-Inoue, A Suzuki, N Amizuka, T Maeda

    ANATOMICAL RECORD PART A-DISCOVERIES IN MOLECULAR CELLULAR AND EVOLUTIONARY BIOLOGY   288A ( 1 )   8 - 12   2006年1月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:WILEY-LISS  

    This immunocytochemical study revealed the expression of caveolin-1, a major protein of caveolae, in the rat temporomandibular joint. In the synovial lining layer, immunoreactive products for caveolin-1 were detected on the cell membrane of the fibroblast-like type B cells, as confirmed by immunocytochemistry for heat shock protein 25. The cells in the articular disk, the articular layer, and zone of proliferation of the mandibular condyle also showed intense immunoreactions for caveolin-1. (C) 2005 Wiley-Liss, Inc.

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  • 顎関節関節腔形成における血管内皮細胞とマクロファージの動態

    鈴木 晶子, 野澤 佳世子[井上], 網塚 憲生, 前田 健康

    新潟歯学会雑誌   35 ( 2 )   246 - 246   2006年1月

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  • チタンインプラント植立時に併用したα-TCP系自己硬化型補填材が骨再生に及ぼす影響

    中舘 正芳, 網塚 憲生, 野村 修一, 前田 健康

    新潟歯学会雑誌   35 ( 2 )   265 - 265   2006年1月

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  • 骨芽細胞の活性に及ぼす破骨細胞の影響(第二報) c-src遺伝子欠損マウスを用いた組織化学的検討

    坂上 直子, 網塚 憲生, 李 敏啓, Freitas Paulo H.L., 前田 健康

    新潟歯学会雑誌   35 ( 2 )   257 - 257   2006年1月

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  • ポリ乳酸プレートおよびhydroxyapatite補填材併用による骨再生の組織化学的検索

    小島 拓, 網塚 憲生, 鈴木 晶子, 芳澤 享子, 齊藤 力, 前田 健康

    新潟歯学会雑誌   35 ( 2 )   264 - 264   2006年1月

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  • 骨芽細胞の活性に及ぼす破骨細胞の影響(第一報) op/opマウスを用いた組織化学的検討

    坂上 直子, 網塚 憲生, 李 敏啓, 前田 健康

    新潟歯学会雑誌   35 ( 2 )   257 - 257   2006年1月

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  • 歯科医学教育へのPBLテュートリアルの導入 査読

    小野和宏, 大内章嗣, 魚島勝美, 林 孝文, 西山秀昌, 安島久雄, 小林正治, 瀬尾憲司, 齋藤 功, 程 ?, 山田好秋, 前田健康

    日歯教誌   22 ( 1 )   58 - 71   2006年

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  • Neurotrophin-4/5-depletion induces a delay in maturation of the periodontal Ruffini endings in mice 査読

    Y Maruyama, F Harada, S Jabbar, Saito, I, M Aita, Y Kawano, A Suzuki, K Nozawa-Inoue, T Maeda

    ARCHIVES OF HISTOLOGY AND CYTOLOGY   68 ( 4 )   267 - 288   2005年12月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:INT SOC HISTOLOGY & CYTOLOGY  

    Neurotrophin-4/5 (NT-4/5)-a member of the neurotrophic factors-is a ligand for TrkB, which has been reported to be expressed in the mechanoreceptive Ruffini endings of the periodontal ligament. The present study examined developmental changes in the terminal morphology and neural density in homozygous mice with a targeted disruption of the nt-4/5 gene and wild-type mice by immunohistochemistry for protein gene product 9.5 (PGP 9.5), a general neuronal marker, and by quantitative analysis using an image analyzer. Postnatal development of terminal Schwann cells was also investigated by enzymatic histochemistry for non-specific cholinesterase activity (ChE). Furthermore, the immuno-expression of TrkB and low affinity nerve growth factor receptor (p75-NGFR) was surveyed in the periodontal Ruffini endings as well as trigeminal ganglion. At postnatal 1 week, the lingual periodontal ligament of both types of mice contained PGP 9.5-positive nerve fibers showing a tree-like ramification with axonal swellings in their course. In both types of mice at 2 weeks of age, comparatively thick nerve fibers with a smooth outline increased in number, and frequently ramified to form nerve terminals with dendritic profiles. However, no typical Ruffini endings with irregular outlines observed in the adult wild-type mice were found in the periodontal ligament at this stage. At postnatal 3 weeks, typical Ruffini endings with irregular outlines were discernable in the periodontal ligament of the wild-type mice while the dendritic endings showing smooth outlines were restricted to the homozygous mice. After postnatal 8 weeks, both types of mice showed an increase in the number of Ruffini endings, but the morphology differed between the wildtype and NT-4/5 homozygous mice. In the wild-type mice, a major population of the Ruffini endings expanded their axonal branches and developed many microprojections, resulting in a reduction of endings with smooth outlines. In contrast, we failed to find such typical Ruffini endings in the periodontal ligament of the homozygous mice: A majority of the periodontal Ruffini endings continued to show smooth outlines at postnatal 12 weeks. Quantitative analysis on neural density demonstrated a reduction in the homozygous mice with a significant difference by postnatal 8 weeks. Enzymatic histochemistry for non-specific ChE did not exhibit a distinct difference in the distribution and density of terminal Schwann cells between wild-type and homozygous mice. Furthermore, TrkB and p75-NGFR mRNA and proteins did not differ in the trigeminal ganglion between the two types. The periodontal Ruffini endings also displayed immunoreactivities for TrkB and p75-NGFR in both phenotypes. These findings suggest that the nt-4/5 gene depletion caused a delay in the formation and maturation of the periodontal Ruffini endings in the mice by inhibiting the growth of the periodontal nerves at an early stage, and indicate that multiple neurotrophins such as NT-4/5 and BDNF might play roles in the development and/or maturation of the periodontal Ruffini endings.

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  • Histological evidence of the altered distribution of osteocytes and bone matrix synthesis in klotho-deficient mice 査読

    H Suzuki, N Amizuka, K Oda, MQ Li, H Yoshie, H Ohshima, M Noda, T Maeda

    ARCHIVES OF HISTOLOGY AND CYTOLOGY   68 ( 5 )   371 - 381   2005年12月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:INT SOC HISTOLOGY & CYTOLOGY  

    Mice homozygous for klotho gene deletion are well established aging models as they mimic certain aspects of human senescence e.g. osteoporosis. Induced senescence may affect cellular functions and alter the histological properties of the extracellular matrices. The present study examined the histological and ultrastructural features of osteocytes and the surrounding bone matrix in klotho-deficient mice. As expected, osteoblasts showed a flattened shape with a weak immunoreactivity for alkaline phosphatase, and the bone matrix contained many empty osteocytic lacunae. The walls of both normal and empty lacunae were intensely immunopositive for osteopontin and dentin matrix protein-1, but featured an inconsistent immunoreactivity for osteocalcin and type I collagen. Not surprisingly, TUNEL-positivity, indicative of apoptosis, was found in many osteoblasts, osteocytes, and bone marrow cells of the klotho-deficient mice. In transmission electron microscopy, an amorphous matrix containing non-collagenous organic materials was recognizable around osteoblasts and in the osteocytic lacunae. Some osteoblasts on the bone surface featured these amorphous materials in vacuoles associated with their trans-Golgi network, indicating that, under klotho-deficient conditions, they synthesize and secrete the non-collagenous structures. Some osteocytes displayed pyknosis or degenerative traits. Thus, our findings provide histological evidence that klotho gene deletion influences the spatial distribution of osteocytes and the synthesis of bone matrix proteins in addition to the accelerated aging of bone cells.

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  • Postnatal development of substance P-immunoreaction in the trigerninal caudalis of neonatally capsaicin-treated mice 査読

    M Aita, T Maeda, R Takagi, K Seo

    ARCHIVES OF HISTOLOGY AND CYTOLOGY   68 ( 4 )   311 - 320   2005年12月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:INT SOC HISTOLOGY & CYTOLOGY  

    The trigeminal subnucleus caudalis (Vc) is a critical relay site for processing nociceptive afferent input from the orofacial area in addition to its modulation by neuroplastic change. Although an administration of capsaicin in neonates induces a selective destruction of substance P (SP)-immunoreactive nerve fibers, little information is available regarding its detailed effects on the Vc, particularly during postnatal development. The present study examined postnatal changes in the distribution of SP in the Vc and trigeminal ganglion (TG) by immunohistochemical techniques in naive (NV) and neonatally capsaicin-treated (CP) mice, combined with a quantitative analysis. The neonatal mice received a single subcutaneous injection of capsaicin (50 mg/kg) at 48 hours after birth. The neural density of the SP-immunoreaction decreased to approximately a quarter of that in 1-week-old NV mice but increased to three-quarters of that in the NV in the superficial area after postnatal week 2. A double staining with SP and myelin basic protein confirmed the absence of any SP-immunoreaction in the myelinated nerve fibers in both NV and CP mice. The SP-immunoreaction never overlapped with non-peptidergic IB4-labeled neurons in the Vc and TG of either group. Neither the size distribution of SP-positive neurons nor their relative ratio in the TG differed between NV and CP mice at the ages of postnatal weeks 1 and 8. These findings indicate two putative origins for the emergent SP-immunoreaction in the superficial layer of the Vc of the CP mice: the surviving trigeminal neurons with SP against capsaicin treatment and/or intrinsic neurons/interneurons in the Vc without SP under normal conditions.

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  • Postnatal development of excitation propagation in the trigeminal subnucleus caudalis evoked by afferent Stimulation in mice 査読

    Aita M, Seo K, Fujiwara N, Rituo T, Maeda T

    Developmental Brain Res   155   33 - 41   2005年12月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

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  • A histological evaluation for guided bone regeneration induced by a collagenous membrane 査読

    Y Taguchi, N Amizuka, M Nakadate, H Ohnishi, N Fujii, K Oda, S Nomura, T Maeda

    BIOMATERIALS   26 ( 31 )   6158 - 6166   2005年11月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:ELSEVIER SCI LTD  

    This study was designed to evaluate the histological changes during ossification and cellular events including osteogenic differentiation responding to collagenous bioresorbable membranes utilized for GBR. Standardized artificial bony defects were prepared at rat maxillae, and covered with a collagenous bioresorbable membrane. These animals were sacrificed at 1, 2, 3 and 4 weeks after the GBR-operation. The paraffin sections were subject to tartrate resistant acid phosphatase (TRAP) enzyme histochemistry and immunohistochemistry for alkaline phosphatase (ALP), osteopontin (OP) and osteocalcin (OC). In the first week of the experimental group, woven bone with ALP-positive osteoblasts occupied the lower half of the cavity. The collagenous membrane included numerous ALP-negative cells and OP-immunoreactive extracellular matrices. At 2 weeks, the ALP-, OP- and OC-immunoreactivity came to be recognizable in the region of collagenous membrane. Since ALP-negative soft tissue separated the collagenous membrane and the new bone originating from the cavity bottom, the collagenous membrane appeared to induce osteogenesis in situ. At 3 weeks, numerous collagen fibers of the membrane were embedded in the adjacent bone matrix. At 4 weeks, the membrane-associated and the cavity-derived bones had completely integrated, showing the same height of the periosteal ridge as the surrounding alveolar bones. The collagen fibers of a GBR-membrane appear to participate in osteogenic differentiation. (c) 2005 Elsevier Ltd. All rights reserved.

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  • Development of the articular cavity in the rat temporomandibular joint with special reference to the behavior of endothelial cells and macrophages 査読

    Akiko Suzuki, Kayoko Nozawa-Inoue, Nobuyuki Ikeda, Norio Amizuka, Kazuhiro Ono, Ritsuo Takagi, Takeyasu Maeda

    Anatomical Record - Part A Discoveries in Molecular, Cellular, and Evolutionary Biology   286 ( 2 )   908 - 916   2005年10月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Previous developmental studies on the temporomandibular joint (TMJ) have proposed several hypotheses on the formation of its articular cavity. However, detailed information is meager. The present study examined the formation process of the articular cavity in the rat TMJ by immunocytochemistry for CD31, RECA-1, and ED1, which are useful cellular markers for endothelial cells and monocyte/macrophage lineages, respectively. The upper articular cavity formation had begun by embryonic day 21 (E21) and was completed at postnatal day 1 (P1) in advance of the lower cavitation
    the latter took place from P1 to P3. The occurrence and distribution pattern of the CD31-, RECA-1-, and ED1-positive cells differed between the upper and lower articular cavity-forming areas: the ED1-positive cells exclusively occurred in the area of the prospective upper articular cavity prior to its formation, while no ED1-positive cell appeared in the lower cavity-forming area. In contrast, the CD31- and RECA-1-positive endothelial cells were restricted to the lower cavity-forming area (never the prospective upper cavity) at E19 and diminished thereafter. Throughout the cavity formation, we failed to find any apoptotic cells in the cavity formation area, indicating no involvement of apoptosis in the cavity formation in TMJ. The present findings on the behaviors of endothelial cells and ED1-positive cells show a possibility of different mechanism in the cavity formation between the upper and lower articular cavities in the rat TMJ. The appearance of ED1-reactive cells and temporal vascularization may play crucial roles in the upper and lower articular cavity formation, respectively. © 2005 Wiley-Liss, Inc.

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  • [Calcitonin receptor gene recombinant animals]. 査読

    Suzuki H, Amizuka N, Maeda T

    Nihon rinsho. Japanese journal of clinical medicine   63 Suppl 10   188 - 193   2005年10月

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  • Development of the articular cavity in the rat temporomandibular joint with special reference to the behavior of endothelial cells and macrophages 査読

    A Suzuki, K Nozawa-Inoue, N Ikeda, N Amizuka, K Ono, R Takagi, T Maeda

    ANATOMICAL RECORD PART A-DISCOVERIES IN MOLECULAR CELLULAR AND EVOLUTIONARY BIOLOGY   286A ( 2 )   908 - 916   2005年10月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:WILEY-LISS  

    Previous developmental studies on the temporomandibular joint (TMJ) have proposed several hypotheses on the formation of its articular cavity. However, detailed information is meager. The present study examined the formation process of the articular cavity in the rat TMJ by immunocytochemistry for CD31, RECA-1, and ED1, which are useful cellular markers for endothelial cells and monocyte/macrophage lineages, respectively. The upper articular cavity formation had begun by embryonic day 21 (E21) and was completed at postnatal day 1 (P1) in advance of the lower cavitation; the latter took place from P1 to P3. The occurrence and distribution pattern of the CD31-, RECA-1-, and ED1-positive cells differed between the upper and lower articular cavity-forming areas: the ED1-positive cells exclusively occurred in the area of the prospective upper articular cavity prior to its formation, while no ED1-positive cell appeared in the lower cavity-forming area. In contrast, the CD31- and RECA-1-positive endothelial cells were restricted to the lower cavity-forming area (never the prospective upper cavity) at E19 and diminished thereafter. Throughout the cavity formation, we failed to find any apoptotic cells in the cavity formation area, indicating no involvement of apoptosis in the cavity formation in TMJ. The present findings on the behaviors of endothelial cells and ED1-positive cells show a possibility of different mechanism in the cavity formation between the upper and lower articular cavities in the rat TMJ. The appearance of ED1-reactive cells and temporal vascularization may play crucial roles in the upper and lower articular cavity formation, respectively. (C) 2005 Wiley-Liss, Inc.

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  • ペリオスチン遺伝子欠損マウスの歯の矯正移動初期における組織学的検索

    李 敏啓, 網塚 憲生, パウロエンリケ・ルイズデフレイタズ, 前田 健康

    Journal of Oral Biosciences   47 ( Suppl. )   105 - 105   2005年9月

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  • 顎関節滑膜におけるCD44の局在

    鈴木 晶子, 野澤 佳世子[井上], 網塚 憲生, 前田 健康

    Journal of Oral Biosciences   47 ( Suppl. )   102 - 102   2005年9月

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  • Blomstrand型およびJansen型軟骨異形成症を誘導する変異型PTH/PTHrP受容体の細胞内局在異常について

    下村 淳子, 網塚 憲生, 小島 拓, 前田 健康, 下岡 正八

    Journal of Oral Biosciences   47 ( Suppl. )   101 - 101   2005年9月

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    記述言語:日本語   出版者・発行元:(一社)歯科基礎医学会  

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  • ラット顎関節におけるカベオリン-1の局在

    野澤 佳世子[井上], 鈴木 晶子, 網塚 憲生, 前田 健康

    Journal of Oral Biosciences   47 ( Suppl. )   102 - 102   2005年9月

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    記述言語:日本語   出版者・発行元:(一社)歯科基礎医学会  

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  • インプラント周囲に充填したα-TCP系骨補填材が誘導する新生骨について

    中舘 正芳, 網塚 憲生, 野村 修一, 前田 健康

    Journal of Oral Biosciences   47 ( Suppl. )   100 - 100   2005年9月

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    記述言語:日本語   出版者・発行元:(一社)歯科基礎医学会  

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  • ポリ乳酸プレートとhydroxyapatite/atelocollagen複合骨補填材による骨増生の組織化学的検索

    小島 拓, 網塚 憲生, 鈴木 晶子, 芳澤 享子, 齊藤 力, 前田 健康

    Journal of Oral Biosciences   47 ( Suppl. )   100 - 100   2005年9月

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    記述言語:日本語   出版者・発行元:(一社)歯科基礎医学会  

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  • Histochemical evidences on the chronological alterations of the hypertrophic zone of mandibular condylar cartilage 査読

    KS Hossain, N Amizuka, N Ikeda, K Nozawa-Inoue, A Suzuki, MQ Li, K Takeuchi, M Aita, Y Kawano, M Hoshino, K Oda, R Takagi, T Maeda

    MICROSCOPY RESEARCH AND TECHNIQUE   67 ( 6 )   325 - 335   2005年8月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:WILEY-LISS  

    The hypertrophic chondrocytes lack the ability to proliferate, thus permitting matrix mineralization as well as vascular invasion from the bone in both the mandibular condyle and the epiphyseal cartilage. This study attempted to verify whether the histological appearance of the hypertrophic chondrocytes is in a steady state during postnatal development of the mouse mandibular condyle. Type X collagen immunohistochemistry apparently distinguished the fibrous layer described previously as the "articular zone," "articular layer," and "resting zone" from the hypertrophic zone. Interestingly, the ratio of the type X collagen-positive hypertrophic zone in the entire condyle seemed higher in the early stages but decreased in the later stages. Some apparently compacted cells in the hypertrophic zone showed proliferating cell nuclear antigen (PCNA) immunoreaction, indicating the potential for cell proliferation at the early stages. As the mice matured, in contrast, they further enlarged and assumed typical features of hypertrophic chondrocytes. Apoptotic cells were also discernible in the hypertrophic zone at the early but not later stages. Consistent with morphological configurations of hypertrophic chondrocytes, immunoreactions for alkaline phosphatase, osteopontin, and type I collagen were prominent at the later stage, but not the early stage. Cartilaginous matrices demonstrated scattered patches of mineralization at the early stage, but increased in their volume and connectivity at the later stage. Thus, the spatial and temporal occurrence of these immunoreactions as well as apoptosis likely reflect the prematurity of hypertrophying cells at the early stage, and imply a physiological relevance during the early development of the mandibular condyles.

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  • [The interplay of magnesium and vitamin K2 on bone mineralization]. 査読

    Amizuka N, Li M, Maeda T

    Clinical calcium   15 ( 7 )   57 - 61   2005年7月

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  • Postnatal development of excitation propagation in the trigeminal subnucleus caudalis evoked by afferent stimulation in mice 査読

    K Seo, N Fujiwara, K Takeuchi, T Maeda, G Someya

    NEUROSCIENCE RESEARCH   52 ( 3 )   201 - 210   2005年7月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:ELSEVIER IRELAND LTD  

    The postnatal development of nociceptive afferent activity expansion and its modulation features were examined in mice using an optical imaging technique. Developing mice (1-2 weeks old (N 1-2w), 3-4 weeks old (N3-4w), 5-6 weeks old (N5-6w) and 7-8 weeks old (N7-8w)) and neonatally capsaicin-treated mice were used. The propagation of neuronal excitation was measured by changes in fluorescent intensity in horizontal brain stem slices evoked by electrical stimulation to the trigeminal spinal tract. A single-pulse stimulation evoked excitation propagation in the trigeminal caudalis (Vc). The propagation area was larger in N1-2w than in N7-8w, and no differences were observed between capsaicin-treated and naive mice in the same age groups. Repetitive stimulation (100 Hz, 30 pulses) elicited long-lasting and widespread excitation propagation. The excitation propagation area was significantly larger in N7-8w than in N1-2w, N3-4w and N5-6w. This propagation was suppressed by 5 mu M L-703.606, an NK1-receptor antagonist, suggesting that the repetitive stimulation-elicited excitation may require substance-P releases. Morphological observations demonstrated that the neural network in the Vc had grown by postnatal week 5. These results suggest that nociceptive afferent activity co-operatively matures with development of the network structure in the Vc, and that a mechanism for prolonged increase in central excitability is established during a later postnatal period. (c) 2005 Elsevier Ireland Ltd and the Japan Neuroscience Society. All rights reserved.

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  • 【骨代謝疾患の成因と治療 基礎と臨床をつなぐもの】骨基質石灰化におけるマグネシウムとビタミンK2の相関

    網塚 憲生, 李 敏啓, 前田 健康

    Clinical Calcium   15 ( 7 )   1155 - 1159   2005年7月

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    記述言語:日本語   出版者・発行元:(株)医薬ジャーナル社  

    骨基質の石灰化ミネラルにはさまざまな元素が存在するが,特に体内のマグネシウムは半分以上が骨基質に存在しており,カルシウム,リンとともに重要な骨基質ミネラルを構成している.低マグネシウム環境では,破骨細胞の吸収活性が上昇するとともに骨芽細胞も活性化され,骨代謝回転が亢進する.一方,ビタミンK2(MK-4:menatetrenone)は破骨細胞の活性を抑制するために骨代謝回転の過剰な亢進を是正する.一方で,低マグネシウム環境では,骨基質内のカルシウム濃度が上昇するために,純度の高いハイドロキシアパタイト結晶を形成し,石灰化の過剰成長までも誘導してしまう.ビタミンK2はハイドロキシアパタイト結晶の純度には影響を与えないが,石灰化の過剰成長を抑制することが明らかとなった(著者抄録)

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  • Immunocytochemical localization of MAPKAPK-2 and Hsp25 in the rat temporomandibular joint 査読

    Kayoko Nozawa-Inoue, Norio Amizuka, Akiko Suzuki, Takeyasu Maeda

    Anatomical Record - Part A Discoveries in Molecular, Cellular, and Evolutionary Biology   284 ( 2 )   522 - 528   2005年6月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    One series of our research has shown an intense expression of immunoreaction for heat shock protein 25 (Hsp25) in various cellular elements in the rat temporomandibular joint (TMJ). This protein is the major substrate of mitogen-activated protein kinase-activated protein kinase-2 (MAPKAPK-2), which mediates an intracellular stress-activated signaling pathway to stimulate cytosolic actin reorganization under various stresses. The present study was undertaken to examine the localization of MAPKAPK-2 in the rat TMJ by immunocytochemical techniques. Furthermore, confocal microscopy with double staining was employed to demonstrate the colocalization of MAPKAPK-2 and Hsp25. Immunocytochemistry for MAPKAPK-2 showed an intense immunoreaction in the cytoplasm of the synovial lining cells, the endothelial cells, and the fibroblasts in the synovial membrane of the rat TMJ. Double immunostaining under a confocal microscope succeeded in demonstrating the colocalization of MAPKAPK-2 and Hsp25 immunoreactions in the cytoplasm of fibroblastic type B synoviocytes in the TMJ. On the other hand, the macrophage-like type B-cells expressed MAPKAPK-2 immunoreactions but lacked Hsp25 immunoreactivity. The cells in the articular disk and the chondrocytes in the maturative and hypertrophic layer of the mandibular cartilage also showed intense immunoreactions for MAPKAPK-2 and Hsp25. In addition to cytoplasmic localization, MAPKAPK-2 immunoreactions were found in the nucleus of some synovial lining cells, cells in the articular disk, and chondrocytes. Current observations imply the presence of the phosphorylation of Hsp25 via activated MAPKAPK-2 in the cytoplasm. MAPKAPK-2 and Hsp25 possibly participate in the induction of cytoskeletal changes to the various cellular elements in rat TMJ under normal conditions. © 2005 Wiley-Liss, Inc.

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  • Characteristic distribution of immunoreaction for estrogen receptor alpha in rat ameloblasts 査読

    VL Ferrer, T Maeda, Y Kawano

    ANATOMICAL RECORD PART A-DISCOVERIES IN MOLECULAR CELLULAR AND EVOLUTIONARY BIOLOGY   284A ( 2 )   529 - 536   2005年6月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:WILEY-LISS  

    Estrogen has a diverse function, including cell proliferation and differentiation via estrogen receptors (ER), which have been reported to be the case in various tissues in addition to female reproductive organs. A recent immunocytochemical study has reported the expression of ER&alpha;, a subtype of ER, in rat odontoblasts, suggesting an involvement of estrogen in the differentiation of tooth-forming cells. However, there is no information on the ER&alpha; immunoexpression in ameloblasts. The present study was therefore undertaken to examine the localization of ERa immunoreaction in rat ameloblasts during amelogenesis. A computer-assisted quantitative analysis under a confocal laser scanning microscope was employed to demonstrate the stage-specific localization pattern of ERa immunoreaction. Immunohistochemistry of the rat enamel organ revealed ERa expression as nuclear localization in ameloblasts, stratum intermedium, stellate reticulum, and papillary layer, in addition to mature and immature odontoblasts. The ratio of immunopositive nuclei to total nuclei (immunopositive ratio) in ameloblasts was high at the apical loop region and gradually declined at the presecretory stage to zero at the secretory stage with statistically significant difference. The ER&alpha; immunolabeling pattern exhibited a periodic change at the maturation stage proper with constant higher labeling in ruffle-ended ameloblasts than in smooth-ended ameloblasts. The positive ratio was then followed by a statistically significant increase in immunolabeling thereafter. This stage-specific immunolabeling pattern during amelogenesis suggests a possible role of ER&alpha; in ameloblast proliferation and differentiation. &COPY; 2005 Wiley-Liss, Inc.

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  • Characteristic distribution of immunoreaction for estrogen receptor alpha in rat ameloblasts 査読

    Vesna-Lea Ferrer, Takeyasu Maeda, Yoshiro Kawano

    Anatomical Record - Part A Discoveries in Molecular, Cellular, and Evolutionary Biology   284 ( 2 )   529 - 536   2005年6月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Estrogen has a diverse function, including cell proliferation and differentiation via estrogen receptors (ER), which have been reported to be the case in various tissues in addition to female reproductive organs. A recent immunocytochemical study has reported the expression of ERα, a subtype of ER, in rat odontoblasts, suggesting an involvement of estrogen in the differentiation of tooth-forming cells. However, there is no information on the ERα immunoexpression in ameloblasts. The present study was therefore undertaken to examine the localization of ERα immunoreaction in rat ameloblasts during amelogenesis. A computer-assisted quantitative analysis under a confocal laser scanning microscope was employed to demonstrate the stage-specific localization pattern of ERα immunoreaction. Immunohistochemistry of the rat enamel organ revealed ERα expression as nuclear localization in ameloblasts, stratum intermedium, stellate reticulum, and papillary layer, in addition to mature and immature odontoblasts. The ratio of immunopositive nuclei to total nuclei (immunopositive ratio) in ameloblasts was high at the apical loop region and gradually declined at the presecretory stage to zero at the secretory stage with statistically significant difference. The ERα immunolabeling pattern exhibited a periodic change at the maturation stage proper with constant higher labeling in ruffle-ended ameloblasts than in smooth-ended ameloblasts. The positive ratio was then followed by a statistically significant increase in immunolabeling thereafter. This stage-specific immunolabeling pattern during amelogenesis suggests a possible role of ERα in ameloblast proliferation and differentiation. © 2005 Wiley-Liss, Inc.

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  • Immunocytochemical localization of MAPKAPK-2 and Hsp25 in the rat temporomandibular joint 査読

    K Nozawa-Inoue, N Amizuka, A Suzuki, T Maeda

    ANATOMICAL RECORD PART A-DISCOVERIES IN MOLECULAR CELLULAR AND EVOLUTIONARY BIOLOGY   284A ( 2 )   522 - 528   2005年6月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:WILEY-LISS  

    One series of our research has shown an intense expression of immuno-reaction for heat shock protein 25 (Hsp25) in various cellular elements in the rat temporomandibular joint (TMJ). This protein is the major substrate of mitogen-activated protein kinase-activated protein kinase-2 (MAPKAPK-2), which mediates an intracellular stress-activated signaling pathway to stimulate cytosolic actin reorganization under various stresses. The present study was undertaken to examine the localization of MAPKAPK-2 in the rat TMJ by immunocytochemical techniques. Furthermore, confocal microscopy with double staining was employed to demonstrate the colocalization of MAPKAPK-2 and Hsp25. Immunocytochemistry for MAPKAPK-2 showed an intense immunoreaction in the cytoplasm of the synovial lining cells, the endothelial cells, and the fibroblasts in the synovial membrane of the rat TMJ. Double immunostaining under a confocal microscope succeeded in demonstrating the colocalization of MAPKAPK-2 and Hsp25 immunoreactions in the cytoplasm of fibroblastic type B synoviocytes in the TMJ. On the other hand, the macrophage-like type A-cells expressed MAPKAPK-2 immunoreactions but lacked Hsp25 immunoreactivity. The cells in the articular disk and the chondrocytes in the maturative and hypertrophic layer of the mandibular cartilage also showed intense immunoreactions for MAPKAPK-2 and Hsp25. In addition to cytoplasmic localization, MAPKAPK-2 immunoreactions were found in the nucleus of some synovial lining cells, cells in the articular disk, and chondrocytes. Current observations imply the presence of the phosphorylation of Hsp25 via activated MAPKAPK-2 in the cytoplasm. MAPKAPK-2 and Hsp25 possibly participate in the induction of cytoskeletal changes to the various cellular elements in rat TMJ under normal conditions. &COPY; 2005 Wiley-Liss, Inc.

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  • チタンインプラント植立時に併用したα-TCP系自己硬化型補填材が新生骨へ及ぼす影響

    中舘 正芳, 網塚 憲生, 野村 修一, 前田 健康

    日本骨代謝学会学術集会プログラム抄録集   23回   220 - 220   2005年6月

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    記述言語:日本語   出版者・発行元:(一社)日本骨代謝学会  

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  • 肺癌骨転移巣における破骨細胞の骨吸収及び基質分解の形態学的観察

    李 敏啓, 網塚 憲生, 前田 健康

    日本骨形態計測学会雑誌   15 ( 2 )   77 - 77   2005年6月

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    記述言語:日本語   出版者・発行元:日本骨形態計測学会  

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  • ポリ乳酸プレートとハイドロキシアパタイト骨補填材を併用した骨増生の試み

    小島 拓, 網塚 憲生, 鈴木 晶子, 芳澤 享子, 齊藤 力, 前田 健康

    日本骨形態計測学会雑誌   15 ( 2 )   86 - 86   2005年6月

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    記述言語:日本語   出版者・発行元:日本骨形態計測学会  

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  • ポリ乳酸プレートとハイドロキシアパタイト骨補填材を応用した骨増生法の開発

    小島 拓, 網塚 憲生, 鈴木 晶子, 芳澤 享子, 齊藤 力, 前田 健康

    日本骨代謝学会学術集会プログラム抄録集   23回   236 - 236   2005年6月

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    記述言語:日本語   出版者・発行元:(一社)日本骨代謝学会  

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  • 骨基質の石灰化過程におけるビタミンKの微細構造学的作用について

    網塚 憲生, 李 敏啓, 原 久仁子, 小林 正敏, 竹内 亀一, 秋山 康博, 前田 健康

    日本骨代謝学会学術集会プログラム抄録集   23回   274 - 274   2005年6月

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    記述言語:日本語   出版者・発行元:(一社)日本骨代謝学会  

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  • 培養骨芽細胞における変異型PTH/PTHrP受容体の細胞内局在異常について

    下村 淳子, 網塚 憲生, 小島 拓, 前田 健康, 下岡 正八

    日本骨代謝学会学術集会プログラム抄録集   23回   174 - 174   2005年6月

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    記述言語:日本語   出版者・発行元:(一社)日本骨代謝学会  

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  • インプラント周囲に充填した自己硬化型α-TCP系骨補填材に対する組織反応

    中舘 正芳, 魚島 勝美, 網塚 憲生, 織田 公光, 前田 健康, 野村 修一

    日本補綴歯科学会雑誌   49 ( 113回特別 )   74 - 74   2005年5月

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    記述言語:日本語   出版者・発行元:(公社)日本補綴歯科学会  

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  • Postnatal changes in the spatial distributions of substance P and neurokinin-1 receptor in the trigeminal subnucleus caudalis of mice 査読

    M Aita, K Seo, N Fujiwara, R Takagi, T Maeda

    DEVELOPMENTAL BRAIN RESEARCH   155 ( 1 )   33 - 41   2005年3月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:ELSEVIER SCIENCE BV  

    Nociceptive afferent signals from the orofacial area are transmitted to the trigeminal subnucleus caudalis (Vc) through the release of glutamate and/or substance P (SP). Although nociceptive transmission and/or modulating mechanisms are known to develop during the postnatal period, the specific developmental changes in nociception and/or modulation remain unclear. The present study examined postnatal changes in the spatial relationship between SP and its receptor, the NK1 receptor (NK1R), in the mouse Vc by immunohistochemistry and quantitative analysis. The medulla was removed from C57BL/6N mice (1, 2, 4, and 8 weeks of age) after perfusion and fixation, and cut horizontally at a thickness of 40 mu m. The relative densities of SP- and NK1R-immunoreactive areas and their changes with age were assessed statistically. One- and 2-week-old mice showed relatively high densities of SP-positive structures in the marginal layer (Mar) and the deep part of the magnocellular layer (Mag). The SP distribution in the superficial Vc remained unchanged, but the density in the deep Mag gradually decreased with age, resulting in a complete loss after postnatal week 4. The NK1R-irnmunoreactivity exhibited a similar distribution pattern to that of SP, but the pattern remained unchanged during the postnatal period.
    Double-iinrnunofluorescence staining for SP and NK1 R demonstrated only moderate direct contact of SP-positive structures with NK1R in the superficial area. These separate distributions and the postnatal changes in SP and NK1R suggest the possibility of another nociceptive afferent transmission mechanists, that is, volume transmission, in the Vc other than synapse-mediated transmission. (c) 2005 Elsevier B.V All rights reserved.

    DOI: 10.1016/j.devbrainres.2004.12.005

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  • 骨吸収型骨転移病巣で破骨細胞が産生するcathepsin KとMMP-9の免疫局在

    李 敏啓, 網塚 憲生, 井上 佳世子, 河野 芳郎, 竹内 亀一, 前田 健康

    解剖学雑誌   80 ( Suppl. )   161 - 161   2005年3月

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    記述言語:日本語   出版者・発行元:(一社)日本解剖学会  

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  • op/opマウスにおける骨基質石灰化と骨芽細胞の局在について

    坂上 直子, 網塚 憲生, 李 敏啓, 宇田川 信之, 前田 健康

    解剖学雑誌   80 ( Suppl. )   143 - 143   2005年3月

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    記述言語:日本語   出版者・発行元:(一社)日本解剖学会  

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  • 致死型軟骨無形成症の組織異常と細胞内シグナリング 小胞体からのアポトーシスシグナルの可能性

    那須 真樹子, 網塚 憲生, 李 敏啓, 野村 修一, 前田 健康

    新潟歯学会雑誌   34 ( 2 )   243 - 246   2005年1月

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    記述言語:日本語   出版者・発行元:新潟歯学会  

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  • 肺癌骨転移病巣において破骨細胞が産生するcathepsin KとMMP-9の局在

    李 敏啓, 網塚 憲生, 織田 公光, 河野 芳郎, 竹内 亀一, 前田 健康

    新潟歯学会雑誌   34 ( 2 )   287 - 287   2005年1月

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    記述言語:日本語   出版者・発行元:新潟歯学会  

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  • 低マグネシウムラットにおける骨代謝および石灰化に関する微細構造学的検討

    李 敏啓, 網塚 憲生, 竹内 亀一, 前田 健康

    新潟歯学会雑誌   34 ( 2 )   288 - 288   2005年1月

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    記述言語:日本語   出版者・発行元:新潟歯学会  

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  • 変異型線維芽細胞増殖因子受容体III型による軟骨細胞のアポトーシス

    那須 真樹子, 網塚 憲生, 李 敏啓, 野村 修一, 前田 健康

    新潟歯学会雑誌   34 ( 2 )   286 - 286   2005年1月

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    記述言語:日本語   出版者・発行元:新潟歯学会  

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  • 致死型軟骨無形成症II型における骨端軟骨への血管侵入異常

    那須 真樹子, 網塚 憲生, 李 敏啓, 野村 修一, 前田 健康

    新潟歯学会雑誌   34 ( 2 )   287 - 287   2005年1月

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    記述言語:日本語   出版者・発行元:新潟歯学会  

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  • Histological evaluation for "bone quality" on two mouse models with different bone remodeling 査読

    N Amizuka, J Shimomura, MQ Li, M Nasu, T Maeda

    JOURNAL OF BONE AND MINERAL METABOLISM   23   43 - 47   2005年

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:SPRINGER TOKYO  

    DOI: 10.1007/BF03026322

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  • Reduced osteoblastic population and defective mineralization in osteopetrotic (op/op) mice 査読

    N Sakagami, N Amizuka, MQ Li, K Takeuchi, M Hoshino, M Nakamura, K Nozawa-Inoue, N Udagawa, T Maeda

    MICRON   36 ( 7-8 )   688 - 695   2005年

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    記述言語:英語   出版者・発行元:PERGAMON-ELSEVIER SCIENCE LTD  

    Osteopetrotic (op/op) mice fail to exhibit bone remodeling because of a defective osteoclast formation due to a lack of macrophage colony-stimulating factor. In this study, we investigated the femora of op/op mice to clarify whether the osteoblastic population and bone mineralization are involved in osteoclasts or their bone resorption. The op/op mice extended the meshwork of trabecular bones from the chondro-osseous junction to the diaphyseal region. In the femoral metaphyses of op/op mice, intense alkaline phosphatase (ALPase)-positive osteoblasts were observed on the metaphyseal bone in close proximity to the erosion zone of the growth plates. Von Kossa's staining revealed scattered mineralized nodules and a fine meshwork of mineralized bone matrices while the wild-type littermates developed well-mineralized trabeculae parallel to the longitudinal axis. In contrast to the metaphysis, some op/op diaphyses showed flattened osteoblasts with weak ALPase-positivity, and the other diaphyses displayed bone surfaces without a covering by osteoblasts. It is likely, therefore, that the osteoblastic population and activity were lessened in the op/op diaphyses. Despite the osteopetrotic model, von Kossa's staining demonstrated patchy unmineralized areas in the op/op diaphyses, indicating that a lower population and/or the activity of osteoblasts resulted in defective mineralization in the bone. Transmission electron microscopy disclosed few osteoblasts on the diaphyseal bones, and instead, bone marrow cells and vascular endothelial cells were often attached to the unmineralized bone. Osteocytes were embedded in the unmineralized bone matrix. Thus, osteoclasts appear to be involved in the osteoblastic population and activity as well as subsequent bone mineralization. (c) 2005 Elsevier Ltd. All rights reserved.

    DOI: 10.1016/j.micron.2005.06.008

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  • Ultrastructural Images of Enamel Tufts in Human Permanent Teeth

    Amizuka Norio, Uchida Takashi, Nozawa-Inoue Kayoko, Kawano Yoshiro, Suzuki Akiko, Li Minqi, Nasu Makiko, Kojima Taku, Sakagami Naoko, Ozawa Hidehiro, Maeda Takeyasu

    Journal of Oral Biosciences   47 ( 1 )   33 - 41   2005年

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    記述言語:英語   出版者・発行元:Japanese Association for Oral Biology  

    Human enamel tufts appeared as corrugated ribbon-like structures located on the dentin parallel to the tooth axis when observed under the binocular microscope and scanning electron microscope (SEM). SEM observation disclosed enamel tufts as bundles of well-extended tubular structures attributable to hypomineralized enamel sheaths. Plate-like structures, previously referred to as "tuft-root" ran in the center of the enamel tufts, connecting the dentin surface. When observing under the transmission electron microscope, the plates of tufts extended from the superficial layer of the dentin, penetrating the hypermineralized zone adjacent to the dentin-enamel (D-E) junction, and then, reaching the tuft region. In the tuft region, the plates of tufts ran mainly along the enamel sheaths and partially across the enamel prisms. The immuno-gold technique verified an intense immunoreactivity for amelogenin in the superficial layer of the dentin as well as the enamel prisms in the tufts, although no reaction was found over the "plates of tufts". The immunoreactivity for 13-17 kd sheath proteins, also denoted as sheathlin, ameloblastin or amelin, was detected over the filamentous structures closely associated with the enamel sheaths in the enamel tuft. Thus, our study demonstrated that enamel tufts consist of both well extended hypomineralized enamel prisms and "plates of tufts". The major organic substance of the enamel tufts is suggested to be 13-17 kd sheath proteins rather than amelogenin.

    DOI: 10.2330/joralbiosci.47.33

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  • 歯学教育プログラムへのPBL教育の導入−南カリフォルニア大学歯学部における実態調査− 査読

    安島久雄, 小野和宏, 前田健康, 永田昌毅, 高木律男, 山田好秋, 興地隆史, 魚島勝美, 葭原明弘, 花田晃治

    日歯教誌   10 ( 1 )   166 - 173   2004年12月

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    記述言語:日本語  

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  • Immunohistochemical localization of periostin in tooth and its surrounding tissues in mouse mandibles during development. 査読 国際誌

    Suzuki H, Amizuka N, Kii I, Kawano Y, Nozawa-Inoue K, Suzuki A, Yoshie H, Kudo A, Maeda T

    The anatomical record. Part A, Discoveries in molecular, cellular, and evolutionary biology   281 ( 2 )   1264 - 1275   2004年12月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    DOI: 10.1002/ar.a.20080

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  • GFAP immunoreactivity and transcription in trigeminal and dental tissues of rats and transgenic GFP/GFAP mice 査読

    MR Byers, A Maeda, AM Brown, RE Westenbroek

    MICROSCOPY RESEARCH AND TECHNIQUE   65 ( 6 )   295 - 307   2004年12月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:WILEY-LISS  

    Sensory mechanisms in teeth are not well understood and may involve pulpal-neural interactions. Tooth cells that proliferate in vitro have polyclonal immunoreactivity (IR) for glial fibrillary acidic protein, (GFAP), growth-associated protein (GAP-43), And vimentin, plus glial-like ion channels. Here, we analyzed GFAP-IR patterns in dental and trigeminal tissues of rats, for comparison With green fluorescent protein (GFP) associated with GFAP transcription in transgenic mice, in order to better characterize glial-like cells in dental tissues. Astrocytes, ganglion satellite cells, And epineurial Schwann cells were demonstrated by anti-GFAP antibodies and GFP-GFAP, as expected. Odontoblasts did not, stain by any of these methods and cannot be the glial-like cells. Fibroblasts and undifferentiated mesenchymal cells in pulp had polyclonal GFAP-IR and vimentin-IR, while nerve fibers reacted only with polyclonal antibody. Some Schwann cell subtypes in trigeminal nerve and Oral mucosa were positive for GFP and for polyclonal anti-GFAP, but not for monoclonal antibody. In pulp. almost all Schwann cells were unstained, but many Schwann cells in periodontal ligament had polyclonal GFAP-IR. These results show greater heterogeneity for Schwann cells than expected, and suggest that the glial-like pulp cells are fibroblasts and/or undifferentiated mesenchymal cells or stem cells. We also found that polyclonal GFAP revealed intermediate filaments in preterminal sensory nerve fibers, thereby providing a useful marker for that neural subregion. GFP transcription by some Schwann cell subtypes in oral mucosae and trigeminal nerve, but not trigeminal root Was a novel finding that reveals more complexity in peripheral glia than previously recognized. (C) 2005 Wiley-Liss, Inc.

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  • 発生期マウス下顎関節突起における肥大層の組織化学的変化(Histochemical changes of the hypertrophic zone in developing mouse mandibular condyle)

    Hossain Kazi Sazzad, 池田 順行, 野澤 佳世子[井上], 網塚 憲生, 織田 公光, 高木 律男, 前田 健康

    Journal of Oral Biosciences   46 ( 5 )   396 - 396   2004年9月

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    記述言語:英語   出版者・発行元:(一社)歯科基礎医学会  

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  • 破骨細胞が産生するcathepsin KとMMP-9の局在について 肺癌骨転移モデルを用いた検討

    李 敏啓, 網塚 憲生, 織田 公光, 河野 芳郎, 竹内 亀一, 那須 真樹子, 小島 拓, 高木 律男, 前田 健康

    Journal of Oral Biosciences   46 ( 5 )   405 - 405   2004年9月

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    記述言語:日本語   出版者・発行元:(一社)歯科基礎医学会  

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  • 肺癌骨転移モデルにおいて破骨細胞が産生する基質分解酵素の組織化学的検討

    李 敏啓, 網塚 憲生, 織田 公光, 河野 芳朗, 竹内 亀一, 高木 律男, 前田 健康

    日本骨代謝学会学術集会プログラム抄録集   22回   201 - 201   2004年8月

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    記述言語:日本語   出版者・発行元:(一社)日本骨代謝学会  

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  • klotho欠損マウスの象牙芽細胞と骨細胞における組織学的異常について

    鈴木 啓展, 網塚 憲生, 織田 公光, 野田 政樹, 吉江 弘正, 前田 健康

    日本骨代謝学会学術集会プログラム抄録集   22回   202 - 202   2004年8月

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    記述言語:日本語   出版者・発行元:(一社)日本骨代謝学会  

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  • Development of the synovial membrane in the rat temporomandibular joint as demonstrated by immunocytochemistry for heat shock protein 25. 査読

    Ikeda N, Nozawa-Inoue K, Takagi R, Maeda T

    The anatomical record. Part A, Discoveries in molecular, cellular, and evolutionary biology   279 ( 1 )   623 - 635   2004年7月

  • Development of the synovial membrane in the rat temporomandibular joint as demonstrated by immunocytochemistry for heat shock protein 25 査読

    N Ikeda, K Nozawa-Inoue, R Takagi, T Maeda

    ANATOMICAL RECORD PART A-DISCOVERIES IN MOLECULAR CELLULAR AND EVOLUTIONARY BIOLOGY   279A ( 1 )   623 - 635   2004年7月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:WILEY-LISS  

    The synovial lining layer of the temporomandibular joint (TMJ) consists of macrophage-like type A cells and fibroblast-like type B cells. Until now, little information has been available on the development of the synovial membrane in TMJ. In the present study we examined the development of the synovial lining layer in the rat TMJ by light- and electron-microscopic immunocytochemistry for heat shock protein (Hsp) 25, which is a useful marker for type B cells. At embryonic day 19 (E19), a few Hsp25-positive cells first appeared in the upper portion of the developing condyle. During the formation of the upper articular cavity (E21 to postnatal day 1 (P1)), a few positive cells were arranged on its surface. Immunoelectron microscopy demonstrated that these cells had ultrastructural features of fibroblast-like type B cells. In addition, some Hsp25-positive cells moved to the deep portion by extending their cytoplasmic processes toward the articular cavity at P3. At that time, the presence of typical macrophage-like type A cells in the lining layer was confirmed by immunoelectron microscopy. The slender processes of Hsp25-positive cells showed a continuous covering with the synovial surface at P7, followed by a drastic increase in the Hsp25-positive cells at P15 and later, when active jaw movement occurred. These findings suggested that the arrangement and morphological maturation of type B cells are closely related to the formation of the articular cavity in the embryonic period and the commencement of active jaw movement after birth, respectively. (C) 2004 Wiley-Liss, Inc.

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  • The involvement of brain-derived neurotrophic factor in the pattern generator of mastication. 査読

    Okayasu I, Yamada Y, Maeda T, Yoshida N, Koga Y, Oi K

    Brain research   1016 ( 1 )   40 - 47   2004年7月

  • A histological evaluation of the involvement of Bio-Oss (R) in osteoblastic differentiation and matrix synthesis 査読

    FI Tapety, N Amizuka, K Uoshima, S Nomura, T Maeda

    CLINICAL ORAL IMPLANTS RESEARCH   15 ( 3 )   315 - 324   2004年6月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:BLACKWELL MUNKSGAARD  

    This study was designed to investigate the responses of bone cells to a deproteinized bovine bone material, Bio-Oss(R) (Geistlich-Pharma, Wolhunsen, Switzerland), which was grafted in artificial bone defects of rat femurs. Standardized bone defects in the cortical bone of the right femurs were grafted with Bio-Oss(R) particles. Narrow penetrations were prepared on the bottom of the cavity, enabling osteogenic cells to migrate from the bone marrow. A defect in the left femur without Bio-Oss(R) was used as a control. The treated femurs were histochemically examined at 1, 3, 5, 7, and 14 days after the operation. At day 1, no osteogenic migration into the cavities occurred in either the control or experimental groups. At day 3, alkaline phosphatase (ALPase) immunohistochemistry showed a migration of the positive cells at the bottom of the cavities of the experimental groups, but not in the control ones. At day 5, new bone formation was recognized at the bottom of the cavity of both groups. In the experimental group, ALPase-positive cells were localized on Bio-Oss(R) and/or on the thin bone matrix that covered this material. The superficial layer of Bio-Oss(R) underlying the newly formed bone exhibited osteocalcin immunoreactivity. Transmission electron microscopy revealed osteoblasts depositing bone matrices - including collagen fibers - on the surface of Bio-Oss(R). At days 7 and 14, woven bone occupied the previous cavities of both control and experimental groups, accompanied by osteoclasts. Thus, Bio-Oss(R) appears to serve as a scaffold for osteogenic cells as well as to promote osteoblastic differentiation and matrix synthesis.

    DOI: 10.1111/j.1600-0501.2004.01012.x

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  • An immunocytochemical study of pulpal responses to cavity preparation by laser ablation in rat molars by using antibodies to heat shock protein (Hsp) 25 and class II MHC antigen. 査読

    Suzuki T, Nomura S, Maeda T, Ohshima H

    Cell and tissue research   315 ( 3 )   311 - 319   2004年3月

  • A histological evaluation on self-setting alpha-tricalcium phosphate applied in the rat bone cavity 査読

    H Hao, N Amizuka, K Oda, N Fujii, H Ohnishi, A Okada, S Nomura, T Maeda

    BIOMATERIALS   25 ( 3 )   431 - 442   2004年2月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:ELSEVIER SCI LTD  

    This study aimed to elucidate the biological effects of a self-setting tricalcium phosphate bone substitute (BIOPEX(R)) applied in rat femoral cortical bone cavities. Narrow penetrations through the cavity and bone marrow were prepared to obtain cellular sources. In the experimental group at day 1, a thin cell layer intruded into a narrow space between the grafted BIOPEX(R) and the bottom of the cavity. From days 5 to 10, a range of tartrate-resistant acid phosphatase (TRAPase)-reactive osteoclasts accumulated on the surface of the BIOPEX(R) facing the bottom of the cavity, whilst many alkaline phosphatase (ALPase)-positive osteoblasts were localized on the bone surface opposing the BIOPEX(R). However, at day 20, osteoblasts were localized neighboring the osteoclasts on the BIOPEX(R), and deposited bone matrices onto this material, implying a coupling between osteoclasts and osteoblasts. At days 30 and 40 post-operation, small remnants of BIOPEX(R) particles were present in the new bone with a profile of compact bone. Thus, BIOPEX(R) is resorbed by osteoclasts, and succeeded by osteoblastic bone apposition with a coupling of osteoclasts and osteoblasts at the later stage. In conclusion, the use of BIOPEX(R) provides adequate bone regeneration with the profile of compact bone. (C) 2003 Elsevier Ltd. All rights reserved.

    DOI: 10.1016/S0142-9612(03)00550-7

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  • Immunolocalization of corticosteroid hormone receptors in the mechanoreceptors in rat oral tissues 査読

    Shoko Kinoshita-Kawano, Yoshiro Kawano, Tadashi Noda, Takeyasu Maeda

    BIOMEDICAL RESEARCH-TOKYO   25 ( 1 )   35 - 43   2004年2月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:BIOMEDICAL RESEARCH PRESS LTD  

    Previous studies have confirmed the presence of corticosteroid receptors including mineralocorticoid (MCR) and glucocorticoid receptors (GCR) in the glia cells of both central and peripheral nervous systems. However, no report has been offered as yet on the detailed localization of these receptors in the specialized Schwann cells associated with mechanoreceptors. Thus, the present study examined the immunolocalization of MCR and GCR in the specialized Schwann cells (terminal and lamellar Schwann cells) associated with the mechanoreceptors in rat periodontal ligament and palatal mucosa by the use of immunohistochemical techniques and confocal laser scanning microscopy. It further attempted to detect the localization of 11 beta-hydroxysteroid dehydrogenase type 2 (11 beta-HSD II), a key enzyme for signal transduction via MCR. Immunostaining with antisera against GCR and MCR demonstrated intense immunoreactions in the terminal or lamellar Schwann cells but not nerve fibers associated with the periodontal and palatal mechanoreceptors. A double staining with propidium iodide and either GCR- or MCR-antiserum revealed the intranuclear and cytoplasmic localization of these receptors in the terminal Schwann cells. 11 beta-HSD II-immunoreactivity was found in the cytoplasm and on the nuclear envelope of the terminal Schwann cells, indicating the co-localization of MCR and 11 beta-HSD II in them. These findings suggest that the terminal or lamellar Schwann cells are target tissues for corticosteroid hormones. Taken together with previous reports on the functional significance of the corticosteroids, the results indicate that glucocorticoids and mineralocorticoids might be respectively involved in the proliferation and/or differentiation and Na+/K+-homeostasis in the terminal or lamellar Schwann cells associated with the mechanoreceptors.

    DOI: 10.2220/biomedres.25.35

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  • Signalling by fibroblast growth factor receptor 3 and parathyroid hormone-related peptide coordinate cartilage and bone development 査読

    N Amizuka, D Davidson, HL Liu, G Valverde-Franco, S Chai, T Maeda, H Ozawa, Hammond, V, DM Ornitz, D Goltzman, JE Henderson

    BONE   34 ( 1 )   13 - 25   2004年1月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:ELSEVIER SCIENCE INC  

    Bone development is regulated by conserved signalling pathways that are linked to multifunctional growth factors and their high affinity receptors. Parathyroid hormone-related peptide (PTHrP) and fibroblast growth factor receptor 3 (FGFR3) have been shown to play pivotal, and sometimes complementary, roles in the replication, maturation and death of chondrocytes during endochondral bone formation. To gain further insight into how these pathways coordinate cartilage and bone development, we generated mice lacking expression of both PTHrP and FGFR3. The phenotype of compound mutant mice resembled that of their PTHrP-deficient littermates with respect to neonatal lethality, facial dysmorphism and foreshortening of the limbs. The absence of PTHrP in the developing epiphyseal cartilage of PTHrP(- / -) and PTHrP(- / -) /FGFR3(- / -) mice resulted in a dominant hypo-proliferative phenotype. However, abnormalities such as the presence of nonhypertrophic cells among hypertrophic chondrocytes and excessive apoptosis seen in the hypertrophic zone of PTHrP(-) (/) (-) mice were absent in the PTHrP(- / -) /FGFR3(-) (/) (-) mice. Furthermore, the absence of FGFR3 in single and compound mutant mice led to decreased expression of vascular endothelial growth factor (VEGF) and an increase in depth of hypertrophic chondrocytes. These observations indicate that FGFR3 deficiency can rescue some of the defects seen in PTHrP-deficient mice and that it plays an important role in the regulation of chondrocyte differentiation and hypertrophy. These studies support a dominant role for PTHrP in regulating the pool of proliferating cells during limb development and suggest that signalling by FGFR3 plays a more prominent role in cartilage maturation and vascular invasion at the chondro-osseous junction. (C) 2003 Elsevier Inc. All rights reserved.

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  • 骨折の初期治癒過程における組織化学的検討

    李 敏啓, 網塚 憲生, 竹内 亀一, 高木 律男, 前田 健康

    新潟歯学会雑誌   33 ( 2 )   290 - 290   2004年1月

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    記述言語:日本語   出版者・発行元:新潟歯学会  

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  • ラット上顎骨における吸収性膜を用いたGBR法の組織学的観察

    田口 裕哉, 網塚 憲生, 中舘 正芳, 大西 英夫, 藤井 規孝, 野村 修一, 前田 健康

    新潟歯学会雑誌   33 ( 2 )   290 - 291   2004年1月

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    記述言語:日本語   出版者・発行元:新潟歯学会  

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  • 軟骨細胞においてFGFR3はJAK/STAT系を介してPTH/PTHrP受容体発現抑制を行う

    関 雪絵, 網塚 憲生, 永田 昌毅, 織田 公光, 高木 律男, 前田 健康

    新潟歯学会雑誌   33 ( 2 )   289 - 290   2004年1月

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    記述言語:日本語   出版者・発行元:新潟歯学会  

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  • 客観的臨床能力試験(OSCE)における概略評定の信頼性と妥当性−平成15年度新潟大学歯学部OSCEの解析− 査読

    興地隆史, 福島正義, 吉羽邦彦, 小林 博, 葭原明弘, 星名秀行, 富沢美恵子, 高木律男, 前田健康, 山田好秋

    日歯教誌   20 ( 1 )   135 - 141   2004年

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  • Malignant fibrous histiocytoma arising in abutting soft tissue after resection of a benign fibrous histiocytoma of the mandible 査読

    Kenji Izumi, Jun Cheng, Takeyasu Maeda, Takashi Saku

    Oral Oncology Extra   40 ( 2 )   24 - 28   2004年

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:Elsevier BV  

    We present a unique case of malignant fibrous histiocytoma (MFH) developing around a masseter muscle close to the surgical margin from which a benign fibrous histiocytoma (BFH) of the mandible was excised 3 years and 10 months earlier. In the interval between the surgical removal of the original BFH and the MFH, several surgical procedures were performed in the related region. Clinical and histopathological evidence showed that this case represented two separate lesions rather than a single disease process. The clinical history suggests that the chronic repair process might have contributed to the tumorigenic factor of a MFH. © 2004 Elsevier Ltd. All rights reserved.

    DOI: 10.1016/j.ooe.2003.12.001

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  • Synovial Type B Cells in the Temporomandibular Joint

    Nozawa-Inoue Kayoko, Ikeda Nobuyuki, Suzuki Akiko, Amizuka Norio, Maeda Takeyasu

    Journal of Oral Biosciences   46 ( 6 )   519 - 522   2004年

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    記述言語:英語   出版者・発行元:歯科基礎医学会  

    The synovial lining layer in the temporomandibular joint (TMJ) consists of macrophage-like type A cells and fibroblast-like type B cells. Their identification, however, has been difficult because of the lack of a specific cell marker. This review summarizes the characteristic configuration and development of the type B cell in the murine TMJ. Immunocytochemistry for 25kDa-heat shock protein (Hsp25) revealed two profiles of the fibroblast-like type B cells with cytoplasmic processes in the adult rat TMJ : one projected horizontally slender processes which covered the synovial membrane, and the other extended a thick, long process towards the articular cavity. The former appeared earlier than the latter during the development of synovial membrane, in close relation with the formation of the articular cavity and the commencement of active jaw movement. Since immature type B cells with Hsp25-immunoreactivity were found in the mesenchymal tissue which corresponded to the future articular cavity, type B cells may differentiate directly from mesenchymal cells.

    DOI: 10.2330/joralbiosci.46.519

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  • Tissue response to titanium implantation in the rat maxilla, with special reference to the effects of surface conditions on bone formation 査読

    M Shirakura, N Fujii, H Ohnishi, Y Taguchi, H Ohshima, S Nomura, T Maeda

    CLINICAL ORAL IMPLANTS RESEARCH   14 ( 6 )   687 - 696   2003年12月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:BLACKWELL MUNKSGAARD  

    Tissue responses to titanium implantation with two different surface conditions in our established implantation model in rat maxillae were investigated by light and transmission electron microscopy and by histochemistry for tartrate-resistant acid phosphatase (TRAPase) activity. Here we used two types of implants with different surface qualities: titanium implants sandblasted with Al2O3 (SA-group) and implants coated with hydroxyapatite (HA-group). In both groups, bone formation had begun by 5 days postimplantation when the inflammatory reaction had almost disappeared in the prepared bone cavity. In the SA-group, however, the bone formation process in the bone cavity was almost identical to that shown in our previous report using smooth surfaced implants (Futami et al. 2000): new bone formation, which occurred from the pre-existing bone toward the implant, was preceded by active bone resorption in the lateral area with a narrow gap, but not so in the base area with a wide gap. In the HA-group, direct bone formation from the implant toward the pre-existing bone was recognizable in both lateral and base areas. Many TRAPase-reactive cells were found near the implant surface. On the pre-existing bone, new bone formation occurred with bone resorption by typical osteoclasts. Osseointegration around the implants was achieved by postoperative day 28 in both SA- and HA-groups except for the lateral area, where the implant had been installed close to the cavity margin. These findings indicate that ossification around the titanium implants progresses in different patterns, probably dependent on surface properties and quality.

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  • Synovial membrane in the temporomandibular joint - Its morphology, function and development 査読

    K Nozawa-Inoue, N Amizuka, N Ikeda, A Suzuki, Y Kawano, T Maeda

    ARCHIVES OF HISTOLOGY AND CYTOLOGY   66 ( 4 )   289 - 306   2003年10月

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    記述言語:英語   出版者・発行元:INT SOC HISTOLOGY & CYTOLOGY  

    This paper reviews recent findings of the synovial membrane, in particular the morphology, function and development of synovial lining cells, in the temporomandibular joint (TMJ). Electron microscopic studies have confirmed the synovial membrane in TMJ consists of macrophage-like type A cells and fibroblast-like type B cells identical to those in other systematic joints. The macrophage-like type A cells react with anti-macrophage and macrophage-derived substances including the major histocompatibility class IT molecule, and show a drastic increase in their number in the inflamed synovial membrane. In addition, they have the ability to produce substances involved in the progression of TMJ inflammation such as nitric oxide and inducible nitric oxide synthase. Observation of osteopetrotic mice revealed that macrophage-like type A cells in TMJ are derived from monocyte lineage. Immunocytochemistry for 25kDa heat shock protein was able to depict the entire shape of fibro-blast-like type B cells including their unique processes. The expression of an estrogen receptor alpha-immunoreaction in the fibroblast-like type B cells may explain the etiology of temporomandibular disorders at a higher frequency in females than in males, suggesting that TMJ is a target tissue for estrogen. Furthermore, fibroblast-like type B cells are equipped with a basement membrane to serve as an adhesion molecule for the fibroblast-like type B cells to keep their epithelial arrangement. A clear understanding of the morphology of the intact synovial membrane will serve to clarify the etiology and development of temporomandibular disorders.

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  • Regional differences in the distribution and type of immunocompetent cells in the rat oral mucosae 査読

    A Suzuki, K Nozawa-Inoue, Y Kawano, N Amizuka, T Maeda

    BIOMEDICAL RESEARCH-TOKYO   24 ( 5 )   249 - 260   2003年10月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:BIOMEDICAL RESEARCH PRESS LTD  

    The distribution and type of immunocompetent cells were investigated in rat oral mucosae using immunocytochemistry and enzyme histochemistry, focusing on histological structures. We used two antibodies, OX6 and ED1, which recognize the rat la-antigen and macrophage/monocyte lineage, respectively. Enzymatic histochemistry for acid phosphatase (ACPase) activity and adenosine triphosphatase (ATPase) activity was also employed to identify macrophages and Langerhans cells, respectively. Many OX6-immunopositive cells, dendritic or irregular in shape, were recognizable in the lamina propria of oral mucosae: some cells extended their dendritic processes into the epithelial layer of the buccal and sublingual mucosae. Dendritic cells within the epithelium showed intense ATPase reaction, indicating they could be categorized as Langerhans cells. A small number of ED1-positive cells existed in the lamina propria, but none were present in the epithelial cell layer. Double staining either with OX6 and ED1 or OX6 and ACPase made it possible to divide the immunocompetent cells in the lamina propria into three types: the OX6-positive cells without ED1 or ACPase-reaction, the OX6-negative cells with ED1 and ACPase-reactions, and the OX6-ED1/ACPase-co-expressing cells, each of which possessed characteristic ultrastructural features demonstrated by immunoelectron microscopy. Taking these findings together with previous reports, these three types of cells were regarded as a dendritic-like cell, a macrophage without antigen-presentation ability, and a macrophage with antigen-presentation ability, respectively. There were regional differences in the distribution and density of these immunocompetent cells; they were densely distributed in order of the buccal and sublingual mucosae, the palatal mucosa, and the dorsal surface of tongue. The region-specific distribution and density of the immunocompetent cells might be due to the histological. structure of each oral mucosa, suggesting the presence of different immune-defense systems among each portion of the oral mucosae.

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  • Expression of estrogen receptor alpha (ER alpha) in the rat temporomandibular joint. 国際誌

    Kazuho Yamada, Kayoko Nozawa-Inoue, Yoshiro Kawano, Shoji Kohno, Norio Amizuka, Toshihiko Iwanaga, Takeyasu Maeda

    The anatomical record. Part A, Discoveries in molecular, cellular, and evolutionary biology   274 ( 2 )   934 - 41   2003年10月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Numerous epidemiological studies have pointed out a higher frequency of temporomandibular disorder (TMD) in women than in men, which indicates the involvement of a sex hormone, such as estrogen, in the pathogenesis of TMD. Although estrogen is known to play pivotal roles in osteoarthrosis or rheumatoid arthritis in systemic joints, there have been few reports about the role of estrogen in the temporomandibular joint (TMJ). The effect of estrogen is generally mediated by the estrogen receptors (ERs) ER alpha (the predominant type) and ER beta. In this study we examined the expression of ER alpha protein and mRNA in the TMJ of adult male rats by immunocytochemistry and in situ hybridization histochemistry. Intense ER alpha immunoreactivity was localized in the synovial lining cells, stromal cells in the articular disc, and chondrocytes in the TMJ. These ER alpha-immunopositive synovial lining cells are characteristic of cytoplasmic processes identified with confocal and immunoelectron microscopy, which indicates that they are synovial type B cells. In situ hybridization histochemistry confirmed intense signals for ER alpha in the synovial lining cells and the sublining fibroblasts at mRNA levels. The nuclei of chondrocytes showed an intense immunoreaction for ER alpha in the maturative and hypertrophic layers of the articular cartilage. In addition to the nuclear localization of ER alpha, a weak immunoreaction appeared in the cytoplasm of some ER alpha-positive cells. These findings support the hypothesis that TMJ tissue-at least in the male rat-has the potential to be an estrogen target tissue.

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  • Involvement of brain-derived neurotrophic factor (BDNF) in the development of periodontal Ruffini endings 査読

    N Hoshino, F Harada, BA Alkhamrah, M Aita, Y Kawano, K Hanada, T Maeda

    ANATOMICAL RECORD PART A-DISCOVERIES IN MOLECULAR CELLULAR AND EVOLUTIONARY BIOLOGY   274A ( 1 )   807 - 816   2003年9月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:WILEY-LISS  

    The periodontal Ruffini ending has been reported to show immunoreactivity for tyrosine kinase B (trkB), the high-affinity receptor for brain-derived neurotrophic factor (BDNF), in the periodontal ligament of the rat incisor. Furthermore, adult heterozygous BDNF-mutant mice showed malformation and reduction of the periodontal Ruffini endings. To investigate further roles of BDNF in these structures, the development, distribution, and terminal morphology of Ruffini endings were examined in the incisor periodontal ligament of heterozygous and homozygous BDNF mutant mice, as well as in the wild-type littermate by immunohistochemistry for protein gene product (PGP) 9.5, a general neuronal marker. A similar distribution and terminal formation of PGP 9.5-immunoreactive nerve fibers was recognized in the periodontal ligament of all phenotypes at postnatal week (PW) 1. At this stage, the nerve fibers had a beaded appearance, but did not form the periodontal Ruffini endings. At PW2, the heterozygous and wild-type mice started to show ramified nerve fibers resembling the mature shape of periodontal Ruffini endings. At PW3, the Ruffini endings occurred in the periodontal ligament of the wild-type and heterozygous mice. While the Ruffini endings of the wild-type mice appeared either ruffled or smooth, as reported previously, most of these structures showed a smooth outline in the heterozygous mice. The homozygous mice lacked the typical Ruffini endings at PW3. In the quantitative analysis, homozygous mice had the smallest percentages of PGP 9.5-immunoreactive areas at the same postnatal periods, but there were no significant differences between wild-type and heterozygous mice during PW1-3. These findings suggest a possible involvement of BDNF during the postnatal development and, in particular, the maturation of periodontal Ruffini endings. Furthermore, other neurotrophins may play a role in the development and/or early maturation of the periodontal nerve fibers, as indicated by the presence of nerve fibers in the BDNF-homozygous mice. (C) 2003 Wiley-Liss, Inc.

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  • Temporal expression of immunoreactivity for heat shock protein 25 (Hsp25) in the rat periodontal ligament following transection of the inferior alveolar nerve 査読

    K Iijima, F Harada, K Hanada, K Nozawa-Inoue, M Aita, Y Atsumi, S Wakisaka, T Maeda

    BRAIN RESEARCH   979 ( 1-2 )   146 - 152   2003年7月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:ELSEVIER SCIENCE BV  

    The present study examined the immunohistochemical localization of heat shock protein 25 (Hsp25) during the regeneration of nerve fibers and Schwann cells in the periodontal ligament of the rat lower incisor following transection of the inferior alveolar nerve. In the untreated control group, the periodontal ligament of rat incisor did not contain any Hsp25-immunoreaction. On postoperative day 3 (PO 3d), a small number of Schwann cells with slender cytoplasmic processes exhibited Hsp25-immunoreactivity. From PO 5d to PO 21d, Hsp25-positive nerve fibers and Schwann cells drastically increased in number in the alveolar half of the ligament. Although the axons of some regenerating Ruffini-like endings also showed Hsp25-immunoreactions, the migrated Schwann cells were devoid of Hsp25-immunoreaction. Thereafter, Hsp25-positive structures decreased in number gradually to disappear from the periodontal ligament by PO 56d. This temporal expression of Hsp25 in the periodontal ligament well-reflected the regeneration process of the nerve fibers. Hsp25 in the regenerating nerve fibers and denervated Schwann cells most likely serves in modulating actin dynamics and as a cellular inhibitor of apoptosis, respectively. (C) 2003 Elsevier B.V. All rights reserved.

    DOI: 10.1016/S0006-8993(03)02889-0

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  • Repetitive afferent stimulation propagates excitation in the trigeminal caudalis 査読

    K Seo, N Fujiwara, K Takeuchi, T Maeda, G Someya

    NEUROREPORT   14 ( 10 )   1321 - 1325   2003年7月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:LIPPINCOTT WILLIAMS & WILKINS  

    A high-speed optical imaging technique was employed for visualizing neuronal excitation propagation elicited by afferent stimulation in the mouse trigeminal caudalis (Vc) to clarify the central nociceptive modulation mechanism. Membrane depolarization evoked by a single-pulse stimulation to the spinal trigeminal tract (Tr) was propagated rostrally to the Vc, which was suppressed by CNQX This is consistent with our morphological observation that axons expand from the Tr into the Vc. A trained-pulse (tetanus) stimulation to the Tr evoked a broad, persistent excitation in the Vc, while MK-801 suppressed it. Neonatally capsaicin-treated mice maintained a single-pulse response but a lacked tetanus-evoked one. These indicated that prolonged depolarization elicited by repetitive stimulation is a prerequisite to C-fiber excitation for activating the NMDA receptors.

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  • 【軟骨の細胞生物学】PTHrPと軟骨細胞

    網塚 憲生, 下村 淳子, 関 雪絵, 前田 健康

    腎と骨代謝   16 ( 3 )   217 - 227   2003年7月

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    記述言語:日本語   出版者・発行元:(株)日本メディカルセンター  

    副甲状腺ホルモン関連ペプチド(PTHrP)シグナルが軟骨形成に重要な役割を演ずることが明らかにされ,特にPTHrPとIndian hedgehogのnegative feedbackによって軟骨分化が調節されることが明らかとなっている.又,PTH/PTHrP受容体のpoint mutationでJansen型及びBlomstrand型の軟骨異形成症が発症すること,更に内軟骨腫が発症することも明らかにされている.近年の報告では,PTH/PTHrP受容体にNa+/H+交換体調節因子が結合することにより,PTH/PTHrP受容体からのホスフォリパーゼCのシグナル伝達が活性化することが発見された.これら軟骨細胞の分化制御機構について解説した

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  • The involvement of brain-derived neurotrophic factor (BDNF) in the regeneration of periodontal Ruffini endings following transection of the inferior alveolar nerve 査読

    F Harada, N Hoshino, K Hanada, Y Kawano, Y Atsumi, S Wakisaka, T Maeda

    ARCHIVES OF HISTOLOGY AND CYTOLOGY   66 ( 2 )   183 - 194   2003年5月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:INT SOC HISTOLOGY & CYTOLOGY  

    The present study employed immunohistochemistry for protein gene product 9.5 (PGP 9.5) to examine the regeneration process of Ruffini endings, the primary mechanoreceptor in the periodontal ligament, in heterozygous mice with targeted disruption of the brain-derived neurotrophic factor (BDNF) gene and their littermates, following transection of the inferior alveolar nerve. When immunostained for PGP 9.5, periodontal Ruffini endings appeared densely distributed in the periodontal ligament of the heterozygous mice, but the density of the positively stained nerve fibers in the ligament was 20% lower than that in the control littermates. At 3 days after surgery, the PGP 9.5-positive neural elements had disappeared; they began to appear in the periodontal ligament of both animals at 7 days. However, the recovery pattern of the PGP 9.5-positive nerves differed between heterozygous and wild type mice, typical periodontal Ruffini endings morphologically identical to those in the control group appeared in the wild-type mice at 7 days, whereas such Ruffini endings were detectable in the heterozygous mice at 28 days, though much smaller in number. On day 28, when PGP 9.5-positive nerves were largely regenerated in wild type mice, their distribution was much less dense in the ligament of the heterozygous mice than in the non-treated heterozygous mice. The density of PGP 9.5-positive nerve fibers was significantly lower in the heterozygous mice than in wild type mice at any stage examined. These data showing that a reduced expression of BDNF causes delayed regeneration of the periodontal Ruffini endings suggest the involvement of BDNF in the regeneration process of these mechanoreceptors.

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  • Microscopy Research and Technique: Introduction 査読

    Takeyasu Maeda

    Microscopy Research and Technique   60 ( 5 )   467 - 468   2003年4月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    DOI: 10.1002/jemt.10286

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  • Regeneration of nerve fibres in the peri-implant epithelium incident to implantation in the rat maxilla as demonstrated by immunocytochemistry for protein gene product 9.5 (PGP9.5) and calcitonin gene-related peptide (CGRP) 査読

    N Fujii, H Ohnishi, M Shirakura, S Nomura, H Ohshima, T Maeda

    CLINICAL ORAL IMPLANTS RESEARCH   14 ( 2 )   240 - 247   2003年4月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:BLACKWELL MUNKSGAARD  

    The response of nerve fibres in the peri-implant epithelium to titanium implantation was investigated with an experimental model using rat maxilla and immunohistochemical techniques. The latter employed antibodies to protein gene product 9.5 (PGP9.5), and to calcitonin gene-related peptide (CGRP). In control rats without an implantation, a dense innervation of PGP9.5- and CGRP-positive nerve fibres was recognized throughout the junctional epithelium, as has been previously reported. A titanium-implantation induced a remarkable inflammatory reaction, as well as the destruction of covering epithelial cells. By 3-5 days post-implantation, inflammatory reaction showed a tendency to disappear, and the peri-implant epithelium showed proliferation and down-growth along the implant. At this stage, no nerve fibres were found around the peri-implant epithelium. At 10 days, a few nerve fibres reached the basal cell layers of the peri-implant epithelium, and entered it 15 days after implantation when the peri-implant epithelial cells showed morphological features roughly resembling those of normal junctional epithelial cells. At the complete osseointegration stage (days 20-30), the PGP9.5- and CGRP-positive nerve fibres, thin and beaded in appearance, were found distributed in the peri-implant epithelium. After 20 days, the numerical density of the intraepithelial nerves in the peri-implant epithelium appeared the same as, or less than, that in the normal junctional epithelium. These findings indicate that the peri-implant epithelium shows the same innervation as that in normal junctional epithelium, and that the intraepithelial nerve fibres in the peri-implant epithelium might have diverse functions, which have been suggested in the literature.

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  • Dental neuroplasticity, neuro-pulpal interactions, and nerve regeneration 査読

    MR Byers, H Suzuki, T Maeda

    MICROSCOPY RESEARCH AND TECHNIQUE   60 ( 5 )   503 - 515   2003年4月

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    記述言語:英語   出版者・発行元:WILEY-LISS  

    This review covers current information about the ability of dental nerves to regenerate and the role of tooth pulp in recruitment of regenerating nerve fibers. In addition, the participation of dental nerves in pulpal injury responses and healing is discussed, especially concerning pulp regeneration and reinnervation after tooth replantation. The complex innervation of teeth is highly asymmetric and guided towards specific microenvironments along blood vessels or in the crown pulp and dentin. Pulpal products such as nerve growth factor are distributed in the same asymmetric gradients as the dentinal sensory innervation, suggesting regulation and recruitment of those nerve fibers by those specific factors. The nerve fibers have important effects on pulpal blood flow and inflammation, while their sprouting and cytochemical changes after tooth injury are in response to altered pulpal cytochemistry. Thus, their pattern and neuropeptide intensity are indicators of pulp status, while their local actions continually affect that status. When denervated teeth are injured, either by pulp exposure on the occlusal surface or by replantation, they have more pulpal necrosis than occurs for innervated teeth. However, small pulp exposures on the side of denervated crowns or larger lesions in germ-free animals can heal well, showing the value of postoperative protection from occlusal trauma or from infection. Current ideas about dental neuroplasticity, neuro-pulpal interactions, and nerve regeneration are related to the overall topics of tooth biomimetics and pulp/dentin regeneration.

    DOI: 10.1002/jemt.10291

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  • [Mineralization and vascular invasion during endochondral bone formation]. 査読

    Amizuka N, Shimomura J, Maeda T, Ozawa H

    Clinical calcium   13 ( 4 )   405 - 412   2003年4月

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    記述言語:日本語   掲載種別:研究論文(学術雑誌)  

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  • Pulpal regeneration after cavity preparation, with special reference to close spatio-relationships between odontoblasts and immunocompetent cells 査読

    H Ohshima, K Nakakura-Ohshima, K Takeuchi, M Hoshino, Y Takano, T Maeda

    MICROSCOPY RESEARCH AND TECHNIQUE   60 ( 5 )   483 - 490   2003年4月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:WILEY-LISS  

    The regeneration process of the odontoblast cell layer incident to tooth injury, especially its relationship with immunocompetent cells in pulp healing, has not been fully understood. The purpose of the present study was to clarify this relationship between odontoblasts and immunocompetent cells in the process of pulp regeneration following cavity preparation in rat molars by immunocytochemistry for heat shock protein (Hsp) 25 as well as class II major histocompatibility complex (MHC) molecules. In untreated control teeth, intense Hsp 25-immunoreactivity was found in the cell bodies of odontoblasts and their processes within the predentin, whereas class II MHC-positive cells were predominantly located beneath the odontoblast cell layer. Cavity preparation caused the destruction of the odontoblast layer to form an edematous lesion and the shift of class II MHC-positive cells with the injured odontoblasts toward the pulp core at the affected site. Some damaged odontoblasts without apparent cytoplasmic processes, round in profile, retained the immunoreactivity for Hsp25, suggesting the survival of a part of the odontoblasts against artificial external stimuli. Twelve hours after cavity preparation, numerous class II MHC-positive cells appeared along the pulp-dentin border and extended their processes deep into the exposed dentinal tubules. By postoperative 72 hours, newly differentiated odontoblasts with Hsp 25-immunoreactivity were arranged at the pulp-dentin border, but the class II MHC-positive cells moved from the pulp-dentin border to the subodontoblastic layer. These findings indicate that the time course of changes in the expression of Hsp 25-immunoreactivity reflects the regeneration process of odontoblasts. The functional roles of Hsp 25-positive odontoblasts and immunocompetent cells such as class II MHC-positive cells in the process of pulp regeneration after cavity preparation are discussed in conjunction with our previous experimental data.

    DOI: 10.1002/jemt.10289

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  • オステオプロテジェリン欠損マウスにおける骨芽細胞活性と骨改造現象について

    下村 淳子, 網塚 憲生, 関 雪絵, 野田 忠, 前田 健康

    解剖学雑誌   78 ( Suppl. )   191 - 191   2003年4月

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    記述言語:日本語   出版者・発行元:(一社)日本解剖学会  

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  • メカニカルストレスが軟骨細胞に及ぼす影響について

    網塚 憲生, 関 雪絵, 下村 淳子, 監物 新一, 前田 健康

    解剖学雑誌   78 ( Suppl. )   190 - 190   2003年4月

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    記述言語:日本語   出版者・発行元:(一社)日本解剖学会  

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  • The periodontal Ruffini endings in brain derived neurotrophic factor (BDNF) deficient mice 査読

    BA Alkhamrah, N Hoshino, Y Kawano, F Harada, K Hanada, T Maeda

    ARCHIVES OF HISTOLOGY AND CYTOLOGY   66 ( 1 )   73 - 81   2003年3月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:INT SOC HISTOLOGY & CYTOLOGY  

    Innervation and terminal morphology in the lingual periodontal ligament of the incisor were investigated in brain derived neurotrophic factor (BDNF) heterozygous mice and littermate wild-type mice (aged two months) using immunohistochemistry for protein gene product 9.5 (PGP-9.5), a general neuronal marker. In addition, computerassisted quantitative analysis was performed for a comparison of neuronal density in the periodontal ligament between heterozygous and wild-type mice. In wild-type mice, the periodontal ligament was found to be richly innervated by the mechanoreceptive Ruffini endings and nociceptive free nerve endings in the alveolus-related part of the periodontal ligament. The periodontal Ruffini endings in the wild-type mice incisor ligament were classified into two types: type I with ruffled outlines, and type II with a smooth outline. BDNF heterozygous mice showed malformations of the type I Ruffini endings which included fewer nerve fibers and fewer ramifications than those in wild-type mice as well as smooth outlines of the axon terminals. Quantitative analysis under a confocal microscope showed a roughly 18% reduction in neuronal density in the periodontal ligament of the heterozygous mice. These findings suggest that the development and maturation of the periodontal Ruffini endings require BDNF.

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  • 【骨の細胞と形態機能】軟骨内骨化における石灰化と血管侵入

    網塚 憲生, 下村 淳子, 前田 健康, 小澤 英浩

    Clinical Calcium   13 ( 4 )   405 - 412   2003年3月

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    記述言語:日本語   出版者・発行元:(株)医薬ジャーナル社  

    骨端軟骨は大きく,静止層,増殖層,肥大化層に分けられる.肥大化層のカラム間基質は石灰化を受けるが,カラム内の横隔壁は石灰化をあまり受けない.そのために,石灰化軟骨基質は軟骨の長軸方向に形成される.血管内皮細胞は軟骨・骨境界部であまり石灰化を受けていない横隔壁を貫通することで,軟骨に侵入してゆき,その結果,石灰化したカラム間基質が骨組織へと露出する.この露出した軟骨基質は骨芽細胞が接着する足がかりとして役立つと考えられる.又,これらの骨芽細胞は骨基質を軟骨基質上へ分泌することにより,一次骨梁を形成する

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  • Different distribution of immunocompetent cells in the dentogingival junction during root formation in rat molars 査読

    H Tamura, K Nakakura-Ohshima, T Maeda, H Ohshima

    JOURNAL OF PERIODONTAL RESEARCH   38 ( 1 )   10 - 19   2003年2月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:BLACKWELL MUNKSGAARD  

    The distribution of immunocompetent cells in the dentogingival junction of rat molars during root formation was investigated by immunocytochemistry using antibodies to class II major histocompatibility complex (MHC) molecules (OX6-antibody) and monocyte/macrophage lineage cells (ED1-antibody) as well as by histochemical reaction for periodic acid-Schiff (PAS). Two portions (the functional epithelium in the mesial gingiva of the first molar, and the interdental gingiva between the first and second molars) were selected for observations. At the eruption stage of the first molar (16-18 days after birth), OX6-positive cells, dendritic or oval in shape, were abundantly distributed in the connective tissue between the oral epithelium and tooth germ. Positive cells with slender cell processes were also found beneath the ameloblast layer. At the commencement stage of the first molar occlusion (24-28 days after birth), numerous OX6-positive cells displaying a dendritic fashion existed preferentially in the mesial gingiva, but were fewer in the interdental gingiva. In contrast, the interdental gingiva showed a denser distribution of ED1-positive cells and PAS-reactive polymorphonuclear leukocytes (PMLs) than the mesial gingiva. At the completion stage of root formation (100-120 days after birth), the OX6-immunopositive cells invaded the deeper position of the mesial gingiva with the downgrowth of the epithelium; they had a considerably higher cell density compared with those in the interdental gingiva where PAS-reactive PMLs persisted. These findings indicated that the immunocompetent cells showed a region-specific distribution and cell density by their roles in immune response.

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  • Immunohistochemical localization of periostin in developing long bones of mice 査読

    Y Hirose, H Suzuki, N Amizuka, J Shimomura, Y Kawano, K Nozawa-Inoue, A Kudo, T Maeda

    BIOMEDICAL RESEARCH-TOKYO   24 ( 1 )   31 - 37   2003年2月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:BIOMEDICAL RESEARCH PRESS LTD  

    The immunolocalization of periostin, previously termed osteoblast-specific factor 2, was investigated in developing long bones of 17-day-old fetal mice and of 1-, 2-, 3- and 8-week-old mice at the light and electron microscopic levels. Fetal femurs showed immunoreactions for periostin in the periosteum, perichondrium, articular surface of the epiphyseal cartilage, joint ligaments, and fascias of surrounding muscles. In particular, intense immunoreactivity for periostin was found in the fibrous layer of the periosteum and perichondrium. At postnatal 1-and 2-weeks, in contrast, the immunoreactivity was restricted to the periosteum and thick fascias of surrounding muscles when compared with the fetal bones. Immunoelectron microscopic observation of the periosteum demonstrated immunoreaction products for periostin at the junction of periosteal fibroblasts and collagen bundles, suggesting its competence in the cell-to-matrix interaction. Mice at 3 and 8 weeks, unlike 2-week-old mice, showed a periostin-immunoreaction dominantly in the osteoblastic layer but not in the fibroblastic layer of the periosteum. Furthermore, the perichondrium and fascias of surrounding muscles were devoid of immunoreaction. Thus, periostin was confirmed to be widely distributed in bone and concomitant tissues at the fetal stage. As mice grew, however, its immunoreactivity gradually came to be restricted to the osteoblastic layer of the periosteum. Our findings suggest that periostin acts at the site of the cell-to-matrix interaction in periosteum, fascias, and joint ligament during morphogenesis of these tissues.

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  • 歯根膜ルフィニ神経終末の再生・発生過程 査読

    前田健康, 原田史子

    新潟歯誌   33 ( 2 )   167 - 181   2003年

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  • 問題発見・解決型(Problem-Based Learning: PBL)教育法に関する研究−歯学教育の国際化を目指して− 査読

    前田健康, 千田 彰, 松久保隆, 村上俊樹, 吉山昌弘

    日歯教誌   19 ( 1 )   212 - 219   2003年

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  • Defective bone remodelling in osteoprotegerin-deficient mice 査読

    N Amizuka, J Shimomura, MQ Li, Y Seki, M Oda, JE Henderson, A Mizuno, H Ozawa, T Maeda

    JOURNAL OF ELECTRON MICROSCOPY   52 ( 6 )   503 - 513   2003年

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:OXFORD UNIV PRESS  

    Previous studies have reported enhanced osteoclastogenesis, increased bone resorption and osteoporosis in osteoprotegerin (OPG)-deficient mice. In the present study, we show that the tibial epiphyses contain abundant, thin trabeculae lined with numerous osteoclasts and cuboidal osteoblasts. The increase in osteoblasts and osteoclasts was associated with a dramatic increase in calcein labelling of the mineralization fronts and replacement of much of the intertrabecular marrow with numerous alkaline phosphatase-positive preosteoblasts. Furthermore, the discrete, linear cement lines seen in wild-type mice were replaced by a randomly oriented meshwork of cement lines that were stained intensely for tartrate-resistant acid phosphatase and osteopontin in the OPG(-/-) mice. These indices of accelerated bone remodelling in mutant bone were associated with irregular trabecular surfaces, a disorganized collagen matrix interspersed with amorphous ground substance and numerous fissures between old and new bone. In total, these observations indicate that enhanced osteoclastic activity in OPG(-/-) epiphyses led to a coupled increase in osteoblast differentiation and activity and an increase in bone remodelling. The high bone turnover, disorganized matrix and impaired attachment of new to old bone in the cement lines in OPG(-/-) mice appear to cause bone fragility.

    DOI: 10.1093/jmicro/52.6.503

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  • Biological action of parathyroid hormone (PTH)-related peptide (PTHrP) mediated either by the PTH/PTHrP receptor or the nucleolar translocation in chondrocytes. 査読

    Amizuka N, Oda K, Shimomura J, Maeda T

    Anatomical science international   77 ( 4 )   225 - 236   2002年12月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    DOI: 10.1046/j.0022-7722.2002.00032.x

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  • ラット顎関節におけるエストロゲンレセプターαの免疫組織化学的研究

    山田 一穂, 河野 正司, 野澤 佳世子[井上], 網塚 憲生, 前田 健康

    新潟歯学会雑誌   32 ( 2 )   353 - 353   2002年12月

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    記述言語:日本語   出版者・発行元:新潟歯学会  

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  • マウス骨格組織におけるperiostinの局在について

    広瀬 泰史, 鈴木 啓展, 網塚 憲生, 前田 健康

    新潟歯学会雑誌   32 ( 2 )   355 - 355   2002年12月

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    記述言語:日本語   出版者・発行元:新潟歯学会  

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  • 自己硬化型骨補填材(バイオペックス)に対する骨組織の反応

    羽尾 博嗣, 網塚 憲生, 野村 修一, 前田 健康

    新潟歯学会雑誌   32 ( 2 )   342 - 342   2002年12月

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    記述言語:日本語   出版者・発行元:新潟歯学会  

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  • [Not Available]. 査読

    Amizuka N, Seki Y, Maeda T

    Clinical calcium   12 ( 6 )   835 - 843   2002年6月

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    記述言語:日本語   掲載種別:研究論文(学術雑誌)  

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  • 癌とカルシウム 癌の骨転移の微細構造

    網塚 憲生, 関 雪絵, 前田 健康

    Clinical Calcium   12 ( 6 )   835 - 843   2002年5月

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    記述言語:日本語   出版者・発行元:(株)医薬ジャーナル社  

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  • Expression of galanin receptor-1 (GALR1) in the rat trigeminal ganglia and molar teeth 査読

    H Suzuki, T Iwanaga, H Yoshie, J Li, K Yamabe, N Yanaihara, U Langel, T Maeda

    NEUROSCIENCE RESEARCH   42 ( 3 )   197 - 207   2002年3月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:ELSEVIER SCI IRELAND LTD  

    The expression of galanin receptor-1 (GALR1) was investigated in the rat trigeminal ganglion by using immunocytochemistry and in situ hybridization. In addition, the regional distribution of GALR1-immuno reactive pulpal nerves and their ultrastructure were examined in the molar teeth. In the trigeminal ganglion. the immunoreactivity for GALR1 was recognizable in about 30%. of the total number of neurons. Most of the cell bodies were small to medium in size. Analysis of serially cut sections alternately stained with GALR1 and galanin antisera demonstrated that some GALR1-positive cells displayed immunoreactivity for galanin. In situ hybridization analysis, expression of GALR1 mRNA was detected in trigeminal ganglion cells. The cell size distribution was similar to that of GALR1-immunoreactive cells, In the dental pulp. a small number of nerve fibers displayed immunoreactivity for GALR1. The labeled fibers formed terminal arbors in the coronal pulp around and within the odontoblast cell layer, but never penetrated into the predentin and dentin. Ultrastructurally, GALR1 immunoreactivity in the dental pulp was confined to the axoplasm of unmyelinated nerve fibers. The present study provided new evidence that unmyelinated primary afferents innervating dental pulp possessed galanin receptor. and suggests the existence of nociceptive primary afferents functioning as autocrine cells. (C) 2002 Elsevier Science Ireland Ltd and the Japan Neuroscience Society. All rights reserved.

    DOI: 10.1016/S0168-0102(01)00323-6

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  • [The biological role of blood vessels during endochondral bone formation]. 査読

    Amizuka N, Sasaki T, Maeda T

    Clinical calcium   12 ( 3 )   327 - 336   2002年3月

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    記述言語:日本語   掲載種別:研究論文(学術雑誌)  

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  • 【骨と血管の異常 遺伝子からのアプローチ】軟骨内骨化における血管の役割

    網塚 憲生, 佐々木 朝代, 前田 健康

    Clinical Calcium   12 ( 3 )   327 - 336   2002年2月

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    記述言語:日本語   出版者・発行元:(株)医薬ジャーナル社  

    軟骨内骨化を担う細胞群の主役の1つとして血管内皮細胞があげられるが,血管内皮細胞の軟骨侵入に至る迄の軟骨細胞の整然とした細胞増殖・分化は重要なプロセスである.石灰化軟骨基質は軟骨カラム間に形成されており,血管内皮細胞はカラム中の石灰化されていない薄い横隔壁を貫通することで軟骨に侵入すると考えられる.軟骨内骨化における軟骨細胞と血管内皮細胞に焦点をあてて解説した

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  • 歯学教育の改善のために 査読

    脇田 稔, 前田健康, 北田泰之, 深江 允, 下野正基, 吉村文信, 大谷啓一

    歯基礎誌   44 ( 4 )   293 - 303   2002年

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  • 新潟大学歯学部卒前教育における客観的臨床能力試験(OSCE)の導入. 査読

    興地隆史, 福島正義, 葭原明弘, 子田晃一, 小野和宏, 小林 博, 魚島勝美, 小山純市, 植田耕一郎, 高木律男, 前田健康, 花田晃治

    日歯教誌   18 ( 2 )   300 - 307   2002年

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  • Merkel-like cells in Malassez epithelium in the periodontal ligament of cats: An immunohistochemical, confocal-laser scanning and immuno electron-microscopic investigation 査読

    O. Tadokoro, T. Maeda, K. J. Heyeraas, V. Vandevska-Radunovic, Y. Kozawa, I. Hals Kvinnsland

    Journal of Periodontal Research   37 ( 6 )   456 - 463   2002年

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:Blackwell Munksgaard  

    The cellular heterogeneity of Malassez epithelium (ME) residing in the periodontal ligament has recently been reported, and the presence and coexistence of the neuropeptides calcitonin gene-related peptide (CGRP), substance P (SP) and vasoactive intestinal peptide (VIP) in single cells in ME has been shown (1). However, the identity of these neuroendocrine cells has so far not been verified. This study was undertaken in order to elucidate the identity of the neuroendocrine cells in ME by means of transmission electron microscopy, confocal scanning microscopy and immunohistochemistry using antibodies to protein gene product (PGP) 9.5 and cytokeratin 20 (CK). Gingival tissue was included in the study as a positive control for identification of Merkel-like cells in oral epithelium. CK 20 immunopositive cells were present in both Malassez epithelium and in basal cell layers of gingival epithelium showing a distribution consistent with PGP 9.5 labelled cells in both epithelia. The results from PGP 9.5 immuno electron microscopy clearly evidenced the presence of single, intensely labelled cells and some nerve fibres invested between the Malassez epithelial cells. The conformity of the immunopositive cells in Malassez and gingival epithelium verified by double immunolabelling with PGP 9.5 and CK 20. indicates that the labelled neuroendocrine cells arc identical in ME and in gingival epithelium. This demonstrates that Malassez epithelium not only exhibits neuroendocrine cells, but additionally that the neuroendocrine cells represent Merkel-like cells. © Blackwell Munksgaard, 2002.

    DOI: 10.1034/j.1600-0765.2002.01374.x

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  • Expression of heat-shock protein 25 immunoreactivity in the dental pulp and enamel organ during odontogenesis in the rat molar. 国際誌

    Hayato Ohshima, Kuniko Nakakura-Ohshima, Takeyasu Maeda

    Connective tissue research   43 ( 2-3 )   220 - 3   2002年

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    The present immunocytochemical study reports on the expression of heat-shock protein (Hsp) 25 during odontogenesis in rat molars from postnatal 1 to 100 days. Hsp 25 immunoreactivity (IR) appeared in the immature dental mesenchymal cells and the differentiating and differentiated odontoblasts. At 30 days, the coronal odontoblasts retained intense Hsp25-IR, whereas the odontoblasts in the root and floor pulp were initially weak or negative but increased in IR in the later stages, indicating that the expression of Hsp 25 reflects the differentiation status of odontoblasts. During amelogenesis, the secretory ameloblasts were Hsp 25 immunopositive and the enamel free area (EFA) cells showed intense Hsp 25-IR when they developed a ruffled border. Ruffle-ended ameloblasts (RA) also consistently showed intense Hsp 25-IR, but smooth ended ameloblasts (SA) showed weak IR. These data suggest that Hsp 25 is related to the formation and maintenance of the ruffled border of RA and EFA cells.

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  • Functional significance of Msx2 gene during tooth development 査読

    H Ohshima, T Maeda, Satokata, I, R Maas

    DENTIN/PULP COMPLEX   11 - 14   2002年

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    記述言語:英語   掲載種別:研究論文(国際会議プロシーディングス)   出版者・発行元:QUINTESSENCE PUBLISHING CO INC  

    During tooth development, the expression of Msx2 mRNA is spatiotemporally shifted among the several components of the dental epithelium and mesenchyme. Recently, we demonstrated that Msx2-deficient mice display defective tooth, hair follicle, and mammary gland development, in addition to defects of skull ossification and persistent calvarial foramen. Although Msx2-deficient mice have shown the abnormalities of amelogenesis, the detailed phenotypes remain to be clarified. The present study analyzed the abnormalities of tooth development in Msx2-deficient mice. The phenotypes of defective teeth observed in the Msx2(-/-) mice were as follows. The enamel organ showed condensed features, preventing the vascular invasion there from inducing the TUNEL-negative degeneration of ameloblasts. Secretory ameloblasts represented abnormal features such as irregular-shaped Tomes's processes and numerous vacuoles in their distal cytoplasm. The enamel organ contained no TUNEL-positive cells, although the temporary appearance of TUNEL-positive cells occurred in the wild-type mice. Both crown and root in the Msx2(-/-) mice showed the irregular-shaped morphology, concomitant with the abnormal development of Hertwig's epithelial root sheath. The amelogenesis imperfecta was caused by the degeneration of ameloblasts and by the disappearance of the enamel-free area in the cusp area of molars. Our results provide evidence that Msx2 plays crucial roles in amelogenesis and the normal morphology of the tooth.

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  • The expression of galanin receptor-1 in the rat trigeminal ganglion 査読

    H Suzuki, H Yoshie, T Maeda

    DENTIN/PULP COMPLEX   180 - 182   2002年

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    記述言語:英語   掲載種別:研究論文(国際会議プロシーディングス)   出版者・発行元:QUINTESSENCE PUBLISHING CO INC  

    The present study reports on the expression of galanin receptor-1 (GALR1) in rat trigeminal ganglion and pulpal nerves by immunohistochemistry and by in situ hybridization histochemistry. Furthermore, the trigeminal neurons expressing GALR1 were counted by a computer image analyzer. In the rat trigeminal ganglion, GALR1-immunoreactivity was recognizable in about 30% of neurons categorized as small to medium sized, this finding being confirmed by in situ hybridization analysis. One of the antisera to GALR1 could label a small number of pulpal nerve fibers; they were distributed throughout the coronal pulp to terminate near the odontoblast cell layer but never penetrated into the predentin and dentin. Ultrastructurally, immunoreactive products for GALR1 were confined to the axoplasm of unmyelinated nerve fibers, and some, but not all, nerve endings near the odontoblasts were also positive in immunoreaction. The expression of GALR1 in the trigeminal ganglion and dental pulp suggested that galanin/GALR1 might play important roles in transmission and/or modulation of pain sensation in rat teeth.

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  • Optical imaging of excitation propagation evoked by stimulation to, the trigeminal caudalis 査読

    K Seo, N Fujiwara, K Takeuchi, T Maeda, G Someya

    NEUROREPORT   12 ( 18 )   3985 - 3988   2001年12月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:LIPPINCOTT WILLIAMS & WILKINS  

    The present study reports on a characteristic spatio-temporal propagation of differential fluorescent images in the rat, brain stem slice by a high-speed optical imaging technique. Coronal or sagittal slices were stained with a voltage-sensitive dye RH-414, and the superficial layer of the trigeminal caudalis (Vc) was then electrically stimulated. The marginal layer and substantia gelatinosa showed larger rostrocaudal excitation than coronal, despite a lack of tract stimulation expansion in either direction. A perfusion of 0.5 mu mol/l TTX, not 10 mu mol/l CNQX suppressed these propagations. These findings suggest that the superficial layer of Vc has spatial differences in neuronal excitation propagation, as evidenced by morphological observations that dendrites in the superficial layers extend in the rostrocaudal direction. NeuroReport 12:3985-3988 (C) 2001 Lippincott Williams & Wilkins.

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  • The exact expression of glial fibrillary acidic protein (GFAP) in trigeminal ganglion and dental pulp 査読

    H Ajima, Y Kawano, R Takagi, M Aita, H Gomi, MR Byers, T Maeda

    ARCHIVES OF HISTOLOGY AND CYTOLOGY   64 ( 5 )   503 - 511   2001年12月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:INT SOC HISTOLOGY & CYTOLOGY  

    The expression in various cell types of peripheral tissues of glial fibrillary acidic protein (GFAP), first discovered as an intermediate filament specific for astrocytes, remains controversial owing to numerous reports of a wide distribution for GFAP-immunoreactivity in various cells. The present study employed immunohistochemistry to investigate the precise expression of GFAP in the dental pulp and trigeminal ganglion of adult rats and wild-type mice as well as GFAP-knockout mice. The exhibition of GFAP-immunoreactivity in the trigeminal ganglion was further examined by a reverse transcription polymerase chain reaction (RT-PCR) technique, and in situ hybridization histochemistry using a specific cRNA probe prepared by us. The immunoreaction for GFAP was recognizable in the axons, Schwann cells, and the fibroblasts in the dental pulp of rats and wild-type littermate mice. However, mice with null mutations in the GFAP gene remained immunoreactive for GFAP in all these locations. Intense GFAP-immunoreactivity was found in a small number of satellite cells in the trigeminal ganglion in all animals examined in this study. RT-PCR analysis demonstrated bands for the GFAP gene corresponding to the length expected from the primer design in the samples of trigeminal ganglion and dental pulp. In situ hybridization histochemistry also showed intense signals for GFAP mRNA in some satellite cells of the trigeminal ganglion, but never in the neurons. These data suggest that the GFAP-immunoreactive molecules in the pulpal axons and fibroblasts react non-specifically with the polyclonal antibody and are probably a closely related type of intermediate filament.

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  • Expression of 25 kDa heat shock protein by synovial type B cells of the mouse temporomandibular joint 査読

    E Andoh, Y Kawano, H Ajima, K Nozawa-Inoue, S Kohno, T Maeda

    ARCHIVES OF ORAL BIOLOGY   46 ( 10 )   947 - 954   2001年10月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:PERGAMON-ELSEVIER SCIENCE LTD  

    Earlier studies have demonstrated immunoreactivity for heat shock protein 25 (Hsp25) in type B synovial lining cells of the rat temporomandibular joint, and also the presence of characteristic cytoplasmic processes in these cells, but it is unclear whether or not the type B cells in other animals possess such elaborate cytoplasmic projections and as there is as yet no evidence for the synthesis of this protein by these cells. For these reasons, the expression of Hsp25 was investigated in the synovial membrane of the mouse temporomandibular joint by immunocytochemistry and by in situ hybridization using a specific cRNA probe, Intense immunoreaction for Hsp25 was found in the cytoplasm of certain synovial lining cells that were identified as type B by immunoelectron-microscopy. These Hsp25-positive cells had slender cytoplast-nic processes, either projecting towards or covering the synovial surface. Morphological differences between cytoplasmic processes seemed to depend on the location of the type B cell bodies. In situ hybridization showed intense signals for Hsp25 mRNA in the synovial lining cells, suggesting that the type B cells produce, rather than resorb, Hsp25. These findings indicate that Hsp25 is a useful marker for the identification of the synovial type B cells in the temporomandibular joint. It is further hypothesized that Hsp25 in type B cells is involved in maintaining their specific profile and epithelial-like arrangement, and in protecting against mechanical stress. (C) 2001 Elsevier Science Ltd. All rights reserved.

    DOI: 10.1016/S0003-9969(01)00052-8

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  • Possible role of heat shock protein (Hsp) 25 in the enamel organ during amelogenesis in the rat molar 査読

    Y Otsuka, K Nakakura-Ohshima, T Noda, T Maeda, H Ohshima

    ARCHIVES OF HISTOLOGY AND CYTOLOGY   64 ( 4 )   369 - 378   2001年10月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:INT SOC HISTOLOGY & CYTOLOGY  

    The postnatal expression of heat shock protein (Hsp) 25 during the amelogenesis of rat molars was investigated by immunocytochemistry and confocal microscopy. The localization pattern of Hsp 25-immunoreactivity in the inner enamel epithelium and ameloblast cell layer of the rat molars was almost identical to that in the rat incisors which we have previously reported: an intense Hsp 25-immunoreactivity, which first appeared in the preameloblasts, was recognized in secretory ameloblasts and ruffle-ended ameloblasts with stage-specific immunointensity. Confocal microscopy with Hsp 25-antibody and rhodamine-labeled phalloidin clearly demonstrated the co-localization of Hsp 25 and actin filaments in the ameloblast layer, supporting our hypothesis that this molecule might serve to reinforce the ameloblast layer during enamel formation as well as the formation and maintenance of the ruffled border in ruffle-ended ameloblasts. Interestingly, the enamel free area cells, which essentially lack the ability for enamel formation, showed the Hsp 25-immunoreactivity during 4-11 days when they developed a ruffled border, but decreased in that immunoreactivity after postnatal 15 days following apoptosis. Since Hsp 25 has been shown to be a specific inhibitor of apoptosis, the enamel-free area cells contribute to determine the outline of dentin at the cusped area. These data support our previous hypothesis on the diverse functions of Hsp 25 in amelogenesis.

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  • Alteration in the expression of heat shock protein (Hsp) 25-immunoreactivity in the dental pulp of rat molars following tooth replantation 査読

    H Ohshima, K Nakakura-Ohshima, H Yamamoto, T Maeda

    ARCHIVES OF HISTOLOGY AND CYTOLOGY   64 ( 4 )   425 - 437   2001年10月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:INT SOC HISTOLOGY & CYTOLOGY  

    The regeneration process of dental pulp following tooth replantation in rat molars was investigated by immuno cyto chemistry for heat shock protein (Hsp) 25 and protein gene product 9.5 (PGP 9.5). In control teeth at postnatal 4 weeks, the odontoblasts showed intense Hsp 25-immunoreactivity in the coronal dental pulp, but little or no immunoreactivity in the root and floor pulp. In contrast, the Hsp, 25-negative odontoblasts in the latter areas displayed immunoreactivity for PGP 9.5. Tooth replantation caused loss of Hsp 25- and PGP 9.5-immunoreactions in the dental pulp during postoperative days 1-3. At postoperative day 5, plump cells with clear nucleoli and several fine processes-presumably newly differentiated odontoblasts-at the pulp-dentin border became immunopositive for Hsp 25. These data suggest that the expression of Hsp 25- and PGP 9.5-immunoreactivity reflects the status of differentiation of the odontoblasts. Furthermore, some pulpal nerve fibers as well as the Schwann cells in the dental pulp, ordinarily negative in Hsp 25-immunoreaction, acquired their immunoreactivity by postoperative day 5, but lost it thereafter, suggesting the involvement of Hsp, 25 in the regeneration of pulpal nerve fibers. In the case of bone-like tissue formation in the pulp space, on the other hand, no Hsp 25-immunoreactive odontoblasts were recognized in the pulp-dentin border. Thus, the alignment of Hsp 25-immunopositive odontoblasts along the pulp-dentin border indicates a decisive factor for inducing the reparative dentin formation after tooth replantation.

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  • Temporal and spatial distribution of Fos protein in the lumbar spinal dorsal horn neurons in the rat with chronic constriction injury to the sciatic nerve 査読

    Y Yamazaki, T Maeda, G Someya, S Wakisaka

    BRAIN RESEARCH   914 ( 1-2 )   106 - 114   2001年9月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:ELSEVIER SCIENCE BV  

    The temporal and spatial expression pattern of Fos protein in spinal dorsal horn neurons was examined by immunohistochemistry in rats with chronic constriction injury (M) to the sciatic nerve. In normal animals, a few Fos-immunoreactive (-IR) neurons were detected in the dorsal horn of the lumbar spinal cord, Following induction of M, a very large number of Fos-IR neurons appeared in the spinal dorsal horn, but a significant number of Fos-IR neurons were also observed in the contralateral dorsal horn where primary afferents of the injured sciatic nerve rarely project. Sham-operated animals also had a significant number of Fos-IR neurons in the dorsal horn bilaterally. The number of Fos-IR neurons reached its maximal level I day following placement of the ligatures (PO ld). The ratio of the number of Fos-IR neurons in the ipsilateral dorsal horn to the contralateral dorsal horn, however, had its peak level 3 days following CCI (3.1-fold increase compared to the contralateral dorsal horn). The number of Fos-IR neurons in the dorsal horn gradually decreased, but increased again around PO 15d. On PO 30d, the number of Fos-IR neurons decreased and became comparable to that in normal animals. The present results indicate that the induction of Fos-IR neurons in the dorsal horn caused by CCI is biphasic and reaches its maximal level on PO 3d, near the time of hyperalgesia onset. (C) 2001 Elsevier Science BY. All rights reserved.

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  • Temporal and spatial distribution of Fos protein in the parabrachial nucleus neurons during experimental tooth movement of the rat molar 査読

    K Hiroshima, T Maeda, K Hanada, S Wakisaka

    BRAIN RESEARCH   908 ( 2 )   161 - 173   2001年7月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:ELSEVIER SCIENCE BV  

    The present study was undertaken to reveal spatio-temporal changes in the distribution of Fos-like immunoreactive (-IR) neurons in the parabrachial nucleus (PBN), one of the important relay nuclei for processing autonomic and somatosensory information from the oro-facial regions, following the induction of experimental tooth movement in rat upper molars. The experimental tooth movement was induced by the insertion of elastic rubber between the first and second upper molars. In normal animals, the PBN contained a smaller number of Fos-IR neurons. Following experimental tooth movement, the Fos-IR neurons increased in number significantly on both the ipsilateral and contralateral PBN, reaching a maximum at 4 h (about 10 times that of normal animals), and then decreased gradually. However, a significant number of Fos-IR neurons remained at 24 h post-operation. Remarkable side-by-side differences in the number of Fos-IR neurons were recognized at 1 to 4 h following the experimental tooth movement. Their number returned to normal (basal) levels at 5 days post. All subnuclei of PBN showed similar temporal changes in the number of Fos-IR neurons, this being particularly apparent in lateral PBN. Administrations of morphine (3 and 10 mg/kg, i.p.) drastically reduced the induction of Fos-IR neurons in all subnuclei of both the ipsilateral and contralateral PBN in a dose-dependent manner, and its effect was antagonized by pretreatment with naloxone (2 mg/kg, i.p.). The reduction of Fos-IR neurons by morphine pretreatment suggests that the appearance of Fos-IR neurons in the PBN may be partly due to the noxious stimulation and/or stress arising from tooth movement. The bilateral expression of Fos-IR neurons in the PBN indicates that the experimental tooth movement causes the activation of PBN neurons for the processing of somatosensory as well as autonomic information. The prolonged expression of Fos-IR neurons in all the subnuclei of bilateral PBN reflects clinical features of the transient discomfort and/or abnormal sensations, which many patients often complain about during orthodontic treatment. (C) 2001 Elsevier Science B.V. All rights reserved.

    DOI: 10.1016/S0006-8993(01)02639-7

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  • Immunocytochemical detection of superoxide dismutases (SODs) in the periodontal Ruffini endings of the rat incisor 査読

    H Yamamoto, S Hayashi, K Nakakura-Ohshima, Y Kawano, K Nozawa-Inoue, H Ohshima, T Maeda

    BRAIN RESEARCH   905 ( 1-2 )   232 - 235   2001年6月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:ELSEVIER SCIENCE BV  

    The expression of immunoreactivities for superoxide dismutases (SODs), Mn-SOD and Cu/Zn-SOD. was immunohistochemically investigated in the lingual periodontal ligament and toe pads of adult rats. Immunocytochemistry for SODs revealed that the axon terminals of both the periodontal Ruffini endings and cutaneous Meissner's corpuscles showed mitochondrial Mn-SOD immunoreactivity, but not cytosolic Cu/Zn-SOD immunoreactivity, indicating Mn-SOD is a useful marker for identifying the mechanoreceptors. It is likely that Mn-SOD in the axon terminals of mechanoreceptors exerts protective action against nerve injury and neuronal death under severe conditions, serving to scavenge free radicals from the axon terminals. (C) 2001 Elsevier Science B.V. All rights reserved.

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  • Responses of immunocompetent cells in the dental pulp to replantation during the regeneration process in rat molars 査読

    A Shimizu, K Nakakura-Ohshima, T Noda, T Maeda, H Ohshima

    CELL AND TISSUE RESEARCH   302 ( 2 )   221 - 233   2000年11月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:SPRINGER-VERLAG  

    Responses of immunocompetent cells to tooth replantation during the regeneration process of the dental pulp in rat molars were investigated by immunocytochemistry using antibodies to class II major histocompatibility complex (MHC) molecules (OX6 antibody), monocyte/macrophage lineage cells (ED1 antibody) and protein gene product 9.5 (PGP 9.5), as well as by histochemical reaction for periodic acid-Schiff (PAS). Tooth replantation caused an increase in both the number of OX6- and ED1-positive cells and their immunointensity in the replanted pulp, but almost all PGP 9.5-immunoreactive nerves diminished in the initial stages. By postoperative day 3, many OX6- and ED1-immunopositive cells had accumulated along the pulp-dentin border to extend their cytoplasmic processes into the dentinal tubules in successful cases. Once reparative dentin formation had begun after postoperative day 7, OX6- and ED1-immmunopositive cells became scattered in the odontoblast layer, while reinnervation was found in the coronal pulp. The temporal appearance of these immunocompetent cells at the pulp-dentin border suggests their participation in odontoblast differentiation as well as in initial defense reactions during the pulpal regeneration process. On postoperative day 14, the replanted pulp showed three regeneration patterns: (1) reparative dentin, (2) bone-like tissue formation, and (3) an intermediate form between these. In all cases, PAS-reactive cells such as polymorphonuclear leukocytes (PML) and mesenchymal cells occurred in the pulp space. However, the prolonged stagnation of inflammatory cells was also discernible in the latter two cases. Thus, the findings on PAS reaction suggest that the migration of the dental follicle-derived cells into the pulp space and the subsequent total death of the proper pulpal cells are decisive factors for eliciting bone-like tissue formation in the replanted pulp.

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  • Transient expression of heat shock protein (Hsp) 25 in the dental pulp and enamel organ during odontogenesis in the rat incisor 査読

    H Ohshima, H Ajima, Y Kawano, K Nozawa-Inoue, S Wakisaka, T Maeda

    ARCHIVES OF HISTOLOGY AND CYTOLOGY   63 ( 4 )   381 - 395   2000年10月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:INT SOC HISTOLOGY & CYTOLOGY  

    The expression of heat shock protein (Hsp) 25 during odontogenesis in the dental pulp and enamel organ of rat incisors was investigated by immunocytochemistry and confocal microscopy. In the process of dentin formation, immature odontoblasts first exhibited Hsp 25-immunoreactivity, and increased in immunointensity with the advance of their differentiation. In the dental pulp, in contrast, intense immunoreaction in the mesenchymal cells became weak or negative in parallel with the progress of cell differentiation. The immunoreaction for Hsp 25 in the enamel organ revealed a characteristic stage-related alteration during amelogenesis. In secretory ameloblasts, the immunoreaction for Hsp 25 was found throughout their cell bodies, intense reactivity being located near the proximal and distal terminal webs. At the maturation stage, ruffle-ended ameloblasts (RA) consistently showed Hsp 25-immunoreactivity throughout the cell bodies, whereas smooth-ended ameloblasts (SA) lacking a ruffled border were weak in immunoreaction at the distal cytoplasm. Other cellular elements of the enamel organ were negative. The subcellular localization of Hsp 25-immunoreactivity in this study appeared essentially identical to that of actin filaments as demonstrated by confocal microscopy using rhodamine-labeled phalloidin. These immunocytochemical data suggest that the Hsp 25 molecule is involved in reinforcement of the cell layer following cell movement during odontogenesis and in the formation and maintenance of the ruffled border of RA.

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  • Msx2 deficiency in mice causes pleiotropic defects in bone growth and ectodermal organ formation

    Satokata, I, L Ma, H Ohshima, M Bei, Woo, I, K Nishizawa, T Maeda, Y Takano, M Uchiyama, S Heaney, H Peters, ZQ Tang, R Maxson, R Maas

    NATURE GENETICS   24 ( 4 )   391 - 395   2000年4月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:NATURE AMERICA INC  

    The composite structure of the mammalian skull, which forms predominantly via intramembranous ossification, requires precise pre- and post-natal growth regulation of individual calvarial elements. Disturbances of this process frequently cause severe clinical manifestations in humans. Enhanced DNA binding by a mutant MSX2 homeodomain results in a gain of function and produces craniosynostosis in humans(1,2). Here we show that Msx2-deficient mice have defects of skull ossification and persistent calvarial foramen, This phenotype results from defective proliferation of osteoprogenitors at the osteogenic front during calvarial morphogenesis, and closely resembles that associated with human MSX2 haploinsufficiency in parietal foramina(3) (PFM), Msx2(-/-) mice also have defects in endochondral bone formation. In the axial and appendicular skeleton, post-natal deficits in Pth/Pthrp receptor (Pthr) signalling and in expression of marker genes for bone differentiation indicate that Msx2 is required for both chondrogenesis and osteogenesis, Consistent with phenotypes associated with PFM, Msx2-mutant mice also display defective tooth, hair follicle and mammary gland development, and seizures, the latter accompanied by abnormal development of the cerebellum. Most Msx2-mutant phenotypes, including calvarial defects, are enhanced by genetic combination with Msx1 loss of function, indicating that Msx gene dosage can modify expression of the PFM phenotype. Our results provide a developmental basis for PFM and demonstrate that Msx2 is essential at multiple sites during organogenesis.

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  • A histochemical investigation of the bone formation process by guided bone regeneration in rat jaws. Effect of PTFE membrane application periods on newly formed bone 査読

    H Ohnishi, N Fujii, T Futami, N Taguchi, H Kusakari, T Maeda

    JOURNAL OF PERIODONTOLOGY   71 ( 3 )   341 - 352   2000年3月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:AMER ACAD PERIODONTOLOGY  

    Background: Guided bone regeneration (GBR) has been widely utilized for the promotion of bone augmentation in bone loss areas. However, little information has been available regarding chronological changes in newly formed bone and alterations in the nature of newly formed bone after removal of a barrier membrane. The present study attempted to establish a GBR model for rat maxillae. We also examined the effects of membrane application periods on newly formed bone and its remodeling process after removal of the membrane in this experimental model.
    Methods: Thirty-five Wistar rats were divided into 2 groups: a membrane application group and a membrane removal group. The chronological changes of newly formed bone were evaluated histologically and statistically.
    Results: At 2 weeks after the GBR procedure, bony cavities had completely filled the newly formed bone in the experimental side. In the control side, corticalization on the surface of the newly formed bone proceeded with a decrease in the bone marrow cavity, whereas the bone marrow space had enlarged by 12 weeks post-surgery in the experimental side. In the membrane removal group, the osteoblasts appeared on newly formed bone at 1 week after membrane removal. Comparatively thick compact bone had formed on the surface of the newly formed bone at 4 weeks after membrane removal, and corticalization proceeded later.
    Conclusions: The long-term application of a barrier membrane induces the enlargement of the bone marrow spaces. We suggest that PTFE membrane removal in adequate time promotes the corticalization and maturation of the newly formed bone by the GBR technique.

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  • The development of terminal Schwann cells associated with periodontal Ruffini endings in the rat incisor ligament 査読

    S Hayashi, K Nakakura-Ohshima, H Ohshima, T Noda, S Honma, S Wakisaka, T Maeda

    BRAIN RESEARCH   858 ( 1 )   167 - 171   2000年3月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:ELSEVIER SCIENCE BV  

    The postnatal development of the terminal Schwann cell, an analogue of the lamellar cell in cutaneous sensory receptors, was examined by histochemistry for non-specific cholinesterase and immunohistochemistry for S-100 protein in the periodontal Ruffini endings of the rat incisor. Double immunohistochemistry for S-100 protein and protein gene product 9.5 (PGP 9.5) was also performed to examine the relationship between terminal Schwann cells and axons. Histochemistry for non-specific cholinesterase was able to demonstrate the age-related development of the terminal Schwann cells. the morphology and distribution of the developing terminal Schwann cells became almost identical to those in adults during postnatal days 15-18. Axons showing PGP 9.5-like immunoreactivity elongated and expanded after arrangement of terminal Schwann cells in the alveolus-related part. This suggests that the terminal Schwann cell is important in the development and maturation of the periodontal Ruffini endings. (C) 2000 Elsevier Science B.V. All rights reserved.

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  • Tissue response to titanium implants in the rat maxilla: Ultrastructural and histochemical observations of the bone-titanium interface 査読

    T Futami, N Fujii, H Ohnishi, N Taguchi, H Kusakari, H Ohshima, T Maeda

    JOURNAL OF PERIODONTOLOGY   71 ( 2 )   287 - 298   2000年2月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:AMER ACAD PERIODONTOLOGY  

    Background: The detailed mechanism of osseointegration, the most appropriate implant-bone interface, remains unclear in jaw tissues at the ultrastructural level in contrast to the many reports using long bones. The present study reports on tissue response to titanium-implantation on an animal model using rat maxilla.
    Methods: Animals were sacrificed at 1 to 28 days postimplantation and prepared tissue specimens, freed from implants by a cryofracture technique, were processed for transmission electron microscopy and histochemistry for tartrate resistant acid phosphatase activity (TRAPase).
    Results: Different patterns in bone formation were recognized between lateral and base areas of implant cavities. In the lateral area with narrow gaps, bone deposition took place from the pre-existing bone towards the implant after active bone resorption by osteoclasts reactive to TRAPase. However, no distinct bone formation appeared in the lateral area where the implant had been installed close to the osteotomy margin. On the other hand, new bone formation was found at the base area without any apparent bone resorption. Interestingly, mononuclear cells reactive to TRAPase, presumably preosteoblasts, frequently occurred near preosteoblasts. Osseointegration around the implants was obtained in this model by 28 days post-implantation except for the lateral area with complete contact with implants, where the thin layer remained in contact with the implant surface.
    Conclusions: These findings indicate that ossification proceeds at different modes around the titanium implant in rat maxilla, depending on the nature of the recipient bones and the dimension of the gap between the implant and osteotomy margin.

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  • Morphological basis of periodontal nerve endings 査読

    Takeyasu Maeda, Sachiko Hayashi

    Acta Anatomica Nipponica   75 ( 4 )   379 - 380   2000年

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    記述言語:日本語   掲載種別:研究論文(学術雑誌)   出版者・発行元:Japanese Association of Anatomists  

    The periodontal ligament, a dense collagenous tissue between tooth and alveolar bone, receives a rich sensory nerve supply, and contains tsvo kinds of sensory receptors
    nociceptor and mechanoreceptor. The mechanical stimuli for teeth can evoke various oral reflexes, which facilitate mastication via the periodontal mechanorecepors. In spite of many reports on the periodontal sensory receptors, recent studies have revealed that the Ruffini endings, categorized as low-threshold slowly adapting type II (SA II) stretch receptors, are primary mechanoreceptors in the periodontal ligament. This paper summarized recent findings on the morphological features and developmental aspects of the periodontal Ruffini endings.

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  • Delayed expression of calbindin D28k during regeneration of the periodontal Ruffini endings of the rat incisor following injury to the inferior alveolar nerve 査読

    Yukako Atsumi, Hisao Ajima, Takeyasu Maeda, Kojiro Kurisu, Satoshi Wakisaka

    Cells Tissues Organs   167 ( 4 )   285 - 289   2000年

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:S. Karger AG  

    Expression of calbindin D28k (CB)-like immunoreactivity (-LI) was compared with that of protein gene product 9.5 (PGP 9.5), a general neuronal marker, in the periodontal ligament of the rat lower incisor following resection of the inferior alveolar nerve (IAN). In normal animals, the periodontal nerve fibers showing PGP 9.5-LI formed either Ruffini endings with expanded arborization or thin free nerve endings in the alveolar half of the ligament. Thick CB-like immunoreactive (-IR) nerve fibers terminated in a dendritic fashion in the same region, but thin CB-IR nerve fibers were rarely detected. During the 3 days following resection of the IAN, most of the PGP 9.5-IR and all CB-IR nerve fibers disappeared. Regenerated PGP 9.5-IR nerve fibers appeared around 7 days after resection, in contrast to the very small number of regenerated CB-IR nerve fibers. Around 21-28 days following resection, the number and terminal morphology of regenerated PGP 9.5-IR nerve fibers were comparable to those observed in normal animals, but the number of regenerated CB-IR nerve fibers was still smaller. The terminal morphologies of these regenerated CB-IR nerve fibers showed less expansion compared with normal animals at these post-injured periods. The number of regenerated CB-IR nerve fibers increased gradually to return to normal by 56 days following injury. The delayed expression of CB in the regenerated periodontal Ruffini endings suggests that the functional recovery of periodontal Ruffini endings occurred after the regeneration of periodontal Ruffini endings had been completed. Copyright (C) 2000 S. Karger AG, Basel.

    DOI: 10.1159/000016791

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  • Effects of different types of injury to the inferior alveolar nerve on the behavior of Schwann cells during the regeneration of periodontal nerve fibers of rat incisor 査読

    Yukako Atsumi, Takumi Imai, Ken Matsumoto, Masayoshi Sakuda, Takeyasu Maeda, Kojiro Kurisu, Satoshi Wakisaka

    Archives of Histology and Cytology   63 ( 1 )   43 - 54   2000年

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:Japan Society of Histological Documentation  

    The present study reports on different regeneration patterns of axons and Schwann cells in the periodontal ligament of the rat incisor using immunohistochemistry of protein gene product 9.5 (PGP 9.5) and S-100 protein. Three kinds of injury (transection, crush and segmental resection) were applied to the inferior alveolar nerve. In normal animals, PGP 9.5- and S- 100-immunoreactivities were detected in the axons and Schwann cell elements of periodontal Ruffini endings, respectively. They were restricted to the alveolus-related part, occurring only rarely in the tooth-related part and in the shear zone (the border between the alveolus-related and tooth-related parts). Both transection and segmental resection caused the complete disappearance of PGP 9.5-immunoreactive nerve fibers in the periodontal ligament, while a small number of them could be found following the crush injury. Regenerating PGP 9.5-reactive nerve fibers appeared at 5 days and 21 days following the transection and segmental resection, respectively. The regeneration of periodontal nerve fibers completed in a period of 21-28 days and 14-21 days following the transection and crush, respectively, but was not completed even at 56 days following the segmental resection. The behavior of Schwann cells during regeneration was similar after the different nerve injuries
    spindle-shaped S-100-immunoreactive cells, presumably Schwann cells, appeared in the shear zone and the tooth-related part. These cells disappeared 5-7 days prior to the complection of the regeneration of axonal elements of the periodontal ligament following the transection and crush. Following the segmental resection, in contrast, spindle-shaped S-100-positive cells disappeared from the tooth-related part at 42 days, although the axonal regeneration of periodontal Ruffini endings proceeded even until 56 days. We thus conclude that the duration of the migration of Schwann cells depends on the state of the regeneration of axons.

    DOI: 10.1679/aohc.63.43

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  • Altered distribution of Schwann cells in the periodontal ligament of the rat incisor following resection of the inferior alveolar nerve: An immunohistochemical study on S-100 proteins 査読

    Yukako Atsumi, Ken Matsumoto, Masayoshi Sakuda, Takeyasu Maeda, Kojiro Kurisu, Satoshi Wakisaka

    Brain Research   849 ( 1-2 )   187 - 195   1999年12月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    The present study employed immunohistochemistry for the detection of S-100 proteins to reveal the alteration in the distribution of Schwann cells in the periodontal ligament of the rat incisor following resection of the inferior alveolar nerve (IAN). In normal animals, S-100-immunostaining demonstrated the profiles of Ruffini endings, primary mechanoreceptors in the periodontal ligament, in the alveolus-related part of the ligament. Under the electron microscope, S-100-like immunoreactivity (-LI) was observed in the cytoplasm of the terminal Schwann cell elements and in some axon profiles of the Ruffini endings. During the regeneration, S-100-like immunoreactive (-IR) terminal Schwann cells in the alveolus-related part of the ligament gradually decreased in number. In contrast, S-100-LI was found in the spindle-shaped cells at the shear zone (the border between alveolus-related and tooth-related parts) and in the tooth-related part, where S-100-LI was rarely detected in normal animals. Immunoelectron microscopic observations revealed that some S-100-IR spindle-shaped cells contained fibrous long spacing (FLS) fibers, suggesting that they were Schwann cells. Some regenerating axons were observed at the shear zone, but were rarely found in the tooth-related part. With the progress of the regeneration of the periodontal Ruffini endings, S-100-IR terminal Schwann cells became rearranged in the alveolus-related part by 42-56 days post injury, whereas the S-100-IR spindle-shaped Schwann cells in the shear zone and tooth-related part disappeared when the regeneration was complete. Copyright (C) 1999 Elsevier Science B.V.

    DOI: 10.1016/S0006-8993(99)02153-8

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  • Immunocytochemical demonstration of heat shock protein 25 in the rat temporomandibular joint 査読

    K Nozawa-Inoue, H Ohshima, Y Kawano, H Yamamoto, R Takagi, T Maeda

    ARCHIVES OF HISTOLOGY AND CYTOLOGY   62 ( 5 )   483 - 491   1999年12月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:INT SOC HISTOLOGY & CYTOLOGY  

    The expression of heat shock protein 25 (Hsp 25) was investigated in the rat temporomandibular joint by immunocytochemistry combined with confocal and electron microscopy. Immunostaining with an antibody to Hsp25 was able to demonstrate various cellular elements in the synovial membrane of the joint. Intense immunoreaction for Hsp25 was recognized in certain cells comprising the synovial lining layer. Confocal microscopic observation revealed two characteristic profiles of the Hsp25-positive cells with cytoplasmic processes: one extended thick and long processes towards the articular cavity, and the other prejected horizontally slender processes which covered the synovial membrane. Under the electron microscope, the immunoreactive synovial lining cells were characterized by a well-developed rough endoplasmic reticulum and secretory granules, suggesting that they can be categorized as fibroblastic type B cells. The covering by the cytoplasmic extensions was confirmed by immune-electron microscopic observations. This cytoplasmic covering presumably performs a barrier function and expedites the effective secretion/resorption of synovial fluids. Since it has been proposed that Hsp 25 is associated with an estrogen receptor, the immunopositive synovial lining cells were considered estrogen-target cells. Immunoreactivity for Hsp25 was also observed in the chondrocytes of the maturative and hypertrophic cell layers as well as in the cells of the articular disk. A suggestion was made that Hsp 25 might be involved in the inhibition of apoptosis of those cells.

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  • Heterogeneous localizations of Trk B among individual periodontal Ruffini endings in the rat incisor 査読

    Y Atsumi, S Hayashi, K Nakakura-Ohshima, T Maeda, K Kurisu, S Wakisaka

    ARCHIVES OF HISTOLOGY AND CYTOLOGY   62 ( 5 )   435 - 440   1999年12月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:INT SOC HISTOLOGY & CYTOLOGY  

    The present immunocytochemical study examined the localization of Trk B, a high affinity neurotrophin receptor, in the neural elements of the periodontal ligament of the rat incisor. In light microscopy, the immunoreactivity was demonstrated in dendritic profiles in the alveolar half of the periodontal ligament. Their location and morphological features indicated that they were periodontal Ruffini endings. Occasional rounded cells associated with periodontal Ruffini endings, which had immunonegative kidney-shaped nuclei, were immunoreactive; these were judged to be terminal Schwann cells. Immunoelectron microscopy revealed the heterogeneous localization of Trk B among individual Ruffini endings. Some terminal Schwann cells contained immunoreactive products for Trk B in the cytoplasm, while others did not. Similarly, a part of the Schwann sheaths covering the axon terminals showed Trk B immunoreactivity. Most axon terminals associated with periodontal Ruffini endings were immunopositive for Trk B, though a few of them were immunonegative. The ordinary Schwann cells did not contain Trk B immunoreactive products. These findings imply that Trk B is required for the maintenance of periodontal Ruffini endings, The different expression pattern of Trk B suggests that neuronal and glial elements comprising individual periodontal Ruffini endings are subject to heterogeneous conditions with regard to the requirement of Trk B.

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  • Re-innervation in the canine periodontal ligament of replanted teeth using an antibody to protein gene product 9.5: an immunohistochemical study 査読

    H Yamada, T Maeda, K Hanada, Y Takano

    ENDODONTICS & DENTAL TRAUMATOLOGY   15 ( 5 )   221 - 234   1999年10月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:MUNKSGAARD INT PUBL LTD  

    canine teeth was examined by immunohistochemistry for protein gene product 9.5 (PGP 9.5), a general marker for neurons, and by electron microscopy. Within 1 week of replantation, the periodontal fibers had regenerated, filling the narrow spaces between the alveolar bone and the root surface around the cervical and apical regions. Near the root apex, however, no PGP 9.5-immunopositive nerve fibers were found in the regenerated periodontal ligament except for those in the alveolar half of the ligament. At 2 weeks after replantation, many nerve fibers positive for PGP 9.5 had ascended the periodontal ligament from the thick nerve bundles located near the root apex. Fine nerve endings showing complicated ramification were also present in the apical region. By 3 or 4 weeks after replantation, the vascular network was regenerated and principal periodontal fibers were re-established throughout the entire length of the periodontal ligament. The extensively ramified PGP 9.5-immunopositive structures had increased in thickness and density and showed characteristic treelike profiles by 3 weeks. Electron microscopy confirmed that most of these structures were Ruffini-like endings, and demonstrated that such nerve terminals were almost regenerated by 4 weeks post-replantation. These results indicated that, in the periodontal ligament of replanted canine teeth, the regeneration of the nerve fibers including mechanosensory receptors first showed signs of regeneration by 2 weeks following tooth replantation and proceeded rapidly thereafter. Regeneration of the periodontal ligament including fiber architecture as well as vascular and neural elements was almost complete by 4 weeks after replantation.

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  • The Ruffini ending as the primary mechanoreceptor in the periodontal ligament: Its morphology, cytochemical features, regeneration, and development 査読

    T Maeda, K Ochi, K Nakakura-Ohshima, SH Youn, S Wakisaka

    CRITICAL REVIEWS IN ORAL BIOLOGY & MEDICINE   10 ( 3 )   307 - 327   1999年8月

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    記述言語:英語   出版者・発行元:INT AMER ASSOC DENTAL RESEARCH  

    The periodontal ligament receives a rich sensory nerve supply and contains many nociceptors and mechanoreceptors. Although its various kinds of mechanoreceptors have been reported in the past, only recently have studies revealed that the Ruffini endings-categorized as low-threshold, slowly adapting, type II mechanoreceptors-are the primary mechanoreceptors in the periodontal ligament. The periodontal Ruffini endings display dendritic ramifications with expanded terminal buttons and. Furthermore, are ultrastructurally characterized by expanded axon terminals filled with many mitochondria and by an association with terminal or lamellar Schwann cells. The axon terminals of the periodontal Ruffini endings have finger-like projections called axonal spines or microspikes, which extend into the surrounding tissue to detect the deformation of collagen fibers. The functional basis of the periodontal Ruffini endings has been analyzed by histochemical techniques. Histochemically, the axon terminals are reactive for cytochrome oxidase activity, and the terminal Schwann cells have both non-specific cholinesterase and acid phosphatase activity. On the other hand, many investigations have suggested that the Ruffini endings have a high potential for neuroplasticity. For example, immunoreactivity for p75-NGFR (low-affinity nerve growth factor receptor) and GAP-43 (growth-associated protein-43), both of which play important roles in nerve regeneration/development processes, have been reported in the periodontal Ruffini endings, even in adult animals (though these proteins are usually repressed or down-regulated in mature neurons). Furthermore. in experimental studies on nerve injury to the inferior alveolar nerve, the degeneration of Ruffini endings takes place immediately after nerve injury, with regeneration beginning from 3 to 5 days later, and the distribution and terminal morphology returning to almost normal at around 14 days. During regeneration. some regenerating Ruffini endings expressed neuropeptide Y, which is rarely observed in normal animals. On the other hand, the periodontal Ruffini endings show stage-specific configurations which are closely related to tooth eruption and the addition of occlusal forces to the tooth during postnatal development, suggesting that mechanical stimuli due to tooth eruption and occlusion are a prerequisite for the differentiation and maturation of the periodontal Ruffini endings. Further investigations are needed to clarify the involvement of growth factors in the molecular mechanisms of the development and regeneration processes of the Ruffini endings.

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  • Polypoid mass of the gingiva 査読

    K Izumi, T Nakajima, T Maeda, T Irie, M Murata, T Saku

    ORAL SURGERY ORAL MEDICINE ORAL PATHOLOGY ORAL RADIOLOGY AND ENDODONTICS   88 ( 2 )   117 - 121   1999年8月

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    記述言語:英語   出版者・発行元:MOSBY-YEAR BOOK INC  

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  • Immunocytochemical demonstration of laminin in the synovial lining layer of the rat temporomandibular joint 査読

    K Nozawa-Inoue, H Ajima, R Takagi, T Maeda

    ARCHIVES OF ORAL BIOLOGY   44 ( 6 )   531 - 534   1999年6月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:PERGAMON-ELSEVIER SCIENCE LTD  

    This immunocytochemical study describes the distribution of laminin in the synovial lining of the rat temporomandibular joint. Laminin immunostaining was present around some synovial lining cells and blood vessels. Ultrastructurally, immunoreactive products for laminin were deposited around cells with a well-developed rough endoplasmic reticulum and secretory granules, suggesting that they were type B synovial lining cells. The localization of laminin immunoreactivity was not uniform around the cell membrane, the most intense immunoreaction being present on the basal aspect membrane as is seen in the basement membrane of epithelia. In contrast, macrophage-like synovial lining type A cells did not show laminin immunoreactivity. This different immunostaining pattern suggests that laminin acts as an adhesion molecule for the type B cells in their epitheliallike arrangement. (C) 1999 Elsevier Science Ltd. All rights reserved.

    DOI: 10.1016/S0003-9969(99)00021-7

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  • Postnatal expression of calretinin-immunoreactivity in periodontal ruffini endings in the rat incisor: A comparison with protein gene product 9.5 (PGP-9.5)-immunoreactivity 査読

    T Asahito, H Ohshima, K Hanada, S Wakisaka, T Maeda

    ARCHIVES OF HISTOLOGY AND CYTOLOGY   62 ( 1 )   57 - 69   1999年3月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:INT SOC HISTOLOGY & CYTOLOGY  

    The postnatal expression of immunoreactivity for calretinin, one of the calcium binding proteins, and for protein gene product 9.5 (PGP 9.5), a general neuronal marker, was investigated in mechanoreceptive Ruffini endings in the periodontal ligament of the rat incisor. Age-related changes in the expression of these two proteins in periodontal nerves were further quantified with a computerized image analysis. At 1 day after birth, a few PGP 9.5-immunoreactive nerve fibers and a still smaller number of calretinin-positive fibers were found in the periodontal ligament: they were thin and beaded in appearance and no specialized nerve terminals were recognized, Tree-like terminals, reminiscent of immature Ruffini endings, were recognizable in 4-day-old rats by PGP 9.5-immunohistochemistry, while calretinin-immunostaining failed to reveal these specialized endings. At postnatal 7-11 days when PGP 9.5-immunostaining could demonstrate typical Ruffini endings, calretinin-immunopositive nerve fibers merely tapered off without forming the Ruffni type endings, A small number of Ruffini endings showing calretinin-immunoreactivity began to occur in the periodontal ligament at 24-26 days after birth when the occlusion of the first molars had been established. At the functional occlusion stage (60-80 days after birth), the Ruffini endings showing calretinin-immunoreactivity drastically increased in number and density, but less so than those positive for PGP 9.5-immunoreaction. The delayed expression of calretinin suggests that the function of the periodontal Ruffini endings is established after the completion of terminal formation because Ca2+, which binds to calcium binding proteins including calretinin with high affinity, plays an important role in mechano-electric transduction.

    DOI: 10.1679/aohc.62.57

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  • Effects of morphine on the distribution of Fos protein in the trigeminal subnucleus caudalis neurons during experimental tooth movement of the rat molar 査読

    Y Aihara, T Maeda, K Hanada, S Wakisaka

    BRAIN RESEARCH   819 ( 1-2 )   48 - 57   1999年2月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:ELSEVIER SCIENCE BV  

    The present study was undertaken to disclose temporal changes in the distribution of Fos-like immunoreactive (-IR) neurons in the trigeminal subnucleus caudalis (SpVc), one of the important relay nuclei for processing the nociceptive information from the oro-facial regions, following induction of experimental tooth movement in rat upper molars. Furthermore, the effect of morphine and naloxone on the levels of Fos-IR neurons in the SpVc was examined. The experimental tooth movement was induced by insertion of an elastic rubber between the first and second upper molars. In normal animals, Fos-IR neurons were rarely observed in the SpVc. immediately after insertion of the elastic band, the distribution of Fos-IR neurons was comparable to that observed in normal animals. The number of Fos-IR neurons increased significantly from 1 to 4 h following the induction of experimental tooth movement, reaching a maximum at 2 h, and then decreasing gradually. Most of the neurons were localized in the dorsomedial portion of the superficial layers of the ipsilateral SpVc near the obex, but a few were observed at the ventral portion of the SpVc. The neurons at the superficial layers and ventral portion of the contralateral SpVc also showed Foe-like immunoreactivity, but their numbers were significantly smaller than those on the ipsilateral side. Pretreatment with morphine (3 and 10 mg/kg, i.p.) significantly reduced the induction of Fos-IR neurons at the superficial layers of the ipsilateral SpVc in a dose-dependent manner, and its effect was antagonized by the subsequent treatment of naloxone (2 mg/kg, i.p.). Naloxone pretreatment enhanced the expression of Fos-IR neurons on the ipsilateral SpVc. The present results of a reduction of Fos-IR neurons by morphine pretreatment suggest that the induction of Fos-IR neurons may be due to the noxious stimulation caused by induction of experimental tooth movement. (C) 1999 Elsevier Science B.V. All rights reserved.

    DOI: 10.1016/S0006-8993(98)01323-7

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  • The distribution and ultrastructure of class II MHC-positive cells in human dental pulp 査読

    H Ohshima, T Maeda, Y Takano

    CELL AND TISSUE RESEARCH   295 ( 1 )   151 - 158   1999年1月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:SPRINGER VERLAG  

    The distribution and ultrastructure of class II major histocompatibility complex (MHC)-positive cells were investigated in human dental pulp, employing immunohistochemistry using an anti-human leukocyte antigen (HLA)DR-monoclonal antibody. HLA-DR-immunopositive cells, appearing spindle-like or dendritic in profile, were densely distributed throughout the dental pulp. Under the electron microscope, these cells exhibited various sizes of vesicles containing clear or opaque contents, multivesicular bodies and characteristic fine tubulovesicular structures in their cytoplasm. Some reactive cells possessed coated pits and vesicles including electron-dense materials, indicating an active endocytosis. At the periphery of the pulp tissue, the HLA-DR-immunopositive cells were predominantly situated in the subodontoblastic layer, with some located in the odontoblast layer and/or predentin and extending their cytoplasmic processes into the dentinal tubules. Cell processes of these cells occasionally made contact with several odontoblast processes in the same way as the nerve fibers in the predentin. These cells never contained the typical phagosomes frequently observed in the HLA-DR-immunoreactive macrophages in the subodontoblastic layer and the pulp core. The results suggest that the HLA-DR-immunopositive cells in the odontoblast layer and/or predentin have some regulatory function on the odontoblasts under physiological conditions, in addition to their involvement in the initial defense reaction after tooth injury.

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  • 乳歯埋伏の3症例 査読

    米山真理, 高木律男, 飯田明彦, 前田健康

    口腔科学会誌   48 ( 6 )   508 - 514   1999年

  • Re-innervation in the canine periodontal ligament of replanted teeth using an antibody to protein gene product 9.5: An immunohistochemical study 査読

    H. Yamada, T. Maeda, K. Hanada, Y. Takano

    Dental Traumatology   15 ( 5 )   221 - 234   1999年

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:Blackwell Munksgaard  

    The re-innervation process in the periodontal ligament of replanted canine teeth was examined by immunohistochemistry for protein gene product 9.5 (PGP 9.5), a general marker for neurons, and by electron microscopy. Within 1 week of replantation, the periodontal fibers had regenerated, filling the narrow spaces between the alveolar bone and the root surface around the cervical and apical regions. Near the root apex, however, no PGP 9.5-immunopositive nerve fibers were found in the regenerated periodontal ligament except for those in the alveolar half of the ligament. At 2 weeks after replantation, many nerve fibers positive for PGP 9.5 had ascended the periodontal ligament from the thick nerve bundles located near the root apex. Fine nerve endings showing complicated ramification were also present in the apical region. By 3 or 4 weeks after replantation, the vascular network was regenerated and principal periodontal fibers were re-established throughout the entire length of the periodontal ligament. The extensively ramified PGP 9.5-immunopositive structures had increased in thickness and density and showed characteristic tree-like profiles by 3 weeks. Electron microscopy confirmed that most of these structures were Ruffini-like endings, and demonstrated that such nerve terminals were almost regenerated by 4 weeks post-replantation. These results indicated that, in the periodontal ligament of replanted canine teeth, the regeneration of the nerve fibers including mechanosensory receptors first showed signs of regeneration by 2 weeks following tooth replantation and proceeded rapidly thereafter. Regeneration of the periodontal ligament including fiber architecture as well as vascular and neural elements was almost complete by 4 weeks after replantation.

    DOI: 10.1111/j.1600-9657.1999.tb00778.x

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  • Immunohistochemical localization of calbindin D28k during root formation of rat molar teeth 査読

    T. Onishi, T. Ooshima, S. Sobue, M. J. Tabata, T. Maeda, K. Kurisu, S. Wakisaka

    Cell and Tissue Research   297 ( 3 )   503 - 512   1999年

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    The present study was undertaken to examine the localization of calbindin D28k (CB)-like immunoreactivity (-LI) during the root formation of the rat molar. In the adult rat, CB-LI was detected in some of the cells of the epithelial rest of Malassez at the bifurcational region and in certain cells between the root dentin and cementum at the apical region. These cells had indented nuclei and many tonofilaments, and cementocytes lacked CB-LI, Moreover, CB-LI was observed in the periodontal fibroblasts in the alveolar half of the apical region. During root formation, the cells in the Hertwig's epithelial root sheath (HERS) lacked CB-LI, but most fragmented cells along the root surface began to express CB-LI when HERS was disrupted. Preodontoblasts and odontoblasts at the apical portion of the root also showed CB-LI. After the formation of cellular cementum, the CB-immunoreactive (-IR) cells were entrapped between the root dentin and cementum in the apical portion of the root. The number of CB-IR cells at the root surface decreased gradually, while that between the root dentin and cementum increased. The fibroblasts in the periodontal ligament began to express CB-LI after commencement of the occlusion, and the number and the staining intensity of CB-IR fibroblasts increased gradually with the passage of time. The present results suggest that CB may play an important role in the survival of the epithelial cells, in the cellular responses of periodontal fibroblasts against mechanical forces caused by the occlusion, and in the initial mineralization by the odontoblasts through the regulation of intracellular Ca2+ concentration.

    DOI: 10.1007/s004410051377

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  • Immunohistochemical localization of carbonic anhydrase isozyme II in the gustatory epithelium of the adult rat 査読

    Hiroshi Daikoku, Ichijiro Morisaki, Yuzo Ogawa, Takeyasu Maeda, Kojiro Kurisu, Satoshi Wakisaka

    Chemical Senses   24 ( 3 )   255 - 261   1999年

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    The distribution of carbonic anhydrase isozyme II (CA II)-like immunoreactivity (-LI) in the gustatory epithelium was examined in the adult rat. In the circumvallate and foliate papillae, CA II-LI was observed in the cytoplasm of the spindle-shaped taste bud cells, with weak immunoreaction in the surface of the gustatory epithelium. No neuronal elements displayed CA II-LI in these papillae. There was no apparent difference in the distribution pattern between the anterior and posterior portions of the foliate papillae. In immunoelectron microscopy, immunoreaction products for CA II were diffusely distributed in the entire cytoplasm of the taste bud cells having dense round granules at the periphery of the cells. No taste bud cells displaying CA II-LI were detected in the fungiform papillae, but a few thick nerve fibers displayed CA II-LI. In the taste buds of the palatal epithelium, neither taste bud cells nor neuronal elements exhibited CA II-LI. The present results indicate that CA II was localized in the type 1 cells designated as supporting cells in the taste buds located in the posterior lingual papillae of the adult animal.

    DOI: 10.1093/chemse/24.3.255

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  • Cytochrome oxidase activity in the enamel organ during amelogenesis in rat incisors 査読

    H Ohshima, T Maeda, Y Takano

    ANATOMICAL RECORD   252 ( 4 )   519 - 531   1998年12月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:WILEY-LISS  

    Cytochrome oxidase (CO), one of the membrane-bound mitochondrial enzymes involved in oxidative phosphorylation, reflects the functional activity of mitochondria. Mitochondria in the enamel organ show drastic changes in localization during amelogenesis (Smith. INSERM, 1984;125:273-282). In understanding the functional aspects of the enamel organ, it is essential that one knows the exact CO activity in the respective mitochondria. The present study examines the CO activity of mitochondria in the enamel organ of rat incisors throughout the various stages of amelogenesis using light and transmission electron microscopy. CO activity was examined histochemically according to Seligman et al. (J. Cell. Biol., 1968;38:1-14) in decalcified sections of the upper and lower incisors of the rat. In the secretory stage, half of the mitochondria in the ameloblasts accumulated in the infranuclear region were reactive for CO. Both the population and CO activity of the infranuclear mitochondria of ameloblasts decreased significantly in the later stage where the enamel matrix secretion was almost complete. The CO-reactive mitochondria in the cells of the stratum intermedium (SI) gradually increased in number throughout the secretory stage. In the maturation stage, the ameloblasts contained intensively CO-reactive giant mitochondria in the proximal region and regular sized ones in the distal cytoplasm that were mostly devoid of detectable CO reactivity. The proportion of CO-reactive mitochondria in the supranuclear region and the population of mitochondria in the infranuclear regions of the smooth-ended ameloblasts were significantly higher as compared with the respective values in the ruffle-ended ameloblasts. In the late stages of enamel maturation, ameloblasts containing a large number of ferritin-filled pigment vesicles possessed numerous CO-reactive mitochondria between those vesicles in the supranuclear region, implicating an active role of the ameloblasts in iron transfer into the maturing enamel. The papillary layer cells possessed numerous intensively CO-reactive mitochondria throughout the maturation stage. A stage-related variation in the localization of CO-reactive mitochondria in the enamel organ of rat incisors was quantitatively demonstrated. It is conceivable that maturation stage ameloblasts form a functional unit with the papillary layer cells, and operate in energy-requiring events such as active ion transport to, and water and matrix protein removal from the maturating enamel. A sign of such functional integrity among the types of the enamel organ cells (ameloblasts, cells of SI, cells of stellate reticulum, and outer enamel epithelial cells) cannot be seen in the secretory stage. The secretory ameloblasts may function in matrix formation and calcium regulation in a less cooperative manner with the other cells of the enamel organ as compared to the maturation stage ameloblasts. Anat. Rec. 252:519-531, 1998. (C) 1998 Wiley-Liss, Inc.

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  • Immunocytochemical demonstration of the synovial membrane in experimentally induced arthritis of the rat temporomandibular joint 査読

    K Nozawa-Inoue, R Takagi, T Kobayashi, Y Ohashi, T Maeda

    ARCHIVES OF HISTOLOGY AND CYTOLOGY   61 ( 5 )   451 - 466   1998年12月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:INT SOC HISTOLOGY & CYTOLOGY  

    The present study is first to report an experimental model of adjuvant-induced arthritis in the rat temporomandibular joint (TMJ). Arthritis was induced by simultaneous intradermal administrations of Freund's complete adjuvant, one at the parietal scalp and the other at the base of the tail. In this model, we demonstrated responses of the synovial membrane by immunocytochemistry using antibodies to OX6 and ED1 which recognize Ia antigen in MHC class II antigen-expressing cells and the macrophage/monocyte lineage, respectively. Three weeks after administration, no remarkable signs of inflammation were macroscopically recognizable in the TMJ, but microscopically the synovial membrane in the TMJ revealed marked changes such as enhanced vascularization and hemostasis in the sublining layer and a thickening in the synovial lining cell layer. Intense OX6-immunoreactivity was found in the synovial lining cells at lesions in the experimental group but not in the control group. Immunoelectron microscopy revealed that these OX6-immunopositive synovial lining cells developed dense cytoplasmic processes and numerous vacuoles and vesicles, resembling type A cells. Part of the type A cells also showed ED1-immunoreactivity. The expression of OX6 or ED1 immunoreactivity in the synovial lining cells might be involved in the initial immune responses in this arthritis model because the synovial membranes are exposed to the synovial fluids which have been believed to contain antigenic substances.

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  • Immunocytochemical detection of S-100 beta in the periodontal Ruffini endings in the rat incisor 査読

    K Nakakura-Ohshima, S Hayashi, Y Atsumi, S Wakisaka, K Nozawa-Inoue, T Maeda

    NEUROSCIENCE LETTERS   258 ( 3 )   163 - 166   1998年12月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:ELSEVIER SCI IRELAND LTD  

    Subcellular localization of S-100 protein, a kind of calcium binding proteins, was examined immunohistochemically in the Ruffini ending, a primary mechanoreceptor, in the periodontal ligament of the rat incisor. The periodontal ligament of the rat incisor was found to contain many S-100 beta- immunoreactive (-IR) structures but no S-100 alpha-IR elements. The S-100 beta-IR structures ramified extensively to form Ruffini endings and were frequently associated with round cells, the terminal Schwann cells, which also showed S-100 beta-like immunoreactivity. In many periodontal Ruffini endings, S-100 beta-IR products were recognized in the cytoplasm of Schwann cells, but not in the axoplasm. However, some axon terminals which had fewer or shorter axonal fingers, were filled with S-100 beta-IR products. The present findings indicated the existence of S-100 beta, not S-100 alpha, in axon terminals of the periodontal mechanoreceptive endings which were identified as type II Ruffini endings. (C) 1998 Elsevier Science Ireland ltd. All rights reserved.

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  • Three-dimensional direction and interrelationship of prisms in cuspal and cervical enamel of dog tooth 査読

    Y Hanaizumi, Y Kawano, H Ohshima, M Hoshino, K Takeuchi, T Maeda

    ANATOMICAL RECORD   252 ( 3 )   355 - 368   1998年11月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:WILEY-LISS  

    The three-dimensional architecture of enamel prisms was examined in cuspal enamel and compared with that in cervical enamel by light and electron microscopy as well as computer-assisted reconstruction using the developing enamel of several dog teeth.
    Dog tooth enamel consists of two groups of alternately arranged enamel prisms oriented in opposite sideward directions basically forming thick horizontal rings, partly branching off from the stem. Along a 8-10 enamel prism-wide group, the enamel prisms emerge in parallel tilting uniformly to the same sideward direction. In cervices, groups of enamel prisms are arranged nearly in parallel displaying a regular arrangement of prisms. Approaching the cusp of tooth, the groups of enamel prisms fuse to a concentric cusp-centered arrangement and the prisms exhibit no periodic arrangement as shown in the cervical enamel.
    It is suggested that the three-dimensional structure of enamel becomes complicated close to the cusp, contributing to the chewing stress of tooth. Anat. Rec. 252:355-368, 1998. (C) 1998 Wiley-Liss, Inc.

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  • Calretinin-like immunoreactivity in the regenerating periodontal Ruffini endings of the rat incisor following injury to the inferior alveolar nerve 査読

    K Hiroshima, T Maeda, K Hanada, S Wakisaka

    BRAIN RESEARCH   807 ( 1-2 )   218 - 221   1998年10月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:ELSEVIER SCIENCE BV  

    Regeneration of calretinin (CR)-like immunoreactive (IR) nerve fibers was investigated in the periodontal ligament of the rat lower incisor following resection of the inferior alveolar nerve (IAN). Ln addition, the degeneration and regeneration processes of periodontal nerve fibers were examined by immunohistochemistry for protein gene product 9.5 (PGP 9.5), a general neuronal marker. in normal animals, the periodontal nerve fibers showing PGP 9.5-like immunoreactivity (LI) formed either periodontal Ruffini endings with expanded arborization and thin free nerve endings in the alveolar half of the ligament. Thick CR-IR nerve fibers also appeared in a dendritic fashion in the same region, but thin CR-IR nerve fibers were rarely observed. Five days following resection of the IAN, a major population of PGP 9.5-IR and all CR-IR nerve fibers disappeared except for some thin PGP 9.5-IR nerves in the periodontal Ligament. Regenerated PGP 9.5-IR nerve fibers appear:ed around 7 days following resection, in contrast to a very small number of regenerated CR-IR nerve fibers. Around 14-21 days following resection, the number and terminal morphology of regenerated PGP 9.5-IR nerve fibers were comparable to those observed in normal animals, but the number of regenerated CR-IR nerve fibers was still smaller than that of normal animals. The number of regenerated CR-IR nerve fibers increased to return to normal by 56 days following injury. The delay of expression of CR-LI in the regenerated periodontal Ruffmi endings suggests that functional recovery of periodontal Ruffini endings occurred after the completion of the regeneration of periodontal nerve fibers. (C) 1998 Elsevier Science B.V. All rights reserved.

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  • Experimental chronic infection induced in mice by Actinomyces israelii entrapped in alginate gel 査読

    MAA Moral, H Ohshima, T Maeda, E Hoshino

    ARCHIVES OF ORAL BIOLOGY   43 ( 6 )   485 - 496   1998年6月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:PERGAMON-ELSEVIER SCIENCE LTD  

    Tissue responses to experimentally induced actinomycotic lesions were investigated in mice by both light and transmission electron microscopy. Micro-organisms of Actinomyces israelii were entrapped in alginate gel and injected into the subcutaneous tissue over the periosteum of the mouse cranium. One day after the injection (initial stage), a non-stained amorphous structure was located in the core of the lesion, corresponding to the injected gel with bacteria. Numerous neutrophils surrounded the core region and phagocytized the injected complex actively. At days 3-7 (intermediate stage), the lesion became well developed. The core structure became eosinophilic and separated to form island-like structures. No lesion was recognized in the control group (gel without bacteria) until day 14. After 30 days (late stage), the lesions displayed more static features, similar to the "sulphur granules" characteristic of actinomycotic lesions. At the late stage, foamy cells increased in number and took the place of neutrophils in the alginate islands. By transmission electron microscopy these foamy cells were seen to be filled with lysosomal Vesicles containing electron-dense foreign material. Thus, these cells appeared to be macrophages that had phagocytized degenerated neutrophils containing bacteria. Along with the active phagocytosis by foamy cells that progressed in the late stage, a collagenous capsule became conspicuous and separated the lesion from the intact tissue. The bacteria remained in the gel islands until at least day 60, although they considerably decreased in number with time. Serum IgG antibody titres began to rise within 24 h of the injection: reached a peak concentration at day 14 and remained a significantly high (p &lt; 0.01, vs 0 time) until day 120. These results suggest that this animal model is useful for inducing experimental chronic infectious lesions. (C) 1998 Elsevier Science Ltd. All rights reserved.

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  • A histological study on tissue responses to titanium implantation in rat maxilla: The process of epithelial regeneration and bone reaction 査読

    N Fujii, H Kusakari, T Maeda

    JOURNAL OF PERIODONTOLOGY   69 ( 4 )   485 - 495   1998年4月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:AMER ACAD PERIODONTOLOGY  

    THE PRESENT STUDY AIMED TO ESTABLISH a titanium implantation model using rat maxillae as well as demonstrate the chronological tissue responses to implantation. Pure titanium implants were inserted in the upper first molar extraction sites of Wistar rats 1 month after tooth extraction. The animals were sacrificed at 1 to 30 days postimplantation, and prepared tissue specimens were processed for light microscopy. The removal of implants from tissue blocks was done using 2 methods: mechanical removal or a cryofracture technique. In the early stages, peri-implant tissues showed severe damage to the oral epithelium and collagen bundles with significant inflammatory cell infiltration. The peri-implant epithelium grew apically along the implant by 10 days postimplantation, and regenerated to show a similar feature of junctional epithelium seen in normal rats at 15 days postimplantation, at which time no signs of inflammation were observed. The regenerated collagen bundles in the connective tissue were arranged circumferentially to the implants in the horizontal sections. New bone formation first appeared around the implants at 5 days postimplantation, covering the entire perimeter of implants by 30 days postimplantation. Scanning electron microscopic observations of the surface texture of the removed implants suggest the probability of an adhesive mechanism between the implants and the peri-implant epithelium and/or the alveolar bone. These findings indicate that this experimental model is useful for detailed analysis of peri-implant tissue because of its easy implantation procedure.

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  • 歯根膜ルフィニ神経終末の形態学的基盤 査読

    前田 健康, 大島 勇人

    解剖学雑誌   73 ( 2 )   119 - 134   1998年4月

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    記述言語:日本語   出版者・発行元:(一社)日本解剖学会  

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  • Alterations in ultrastructural localization of growth-associated protein-43 (GAP-43) in periodontal Ruffini endings of rat molars during experimental tooth movement 査読

    H Kobayashi, K Ochi, Saito, I, K Hanada, T Maeda

    JOURNAL OF DENTAL RESEARCH   77 ( 3 )   503 - 517   1998年3月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:AMER ASSOC DENTAL RESEARCH  

    It is known that orthodontic forces induce discomfort and/or abnormal sensation after application of an orthodontic appliance in patients, suggesting the adaptation of periodontal neural elements to environmental changes. However, no morphological data have been provided. The present study investigated, by immunoelectron microscopy, the localization of growth-associated protein-43 (GAP-43) in periodontal Ruffini endings in rat molars during experimental tooth movement. In the untreated control group, immunoelectron microscopy demonstrated that GAP-43-like immunoreactivity in the Ruffini endings was confined to the Schwann sheaths around the axon terminals, and was in neither the cell bodies of terminal Schwann cells nor the axon terminals themselves. Immunoelectron microscopic observation revealed alterations in the localization of GAP-43-like immunoreactivity in the periodontal Ruffini endings during experimental tooth movement. After 1 day of treatment, the cell bodies of the terminal Schwann cells associated with Ruffini endings appeared to contain immunoreaction products for GAP-43, and retained GAP-43-like immunoreactivity during tooth movement. From 5 to 7 days, a major population of the axoplasm of the periodontal Ruffini endings, which was immunonegative in control, filled the GAP-43 immunoreactions, showing a tendency to decrease in number later, and disappeared completely at 14 days. These findings suggest that orthodontic forces easily induce the remodeling of the mechanoreceptive Ruffini endings as well as the active tissue remodeling in a close relationship. Since the ultrastructural localization of GAP-43-like immunoreactivity was drastically changed in the Ruffini endings during tooth movement, GAP-43 functions as one of the key molecules in the remodeling of mechanoreceptive Ruffini endings during tooth movement.

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  • Class II MHC antigen-expressing cells in the pulp tissue of human deciduous teeth prior to shedding 査読

    N Kannari, H Ohshima, T Maeda, T Noda, Y Takano

    ARCHIVES OF HISTOLOGY AND CYTOLOGY   61 ( 1 )   1 - 15   1998年3月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:INT SOC HISTOLOGY & CYTOLOGY  

    The distribution and ultrastructure of the class II major histocompatibility complex (MHC) antigen-expressing cells in the pulp tissue of human deciduous teeth during the process of physiological root resorption was surveyed by histochemical and immunocytochemical methods using an anti-human leukocyte antigen (HLA)-DR-monoclonal antibody, Dental pulp was found to contain numerous HLA-DR-positive cells of various shapes; those showing a dendritic appearance were located mainly in the periphery of the pulp tissue, associated closely with the odontoblasts. The immunopositive cells sometimes extended their cytoplasmic processes into the dentinal tubules and increased in number in the areas affected by dental caries, attrition or restorative procedures, implicating their role in immunosurveillance.
    The immunopositive cells were located consistently at the pulp-dentin border during the stage of active resorption, adjacent to the preodontoclasts or odontoclasts, and covered the exposed dentin surface after the detachment of the odontoclasts until the onset of cementum formation, These data suggest that the HLA-DR-immunopositive cells in the coronal pulp of human deciduous teeth play an inductive role in the differentiation, migration and/or activation of the odontoclasts and cementoblast-like cells during the stages of tooth resorption.

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  • Growth-associated protein-43 (GAP-43) in the regenerating periodontal Ruffini endings of the rat incisor following injury to the inferior alveolar nerve 査読

    SH Youn, T Maeda, K Kurisu, S Wakisaka

    BRAIN RESEARCH   787 ( 1 )   41 - 48   1998年3月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:ELSEVIER SCIENCE BV  

    Alterations in the levels of growth-associated protein 43 (GAP-43)-like immunoreactivity (-LI) were examined in the lingual periodontal ligament of the rat incisor following two types of injury (resection and crush) to the inferior alveolar nerve (IAN). In normal animals, GAP-43-like immunoreactive (IR) structures were observed as tree-like ramifications in the alveolar half of the lingual periodontal ligament of incisors. Under immunoelectron microscopy, GAP-43-LI appeared in the Schwann sheaths associated with periodontal Ruffini endings; neither cell bodies of the terminal Schwann cells nor axonal profiles showed CAP-43-LI. During regeneration of the periodontal Ruffini endings following resection of the IAN, GAP-43-LI appeared in the cytoplasm of the terminal Schwann cell bodies and axoplasm of the terminals. The distribution of GAP-43-LI in the Ruffini endings returned to almost normal levels on days 28 and 56 following the injury. The changes in the distribution of GAP-43-LI following the crush injury were similar to those following resection; however, expression of GAP-43-LI was slightly higher for the entire experimental period compared with the resection. The transient expression of GAP-43 in the terminal Schwann cells and axonal profiles of the periodontal Ruffini endings following nerve injury suggests that GAP-43 is closely associated with axon-schwann cells interactions during regeneration. (C) 1998 Elsevier Science B.V.

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  • Adenosquamous carcinoma of the tongue - Report of a case with histochemical, immunohistochemical, and ultrastructural study and review of the literature 査読

    K Izumi, T Nakajima, T Maeda, J Cheng, T Saku

    ORAL SURGERY ORAL MEDICINE ORAL PATHOLOGY ORAL RADIOLOGY AND ENDODONTICS   85 ( 2 )   178 - 184   1998年2月

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    記述言語:英語   出版者・発行元:MOSBY-YEAR BOOK INC  

    A rare case of oral adenosquamous carcinoma in a 78-year-old woman is reported. The tumor occurred in her tongue and metastasized to the submandibular and cervical lymph node;. Histologically, the tumor showed invasive growth involving the submucosal and muscle lavers. Its solid carcinomatous nests exhibited ductal differentiation in the deeper aspects and squamous differentiation toward the surface. Histochemical examination revealed an accumulation of acid mucopolysaccharide in the ductal luminal and the ductal cells were immunohistochemically positive for carcinoembryonic antigen, epithelial membrane antigen, cancer antigen 15-3 and Ulex europaeus agglutinin I. Ultrastructurally, tonotibrils, desmosomes and numerous cytoplasmic processes were common features of the tumor cells. In addition, true glandular structures and pseudocysts were seen in areas. Clinical features of 13 adenosquamous carcinomas in the literature were analyzed.

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  • Carbonic anhydrase isozyme II immunoreactivity in the mechanoreceptive Ruffini endings of the periodontal ligament in rat incisor 査読

    K Ochi, S Wakisaka, SH Youn, K Hanada, T Maeda

    BRAIN RESEARCH   779 ( 1-2 )   276 - 279   1998年1月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:ELSEVIER SCIENCE BV  

    The present study describes the distribution of carbonic anhydrase isozyme II (CA II) in the lingual periodontal ligament of the rat incisor. Some thick nerve fibers in the nerve bundle displayed CA II-like immunoreactivity (LI) as well as non-neuronal elements such as osteoclasts. At the alveolar half of the lingual periodontal ligament of the incisor, thick CA II-like immunoreactive (-IR) nerve fibers showed a tree-like raminification, but thin and beaded CA II-IR nerve fibers were rare. Under the electron microscope, CA II-LI were diffusely localized in the axoplasm of the axon terminals surrounded by Schwann sheaths which were immunonegative for CA II. The cell bodies of the terminal Schwann cells associated with the periodontal Ruffini endings did not exhibit CA II-LI. The present immunohistochemical evidence indicates that CA II may participate in the regulation of the intra-neuronal ion in the periodontal Ruffini endings which are thought to be in a state of high neuronal activity. (C) 1998 Elsevier Science B.V.

    DOI: 10.1016/S0006-8993(97)01085-8

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  • Innervation of the periodontal ligament in the alligatorid Caiman crocodilius 査読

    Osamu Tadokoro, Hiroyuki Mishima, Takeyasu Maeda, Yukishige Kozawa

    European Journal of Oral Sciences   106 ( 1 )   519 - 523   1998年1月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    The mode of development and structure of crocodilian teeth and periodontium parallels that of mammals, but the teeth are continuously replaced throughout the lifetime of those animals. In this report, the innervation and fibres of the crocodilian periodontal ligament were examined using histology, immunohistochemistry for S-100 protein and transmission electron microscopy. Crocodilian periodontal ligaments had the following characteristics: (1) horizontal fibres, which connect the alveolar bone to the root cementum and (2) longitudinal fibres, which ran parallel to the tooth axis, with nerves and blood vessels in the middle layer of the ligament. The apex of root and tooth germs were both embedded in thick circular fibres. S-100 protein was detected in neural elements including terminal portions which were densely distributed in the periodontal ligament and dental follicle. The S-100 positive neural elements formed a periodontal plexus. We found two types of nerve endings
    free endings and simple encapsulated corpuscles as described in mammals. The presence of such nerve endings in caiman suggests that these teeth, in addition to having a biting function, may also act as highly sensitive sensory organs. © Eur J Oral Sci, 1998.

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  • Immunohistochemical localization of pleiotrophin and midkine in the lingual epithelium of the adult rat 査読

    Satoshi Wakisaka, Makoto J. Tabata, Takeyasu Maeda, Kazumasa Matsumoto, Akio Wanaka, Hisako Muramatsu, Takashi Muramatsu, Kojiro Kurisu

    Archives of Histology and Cytology   61 ( 5 )   475 - 480   1998年

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:Japan Society of Histological Documentation  

    Distribution of two heparin-binding molecules, pleiotrophin (PTN) and midkine (MK), was investigated by immunohistochemistry in the lingual epithelium of the adult rat. In the lingual epithelium, both PTN- and MK- like immunoreactivities were observed in its basal cell layers, showing a mesh-like appearance. These molecules were also found along the surface of the taste bud cells
    an intense immunoreaction was detected at the base of the taste buds in the circumvallate and foliate papillae. At the electron microscope level, the immunoreactive products were localized on the cell surface and basement membrane at the base of the taste buds. The immunoreactivity for PTN was distributed more diffusely than that for MK. It was suggested that these molecules may be involved in the differentiation and maintenance of taste bud cells, possibly via their trophic effect upon approaching nerves.

    DOI: 10.1679/aohc.61.475

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  • Morphological basis on periodontal ruffini endings 査読

    Takeyasu Maeda, Hayato Ohshima

    Acta Anatomica Nipponica   73 ( 2 )   131 - 134   1998年

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    記述言語:日本語   掲載種別:研究論文(学術雑誌)   出版者・発行元:Japanese Association of Anatomists  

    The periodontal ligament has a rich sensory nerve supply which serves as a sensory apparatus in addition to tooth support. The periodontal ligament contains nociceptors and low-threshold mechanoreceptors. Stimuli applied to teeth evoke various oral reflexes, which make smooth mastication possible via the periodontal mechanoreceptors. Recent morphological and physiological studies have revealed that Ruffini endings, categorized as low-threshold slowly adapting type II (SA II), are essential mechanoreceptors in the periodontal ligament. The periodontal Ruffini endings are ultrastructurally characterized by expanded axon terminals filled with a number of mitochondria and terminal or lamellar Schwann cells. The axon terminals of the periodontal Ruffini endings have finger-like projections, i.e. axonal spines, extending into the surrounding tissue to detect the deformation of collagen fibers. As histochemical marker enzymes for the periodontal Ruffini endings, the axon terminals and terminal Schwann cells are reactive for cytochrome oxidase activity and both acid phosphatase activity and non-specific cholinesterase activity, respectively. Many experimental studies also have revealed that periodontal Ruffini endings have high potential for neuroplasticity, confirmed by intense immunoreactivity for p75NGFR and GAP-43. Mechanical stimuli due to tooth eruption and occlusion might be a prerequisite for the differentiation and maturation of the periodontal Ruffini endings. Further investigations are needed for clarifying the involvement of growth factors and the molecular mechanism of the development and regeneration processes of the Ruffini endings.

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  • Distribution of calcium ions at the interface between resin bonding materials and tooth dentin. Use of commercially available adhesive systems 査読

    Y Hanaizumi, T Maeda, Y Takano

    JOURNAL OF ELECTRON MICROSCOPY   47 ( 3 )   227 - 241   1998年

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:OXFORD UNIV PRESS  

    It has been proposed that calcium ions play a key role in chemical (chelate) binding between the adhesive resin and dentin surface. However, no data is available concerning how calcium ions are distributed at the binding sites. The aim of this study is to demonstrate calcium ions at the resin-dentin interface by means of X-ray microanalysis and calcium ion-sensitive histochemical staining. The dentin surface in human teeth was ground by use of 240 grit silicon carbide abrasive paper under running water and treated with the dentin-primer and adhesive resin in Clearfil Liner Bond System or IMPERVA Bond System according to the manufacturer's instructions. After removing dentin matrix and isolating adhesive resin by the KOH-digestion method, one half of the samples were processed for scanning electron microscopy. The rest were embedded in Epon 812 and processed either for glyoxal bis (2-hpdroxyanil) (GBHA) staining or transmission electron microscopy combined with X-ray microanalysis. Transmission electron microscopy revealed Ca-phosphate deposits at the bottom of the resin-impregnated layer. The adhesive resin above the resin-impregnated layer was amorphous and showed no precipitates of Ca-phosphate. GBHA displayed intense calcium reactions throughout the resin-impregnated layer and also moderate ones in the 10 mu m (Clearfil Liner Bond System) or 30 mu m (IMPERVA Bonding System) thick boundary zone of the adhesive resin as well as in the resin tags. These data are the first to offer a distinct localization of calcium ions within the adhesive resin at the dentin-resin interface.

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  • Immunohistochemical observation of growth-associated protein 43 (GAP-43) in the developing circumvallate papilla of the rat 査読

    S. Wakisaka, H. Daikoku, Y. Miyawaki, S. H. Youn, T. Maeda, K. Kurisu

    Cell and Tissue Research   293 ( 3 )   499 - 507   1998年

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    The distribution and development of growth-associated protein 43 (GAP-43)-like immunoreactivity (-LI) in the rat circumvallate papilla (CVP) were compared to those of protein gene product 9.5 (PGP 9.5)-LI. In the adult, thick GAP-43-like immunoreactive (-IR) structures gathered densely in the subgemmal region. Some of these further penetrated the apical epithelium and trench wall epithelium. At least two types of GAP-43-IR structures were recognized
    taste bud-related and non-gustatory GAP-43-IR neural elements. Immunoelectron microscopy revealed that GAP-43-LI was localized predominantly in the Schwann cells, and a few axons displayed GAP-43-LI in the lamina propria. In the trench epithelium, GAP-43-LI was detected in the cytoplasmic side of the axonal membrane. Some intragemmal GAP-43-IR axons made synaptic-like contacts with taste bud cells. At least four developmental stages were defined on the basis of the changes in distribution of GAP-43-LI. In stage I [embryonic day (E) 16-17] GAP-43-IR structures accumulated at the lamina propria just beneath the newly-formed circumvallate papilla. In stage II (E18-19) GAP-43-IR nerve fibers began to penetrate the apical epithelium. In stage III [E20-postnatal day (P) 0] GAP-43-IR nerve fibers first appeared in the trench wall epithelium. Penetration of GAP-IR nerve fibers occurred in the inner trench wall epithelium first, and then in the outer trench wall epithelium. In stage IV (P1-) the distribution of GAP-43-LI was similar to that observed in the adult
    but the density of GAP-43-LI was much higher than in adults. PGP 9.5-LI showed a similar distribution pattern to that of GAP-43-LI, except for round-shaped cells in the apical epithelium at the late embryonic stages, and in taste bud cells and intralingual ganglionic cells which lacked GAP-43-LI. The similarities in distribution patterns of GAP-43-LI and PGP 9.5-LI during the development and mature circumvallate papilla suggest that GAP-43 may be a key neuronal molecule for induction and maintenance of the taste buds.

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  • Calbindin D28k-like immunoreactivity in the developing and regenerating circumvallate papilla of the rat 査読

    Y. Miyawaki, I. Morisaki, M. J. Tabata, T. Maeda, K. Kurisu, S. Wakisaka

    Cell and Tissue Research   291 ( 1 )   81 - 90   1998年

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    The distribution of calbindin D28k (CB)-like immunoreactivity (-LI) in the circumvallate papilla (CVP) was examined during development and regeneration following bilateral crush injury to the glossopharyngeal nerve in the rat. In the adult CVP, CB-like immunoreactive (-IR) nerve fibers were observed in the subgemmal region and some penetrated into the taste buds. CB-LI was also detected in the cytoplasm of the spindle-shaped gustatory cells in the lower half of the trench epithelium, which contained numerous synaptic vesicles and bundles of intermediate filaments. These CB-IR gustatory cells made synapse-like contacts with CB-IR nerve terminals. Some CB-IR nerve terminals made contacts with the gustatory cells negative for CB-LI. At least three developmental stages were defined with regard to the developmental changes in the distribution of CB-LI: (1) Stage I (embryonic day (E) 18-postnatal day (P)5): CB-IR nerve fibers appeared in the lamina propria just beneath the newly-formed CVP at E18, but the gustatory epithelium of the CVP contained no CB-IR structures. Taste buds with taste pores appeared at P1. (2) Stage II (P5-10): thin CB-IR nerve fibers began entering the trench epithelium, but no CB-IR cells were observed. (3) Stage III (P10-adult): in addition to the intragemmal and perigemmal CB-IR nerve fibers, very few CB-IR cells appeared in the taste buds around P10, and their numbers increased progressively. The changes in the distribution of taste buds and CB-LI following glossopharyngeal nerve injury were similar to those observed during development. On post-operative day (PO) 4, the taste buds and CB-IR cells decreased markedly in number. These CB-IR cells be came round in shape, and the number of CB-IR nerve fibers decreased markedly. On PO8, both taste buds and CB-IR cells disappeared completely. The regenerated taste buds were first observed on PO12, increased rapidly in number by PO20, and increased slowly thereafter. CB-IR nerve fibers accumulated at the subgemmal region and began penetrating into the trench wall epithelium around PO16. CB-IR cells appeared between PO20 and PO24, and their numbers increased progressively and reached the normal level on PO40. The topographical localizations of the taste buds and CB-IR cells during development and regeneration were comparable to those of normal animals. The delay of the time courses for appearance of CB-IR nerve fibers and CB-IR cells compared to the appearance of taste buds during development and regeneration suggests that CB in the gustatory epithelium may participate in the survival of the taste bud cells rather than in the induction of the taste buds.

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  • Scanning and transmission electron microscopy of Ruffini endings in the periodontal ligament of rat incisors 査読

    Hiromi Takahashi-Iwanaga, Takeyasu Maeda, Kazuhiro Abe

    Journal of Comparative Neurology   389 ( 1 )   177 - 184   1997年12月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    The Ruffini organ is an arborized axon ending categorized as a low- threshold stretch receptor. We have previously shown that the lingual periodontal ligament of rat incisors is densely innervated with Ruffini endings. In the present study, fine structures in the surface of the periodontal Ruffini endings and their topographical relationship with the surrounding collagen fibers were observed by a combination of scanning and transmission electron microscopy to analyze the mechanism of the stretch reception. The entire length of the branches of the Ruffini endings, excepting their terminal portions, corresponded well with those depicted by previous investigators in the following points: (1) their cylindrical appearance covered by Schwann cell processes
    (2) the presence of numerous axon fingers protruding through gaps in the Schwann sheath and
    (3) their isolation from collagen fibers by multilayered basal lamina. On the other hand, tips of the axon branches-together with their Schwann sheaths-became attenuated and projected into tight bundles of collagen, indicating their susceptibility to mechanical deformations of the surrounding tissue. Margins of the axon terminals were conspicuously ruffled with long tongue-like projections of Schwann cells. The Schwann cell tongues twined around collagen bundles in their distal portions, and associated closely with fine axon projections in their proximal portions, suggesting their involvement in the mechanical transmission of stimuli to axon terminals.

    DOI: 10.1002/(SICI)1096-9861(19971208)389:1<177::AID-CNE13>3.0.CO;2-I

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  • Immunocytochemical demonstration of beta(1)-subunit of Na+/K+-ATPase in the mechanoreceptive Ruffini-like endings of the rat incisor ligament 査読

    K Ochi, K Hanada, SH Youn, S Wakisaka, T Maeda

    ARCHIVES OF ORAL BIOLOGY   42 ( 10-11 )   779 - 782   1997年10月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:PERGAMON-ELSEVIER SCIENCE LTD  

    The localization of one of the isoforms of Na+/K+-ATPase, the beta(1)-subunit, was investigated in the periodontal Ruffini endings of rat incisors by light-and electron-microscopic immunocytochemistry. Immunoreactivity for the rat beta(1)-subunit followed the pattern of dendritic terminal arborization in the alveolar half of the lingual periodontal ligament. Ultrastructurally, the reaction products were localized in dilatations of axons, possibly the terminals of Ruffini-like endings in the periodontal ligament. No immunoreactivity was seen in Schwann cells. The immunostaining results support the view that the beta(1)-subunit of Na+/K+-ATPase is the predominant isoform in sensory neurones, and that this protein is a useful marker for periodontal Ruffini-like endings. (C) 1997 Elsevier Science Ltd.

    DOI: 10.1016/S0003-9969(97)00097-6

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  • Calbindin D28k-like immunoreactive nerve fibres in the predentine of rat molar teeth 査読

    Y Miyawaki, SH Youn, K Ochi, T Maeda, K Kurisu, S Wakisaka

    ARCHIVES OF ORAL BIOLOGY   42 ( 10-11 )   773 - 777   1997年10月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:PERGAMON-ELSEVIER SCIENCE LTD  

    Immunoelectron-microscopy was applied to reveal the existence of nerve fibres and terminals showing calbindin D28k (CB)-like immunoreactivity (IR) in the rat molar tooth pulp. In the root pulp, thick, smooth-surfaced CB-IR nerve fibres were in bundles accompanying the blood vessels. In the coronal pulp, the fibres arborized repeatedly and extensively. CB-IR nerve fibres had a predominantly thick, smooth-surfaced appearance, though parts appeared thin and beaded. Occasionally some thin. varicose CB-IR nerve fibres ran along the odontoblasts, penetrating into the predentine alongside the dentinal tubules. They could be traced for approx. 10-20 mu m into the predentine from the pulp-predentine border. Immunoelectron-microscopy revealed that only some of the nerve terminals in the predentine showed CB-IR, and that predentinal CB-IR nerve terminals were located close to the odontoblast processes. No synaptic structures were observed between them. The presence of CB-IR nerve terminals in the predentine suggests that many, if not all, CB-IR nerve fibres could be nociceptors. The CB could be involved in Ca homeostasis during the activation of nociceptors. (C) 1997 Elsevier Science Ltd.

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  • Calretinin-like immunoreactivity in the Ruffini endings, slowly adapting mechanoreceptors, of the periodontal ligament of the rat incisor 査読

    K Ochi, S Wakisaka, SH Youn, K Hanada, T Maeda

    BRAIN RESEARCH   769 ( 1 )   183 - 187   1997年9月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:ELSEVIER SCIENCE BV  

    The distribution and ultrastructural localization of calretinin (CR)-like immunoreactivity (-LI) were investigated in the lingual periodontal ligament of rat incisors. Some thick nerve fibers within the nerve bundle displayed CR-LI; these CR-Like immunoreactive (-IR) nerve fibers entered the alveolar half of the lingual periodontal ligament of the incisor where dendritic terminal arborization was exhibited. Thin and beaded CR-IR nerve fibers were rarely observed in the periodontal ligament. Observations of adjacent sections immunostained with protein gene-product 9.5 (PGP 9.5) revealed that most, if not all, PGP 9.5-IR nerve terminals showing a dendritic arborization expressed CR-LI. Immunoelectron microscopic observations showed that electron-opaque immunoreaction products were localized in the axoplasm of the axon terminals, except for the mitochondria, which were surrounded by Schwann sheaths and multiple-layered basal lamina. Neither cell bodies, the cytoplasmic extension of terminal Schwann cells, nor other cellular elements such as periodontal fibroblasts exhibited CR-LI. The present findings suggest that Ruffini endings, an essential mechanoreceptor in the periodontal ligament and categorized as a slowly adapting mechanoreceptor, express CR-LI, and that CR may participate in the Ca2+ homeostasis against external stimuli in the periodontal Ruffini endings. (C) 1997 Elsevier Science B.V.

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  • Expression of TrkB-like immunoreactivity in non-neural cells of rat periodontal ligament 査読

    K Ochi, Saito, I, K Hanada, T Maeda

    ARCHIVES OF ORAL BIOLOGY   42 ( 6 )   455 - 464   1997年6月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:PERGAMON-ELSEVIER SCIENCE LTD  

    The Trk family, a group of high-affinity neurotrophin receptors, is divided into three subtypes, TrkA, TrkB and TrkC. These were originally found in neural elements, and are involved in neural development, maintenance and survival. Recent studies have shown that non-neural cells in vitro also express mRNA encoding some neurotrophin receptors. In our preliminary study, TrkB-like immunoreactivity (LI) was found in the various non-neural cells in the rat periodontal ligament. The present study was undertaken to clarify which cell types express Trk-LI, in particular two types of TrkB-LI, in the periodontal ligament of mature rats? using an immunocytochemical technique with polyclonal antibodies. Intense full-length TrkB-LI was clearly recognized in non-neural cells such as fibroblasts, osteoclasts, odontoclasts and cementoblasts as well as in neural elements. Relatively large cells with many cytoplasmic processes were also frequently immunopositive for full-length TrkB. Immunocytochemistry for TrkB[TK-], a truncated type, also demonstrated a similar immunostaining pattern to that of full-length TrkB in non-neural periodontal cells, and intense positive reactions in endothelial cells. Some non-neural cells were positive for TrkA and TrkC. These findings suggest that neurotrophic factors, the ligands of the Trk family, have certain effects on the proliferation and/or differentiation of non-neural cells, as well as on their neurotrophic functions. (C) 1997 Elsevier Science Ltd.

    DOI: 10.1016/S0003-9969(97)00030-7

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  • Immunohistochemical localization of calbindin D28k in the periodontal Ruffini endings of rat incisors 査読

    K Ochi, S Wakisaka, SH Youn, K Hanada, T Maeda

    NEUROSCIENCE LETTERS   228 ( 3 )   195 - 198   1997年6月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:ELSEVIER SCI IRELAND LTD  

    It was examined whether calbindin D28k (CB) might be located in the rat incisor periodontal Ruffini ending, an essential mechanoreceptor in periodontal ligament, by light- and electron-microscopic immunohistochemistry. Some thick nerve fibers showing CB-like immunoreactivity (LI) entered the lingual half of the periodontal ligament of the incisor and showed the dendritic terminal arborization. Electron-dense immunoreaction products indicating CB-LI were distributed diffusely in axoplasm of the axon terminals, no mitochondria, however, were not labeled. Neither cell bodies nor cytoplasmic extensions of the terminal Schwann cells exhibited CB-LI. CB was presumed to be involved in the maintenance of Ca2+ homeostasis in the mechano-electric transduction in mechanoreceptors in the periodontal ligament. (C) 1997 Elsevier Science Ireland Ltd.

    DOI: 10.1016/S0304-3940(97)00398-4

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  • Subpopulation of Class II MHC Antigen-expressing Cells in the Rat Incisor Pulp as Shown by Acid Phosphatase Histochemistry

    OHSHIMA Hayato, MAEDA Takeyasu, TAKANO Yoshiro

    Dentistry in Japan   33   8 - 14   1997年3月

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  • 象牙質・歯髄複合体の病態生理に関する研究 査読

    下野 正基, 井上 孝, 橋本 貞充, 高橋 和人, 須田 英明, 前田 健康, 脇坂 聡

    日本歯科医学会誌   16   77 - 81   1997年3月

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    記述言語:日本語   出版者・発行元:日本歯科医学会  

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  • A histological investigation on tissue responses to titanium implants in cortical bone of the rat femur 査読

    A Ohtsu, H Kusakari, T Maeda, Y Takano

    JOURNAL OF PERIODONTOLOGY   68 ( 3 )   270 - 283   1997年3月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:AMER ACAD PERIODONTOLOGY  

    IMPLANT MATERIALS ARE PLACED under various sites-including cortical bone, spongy bane, and bone marrow-at the same time according to the depth at implantation. Although cortical bone is an important site for the prognosis of implantation, detailed reports on tissue responses to implantation have been meager. The present study aims to reveal tissue responses to pure titanium implantation in rat femoris cortical bone. The rats received titanium bars surgically in their femurs and were sacrificed 1 day to 40 weeks post-implantation. The prepared tissue specimens were processed for light and transmission electron microscopy (TEM). Further histochemical detections were performed. One day post-implantation, empty osteocytic lacunae indicating degeneration of osteocytes were found in pre-existing cortical bone around the implant. Such pre-existing bone was replaced by new bone, but remained in part even 40 weeks post-implantation. Light microscopy showed that direct contact between the implant and new bone was identified 12 weeks post-implantation, Chronological and ultrastructural observation showed that new bone deposition appeared to proceed toward the implant, and that the intervening layer at the interface was derived from the degenerated debris of multinucleated giant cells and/or osteoblasts. Furthermore, it seemed that the width of intervening layer varied in relation to the distance from the blood vessels. The cells showing tartrate resistant acid phosphatase activity possessed cytological features of osteoclasts under TEM; they were frequently observed in perivascular sites near the implants even after osseointegration, suggesting that bone remodeling took place steadily around the implant.

    DOI: 10.1902/jop.1997.68.3.270

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  • Three-dimensional arrangement of enamel prisms and their relation to the formation of Hunter-Schreger bands in dog tooth 査読

    Y Hanaizumi, T Maeda, Y Takano

    CELL AND TISSUE RESEARCH   286 ( 1 )   103 - 114   1996年10月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:SPRINGER  

    The three-dimensional architecture of enamel prisms and their relationship to Hunter-Schreger bands were examined in the developing enamel of several dog teeth by light and electron microscopy, and computer-assisted reconstruction. Sections were prepared from a single demineralized tooth germ. Longitudinal semithin sections parallel to the meridian of the tooth showed parazones and diazones of the Hunter-Schreger bands in alternate rows at equal intervals. On sections vertical to the tooth crown through the middle region of parazone or diazone, a row of parallel prisms were angulated with the largest tilting angle being 55 degrees to the enamel-dentin junction: running in opposite directions in the respective zones. Tangential sections parallel to the enamel-dentin junction showed numerous belt-like zones arranged perpendicular to the meridian of the tooth. Each belt-like zone consisted of a group of enamel prisms oriented in the same direction, those in the neighboring zones being oriented in an opposite direction. The densely stained boundaries between the adjacent belt-like zones corresponded to the interface between parazone and diazone. Computer-reconstructed enamel prisms in the adjacent two zones were oriented in the opposite sidewards direction with occasional confluence and divergence. Scanning electron-microscopic observation of the developing enamel surface exposed by dissolution of the enamel organ revealed band-like arrangements of groups of pits encasing the Tomes' processes of secretory ameloblasts. The secretory faces of the pits inclined uniformly in the same sidewards direction, with those in the neighboring groups, in the opposite direction.

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  • Three-dimensional arrangement of enamel prisms and their relation to the formation of Hunter-Schreger bands in dog tooth 査読

    Y Hanaizumi, T Maeda, Y Takano

    CELL AND TISSUE RESEARCH   286 ( 1 )   103 - 114   1996年10月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:SPRINGER  

    The three-dimensional architecture of enamel prisms and their relationship to Hunter-Schreger bands were examined in the developing enamel of several dog teeth by light and electron microscopy, and computer-assisted reconstruction. Sections were prepared from a single demineralized tooth germ. Longitudinal semithin sections parallel to the meridian of the tooth showed parazones and diazones of the Hunter-Schreger bands in alternate rows at equal intervals. On sections vertical to the tooth crown through the middle region of parazone or diazone, a row of parallel prisms were angulated with the largest tilting angle being 55 degrees to the enamel-dentin junction: running in opposite directions in the respective zones. Tangential sections parallel to the enamel-dentin junction showed numerous belt-like zones arranged perpendicular to the meridian of the tooth. Each belt-like zone consisted of a group of enamel prisms oriented in the same direction, those in the neighboring zones being oriented in an opposite direction. The densely stained boundaries between the adjacent belt-like zones corresponded to the interface between parazone and diazone. Computer-reconstructed enamel prisms in the adjacent two zones were oriented in the opposite sidewards direction with occasional confluence and divergence. Scanning electron-microscopic observation of the developing enamel surface exposed by dissolution of the enamel organ revealed band-like arrangements of groups of pits encasing the Tomes' processes of secretory ameloblasts. The secretory faces of the pits inclined uniformly in the same sidewards direction, with those in the neighboring groups, in the opposite direction.

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  • Immunoelectron microscopic study on neuropeptide Y in the periodontal ligament of the incisor following peripheral nerve injury to the inferior alveolar nerve in the rat 査読

    S Wakisaka, SH Youn, T Maeda, K Kurisu

    BRAIN RESEARCH   729 ( 2 )   259 - 263   1996年8月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:ELSEVIER SCIENCE BV  

    Immunoelectron microscopic analysis was carried out to examine whether neuropeptide Y (NPY)-like immunoreactivity (-LI) is localized in mechanoreceptors in the lingual periodontal ligament of the rat incisor following peripheral nerve injury to the inferior alveolar nerve (IAN). In the lingual periodontal ligament of normal animals, no NPY-like immunoreactive (-IR) primary afferents were observed, except for a very few sympathetic perivascular nerve fibers which showed NPY-LI. Fourteen days following chronic constriction injury to the LAN combined with sympathectomy of the superior cervical ganglion, thick NPY-IR nerve fibers showing tree-like raminifications were detected in the shear zone between the tooth-related part and alveolus-related part as well as in the alveolus-related part. Immunoelectron microscopy revealed that expanded NPY-IR nerve terminals were covered with several Schwann sheaths and that a part of the axoplasm expanded to the surrounding tissues. These ultrastructural features of NPY-IR structures were identical to those of periodontal Ruffini endings, categorized as slowly adapting mechanoreceptors. Thick (6-8 mu m in diameter) NPY-IR axons were also observed without any apparent myelin sheath. The present results provide further evidence that NPY is closely associated with thick axons, probably myelinated nerves and Ruffini endings, following peripheral nerve injury.

    DOI: 10.1016/S0006-8993(96)00570-7

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  • Different localizations of growth-associated protein (GAP-43) in mechanoreceptors and free nerve endings of adult rat periodontal ligament, dental pulp and skin 査読

    T Maeda, MR Byers

    ARCHIVES OF HISTOLOGY AND CYTOLOGY   59 ( 3 )   291 - 304   1996年8月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:INT SOC HISTOLOGY & CYTOLOGY  

    Distributions of growth-associated protein-43 (GAP-43) in the periodontal ligament and dental pulp of adult rats were studied by light and electron microscopy. The mature periodontal ligament and dental pulp contained numerous GAP-43-positive neural elements, comprising periodontal Ruffini endings and thin nerve fibers, but expression patterns differed among the kinds of nerves, In the periodontal ligament of rat molars, immunoelectron microscopy revealed that GAP-43 like immunoreactivity in the Ruffini ending, an essential mechanoreceptor, was confined to the Schwann sheaths around the axon terminals and was not in the axon terminals themselves, unlike free endings that revealed axonal GAP-43, However, the lamellar Schwann cells associated with the cutaneous receptors did not exhibit any GAP-43 like immunoreactivity though they were intensely reactive for low affinity nerve growth factor receptor (p75-NGFR), a marker for lamellar Schwann cells in mechanoreceptors, The characteristically uniform expression of GAP-43 in the Schwann lamellae that surround the Ruffini mechanoreceptors of rat molar ligament suggests that Schwann cells are involved in the GAP-43 mediated plasticity of these receptors. On the other hand, the pulpal nerves were filled with the reaction products in their axonal spaces, suggesting the potential for neuronal plasticity during normal function and after tooth injury.

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  • Neuropeptide Y-like immunoreactive primary afferents in the periodontal tissues following dental injury in the rat 査読

    S Wakisaka, SH Youn, T Maeda, K Kurisu

    REGULATORY PEPTIDES   63 ( 2-3 )   163 - 169   1996年7月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:ELSEVIER SCIENCE BV  

    The distribution of neuropeptide Y (NPY)-like immunoreactive (-IR) nerve fibers in the periodontal tissues following dental injury to the rat maxillary first molar was examined with a combination of dental injury and surgical sympathectomy of the superior cervical ganglion (SCGx). In normal animals, NPY-IR nerve fibers were observed around the blood vessels of the trigeminal ganglion, dental pulp and periodontal tissues. These nerve fibers completely disappeared following SCGx. Fourteen days following dental injury of the maxillary first molar combined with SCGx, a small number NPY-IR cells was observed in the dorsal to middle portion of the maxillary division of the trigeminal ganglion. These were mostly medium- to large-sized cells with a mean +/- SD cross-sectional area of 541.4 +/- 239.3 mu m(2). Approx. 50% of these cells had the cross-sectional areas between 400-600 mu m(2) In the periodontal tissues of injured first molar, thick NPY-IR nerve fibers showing an irregular appearance were detected in the apical region. Immunoelectron microscopy showed that most NPY-IR nerve fibers near the lower half of the injured periodontal ligament had an axonal diameter of approx. 7-8 mu m, and lacked apparent myelin sheaths. Near NPY-IR nerve fibers, many macrophages with phagosomes containing debris of the myelin sheaths were observed. At the oral epithelium covering the injured roots of the maxillary first molar, thick NPY-IR nerve fibers were recognizable and some penetrated the epithelium. No NPY-IR nerve fibers were observed in the dental pulp or periodontal tissues in second and third molars, and ultrastructural views of nerve fibers were almost intact following combined SCGx and dental injury to the first molar. The present results indicate that NPY-IR primary afferents appeared in the periodontal tissues following dental injury, and that NPY may be closely associated with the regeneration process of injured primary afferents.

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  • Responses of periodontal nerve terminals to experimentally induced occlusal trauma in rat molars: An immunohistochemical study using PGP 9.5 antibody 査読

    T Sodeyama, T Maeda, Y Takano, K Hara

    JOURNAL OF PERIODONTAL RESEARCH   31 ( 4 )   235 - 248   1996年5月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:MUNKSGAARD INT PUBL LTD  

    The response of periodontal nerves to experimentally induced occlusal trauma in rat molars was assessed by immunohistochemistry for protein gene product 9.5 (PGP 9.5) at light and electron microscopic levels, and by computerized image analysis. The occlusal surface on the left upper first molar of 8-wk-old male Wistar rats was raised approximately 1 mm under ether anaesthesia. The rats were perfusion-fixed on d 1, 2, 3, 4, and 7 after bite-raising and then decalcified for 2-3 wk. Frozen sagittal cryostat sections were stained by the avidin-biotin complex method. By the second day after bite-raising many Ruffini endings were swollen and their outline unclear at the light microscopic level. Transmission electron microscopy disclosed PGP 9.5 reaction products within Ruffini endings that had unusually long cytoplasmic projections extending through enlarged slits of the Schwann sheaths and also diffuse extracellular PGP 9.5-immunoreactivity near the Ruffini endings. From d 2 to 4, thin nerve fibres on the pressure side of the per iodontal ligament were orientated irregularly and had a prominent beaded appearance. An increase in beaded nerve terminals occurred at d 2-4 post elevation, and decreased later. These results suggest that occlusal trauma induces specific changes in the distribution and shape of nerve terminals in the periodontal ligament.

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  • Dendritic Cells: A Novel Cellular Component of the Rat Incisor Enamel Organ in the Late Stages of Enamel Maturation. 査読

    H. Ohshima, O. Sato, I. Kawahara, T. Maeda, Y. Takano

    Adv. Dent. Res.   10 ( 2 )   94 - 104   1996年

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

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  • Responses of class II MHC antigen-expressing cells to cavity preparation 査読

    H Ohshima, Y Takano, O Sato, Kawahara, I, T Maeda

    DENTIN/PULP COMPLEX   316 - 318   1996年

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    記述言語:英語   掲載種別:研究論文(国際会議プロシーディングス)   出版者・発行元:QUINTESSENCE PUBL CO INC  

    Responses of class II major histocompatibility complex (MHC) antigen-expressing cells to cavity preparation were investigated both in the human premolars and molars and the upper first molars of rats. In the human control teeth, class II MHC antigen-expressing cells were widely distributed throughout the dental pulp. The HLA-DR-immunopositive cells in the periphery of the pulp tissue showed dendritic appearance, and some of them being located in the predentin, extending their cytoplasmic processes into the dental tubules. In the control rat molars, OX6-immunopositive cells were located in the subodontoblastic layer and not found in the predentin. Many OX6-immunopositive cells accumulated along the pulp-dentin border 12-24 hours after cavity preparation and extended their cytoplasmic processes into the exposed dentinal tubules. In the human teeth, numerous neutrophil leucocytes migrated deep into the exposed dentinal tubules 24 hours after drilling, while most of the immunopositive cells were situated beneath the exudative lesion. The present results suggested the difference between class II MHC antigen-expressing cells in the human teeth and rat molars in chronological responses to foreign stimuli. It is also confirmed that numerous leucocytes migrate deep into the exposed dentinal tubules when destructive changes are induced in odontoblasts by cavity preparation in the human teeth.

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  • POSTNATAL-DEVELOPMENT OF PERIODONTAL RUFFINI ENDINGS IN RAT INCISORS - AN IMMUNOELECTRON MICROSCOPIC STUDY USING PROTEIN-GENE-PRODUCT-9.5 (PGP-9.5) ANTIBODY 査読

    K NAKAKURAOHSHIMA, T MAEDA, H OHSHIMA, T NODA, Y TAKANO

    JOURNAL OF COMPARATIVE NEUROLOGY   362 ( 4 )   551 - 564   1995年11月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:WILEY-LISS  

    Postnatal development of Ruffini endings was ultrastructurally investigated in the upper incisors of the rat from 1 day to 60 days after birth by means of protein gene product 9.5 (PGP 9.5) immunocytochemistry. The immunostaining with PGP 9.5 antibody clearly demonstrated chronological alterations of the distribution and ultrastructure of the Ruffini endings during postnatal development. At 1 day after birth, the PGP 9.5-positive nerve terminals contained a few mitochondria and vesicles immunonegative for PGP 9.5. Dendritic terminals appeared at 4 days after birth, with a small number of expanded or bulbous portions. These expanded portions possessed morphological features similar to those of the growth cone: several mitochondria and various kinds of vesicles. Typical Ruffini endings with dendritic ramification and expanded portions appeared 7-11 days after birth. At this stage, parts of the axon terminals extended through the slits of Schwann cell covering and formed finger-like projections called axonal spines. These Ruffini endings increased dramatically in number after 24-26 days and were identical in density and morphology to those seen in adult rats. After the commencement of the occlusion between the incisors, the number of large mitochondria increased, in contrast to the decrease of the vesicles in the axon terminals. Moreover, the axonal spines increased both in number and in length. Thus, the periodontal nerve endings showed stage-specific morphological features intimately related in timing to tooth eruption and occlusion. Functional stimuli possibly contribute to the final differentiation and maturation of the periodontal Ruffini endings. (C) 1995 Wiley-Liss, Inc.

    DOI: 10.1002/cne.903620409

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  • PRIMARY LEIOMYOSARCOMA OF THE MAXILLA WITH REGIONAL LYMPH-NODE METASTASIS - REPORT OF A CASE AND REVIEW OF THE LITERATURE 査読

    K IZUMI, T MAEDA, J CHENG, T SAKU

    ORAL SURGERY ORAL MEDICINE ORAL PATHOLOGY ORAL RADIOLOGY AND ENDODONTICS   80 ( 3 )   310 - 319   1995年9月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:MOSBY-YEAR BOOK INC  

    A rare case of oral leiomyosarcoma diagnosed with the aid of immunohistochemical and electron microscopic examinations together with a review of the literature are reported. The patient was a 70-year-old Japanese man. The primary tumor involved the maxillary gingiva and bone and metastasized to the cervical lymph nodes. On histologic examination the tumor showed invasive growth into the maxillary bone. It was composed of interlacing fascicles of spindle-shaped cells with eosinophilic cytoplasm and elongated, blunt-ended nuclei. The tumor formed extensive metastatic foci in the cervical lymph nodes. On immunohistochemical examination most of the tumor cells were positive for desmin, smooth muscle-specific actin, and myosin. The ultrastructural characteristics of the tumor cells were abundant microfilaments, pinocytotic vesicles, and basement membrane formation. The findings were indicative of a tumor demonstrating myogenic differentiation. A review of the literature during the past 50 years disclosed a total of 60 oral leiomyosarcomas, including our case.

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  • Responses of immunocompetent cells to cavity preparation in rat molars: An immunohistochemical study using OX6-monoclonal antibody 査読

    H Ohshima, O Sato, Kawahara, I, T Maeda, Y Takano

    CONNECTIVE TISSUE RESEARCH   32 ( 1-4 )   303 - 311   1995年

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:GORDON BREACH SCI PUBL LTD  

    Responses of immunocompetent cells, especially class II major histocompatibility complex (MHC) antigen-expressing cells, were investigated after cavity preparation in the erupted upper first molar teeth of rats, by immunohistochemistry using OX6-monoclonal antibody.
    In control teeth, OX6-immunopositive cells were predominantly located beneath the odontoblast layer in the dental pulp. Cavity preparation caused an acute edematous reaction between the injured odontoblasts and predentin, and most of OX6-immunopositive cells in the affected site shifted away from the pulp-dentin border. After 12-24 hours, many OX6-immunopositive cells accumulated along the pulp-dentin border and extended their cytoplasmic processes into the exposed dentinal tubules. After 72 hours, newly differentiated odontoblasts replaced the degenerated odontoblasts, and few OX6-immunopositive cells remained along the pulp-dentin border.
    Our data suggest that some of the class II MHC antigen-expressing cells in the dental pulp participate in the initial defense reaction and presumably serve asa biological sensor for the external stimuli arriving through the exposed dentinal tubules.

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  • Ultrastructural and immunocytochemical characterization of a gingival neurofibroma. 査読

    K. Izumi, T. Nakajima, T. Maeda, T. Saku

    Assian J. Oral Maxillofac. Surg.   7 ( 1 )   3 - 11   1995年

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

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  • THE RELATIONSHIP BETWEEN ODONTOBLASTS AND IMMUNOCOMPETENT CELLS DURING DENTINOGENESIS IN RAT INCISORS - AN IMMUNOHISTOCHEMICAL STUDY USING OX6-MONOCLONAL ANTIBODY 査読

    H OHSHIMA, KAWAHARA, I, T MAEDA, Y TAKANO

    ARCHIVES OF HISTOLOGY AND CYTOLOGY   57 ( 5 )   435 - 447   1994年12月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:INT SOC HISTOLOGY & CYTOLOGY  

    The relationship between odontoblasts and class II major histocompatibility complex (MHC) antigen-expressing cells in the process of dentinogenesis was studied in rat lower incisors, employing immunohistochemistry using OX6 monoclonal antibody, The dental pulp contained numerous OX6-immunopositive cells that varied in morphology from dendritic to spindle under physiological conditions, Under the electron microscope, these immunopositive cells shared common cytoplasmic features, i.e,, multivesicular bodies and characteristic fine tubulovesicular structures in their cytoplasm. At the early stage of dentinogenesis, OX6-immunopositive cells, presumably of the immature type, were located in the subodontoblastic layer, During active dentin formation, the OXG-immunopositive cells increased in number and appeared in the odontoblast layer, associating intimately with fenestrated capillaries situated close to the predentin, These cells showed a dendritic appearance and possessed various sizes of multivesicular bodies and characteristic fine tubulovesicular structures, but never contained typical phagosomes, On the other hand, immunopositive macrophages characterized by typical phagosomes tended to occupy the central portion of the pulp,
    The results suggest that most, if not all, OX6-immunopositive cells situated deep in the odontoblast layer are dendritic cells playing a role in the defense system of the dental pulp against antigenic molecules arriving from the circulation via the fenestrated capillaries, The increasing number of OXG-immunopositive or immunonegative macrophages appearing near the incisal end of the tooth is thought to be involved in the elimination of degenerated odontoblasts.

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  • EVIDENCE FOR THE EXISTENCE OF INTRAEPITHELIAL NERVE-ENDINGS IN THE JUNCTIONAL EPITHELIUM OF RAT MOLARS - AN IMMUNOHISTOCHEMICAL STUDY USING PROTEIN GENE-PRODUCT-9.5 (PGP-9.5) ANTIBODY 査読

    T MAEDA, T SODEYAMA, K HARA, Y TAKANO

    JOURNAL OF PERIODONTAL RESEARCH   29 ( 6 )   377 - 385   1994年11月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:MUNKSGAARD INT PUBL LTD  

    Innervation of the junctional epithelium was investigated in rat molars by means of immunohistochemistry for protein gene product 9.5 (PGP 9.5) at light and electron microscopic levels. In comparison with our previous study on same tissues using neurofilament protein (NFP)-antibody, the PGP 9.5-immunostaining further disclosed numerous nerve fibers in the gingiva of rat molars and revealed the existence of a well-developed plexus of PGP 9.5-positive nerve fibers. The interproximal portion also contained numerous nerve fibers.
    Observation of horizontal sections revealed a denser innervation toward the inner junctional gingival epithelium than the outer marginal epithelium. The nerve fibers, beaded in appearance and extending from the nerve bundles in the lamina propria, penetrated into the junctional epithelial layer and were distributed throughout the junctional epithelium, with some nerves being located near the epithelial surface. Non-neuronal cells showing PGP 9.5-immunoreactivity were absent in the junctional epithelium. In immunoelectron microscopy, the axoplasm of nerves in the gingiva was filled with electron-dense reaction products of PGP 9.5, except for the cell organellae. The nerve fibers were devoid of Schwann cell investment and terminated among the epithelial cells in the junctional epithelium, frequently beneath the epithelial surface. The intraepithelial nerve endings contained various kinds of Vesicles including large-cored ones, supporting the presence of peptidergic innervation shown by previous studies. These findings confirmed the usefulness of PGP 9.5-immunohistochemistry for the identification of delicated nerve fibers in dental tissue, and suggested the dense network of nerve fibers that may serve as sensory receptor and other functions in the junctional epithelium.

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  • DENSE INNERVATION OF HUMAN RADICULAR DENTAL-PULP AS REVEALED BY IMMUNOCYTOCHEMISTRY FOR PROTEIN GENE-PRODUCT-9.5 査読

    T MAEDA, S HONMA, Y TAKANO

    ARCHIVES OF ORAL BIOLOGY   39 ( 7 )   563 - 568   1994年7月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:PERGAMON-ELSEVIER SCIENCE LTD  

    Protein gene-product 9.5 (PGP 9.5) is a novel neurone-specific protein. At light- and electron-microscopic levels, human radicular dental pulp was heavily innervated by PGP 9.5 nerve fibres. Thick nerve bundles showing intense PGP 9.5 immunoreactivity ascended in the centre of the pulp, with some nerve fibres extending from the trunk towards the peripheral pulp at regular intervals. However, the fibres did not form a plexus (of Raschkow) beneath the odontoblast cell layer. The PGP 9.5-positive nerve fibres penetrated into the predentine and dentine beyond the odontoblast cell layer where some nerves terminated. In the predentine, PGP 9.5-positive nerve fibres were densely distributed. In tangential sections along the pulpodentinal border, the nerve fibres ran in contact with odontoblast processes. Immunoelectron microscopy revealed that all nerve fibres were immunoreactive for PGP 9.5 in the radicular predentine and dentine, and terminated in contact with the odontoblast cell processes. The distribution pattern and terminal formation of predentinal and dentinal nerves were identical to those of coronal pulp, as reported previously. The dense innervation in radicular dental pulp, overlooked in previous reports, might be responsible for dentine hypersensitivity.

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  • ラット顎関節の神経支配 抗PGP9.5血清を用いた免疫組織学的研究

    小林龍彰, 大橋靖, 前田健康, 高野吉郎

    日本口腔科学会雑誌   43 ( 3 )   369 - 385   1994年7月

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  • ラット顎関節の神経支配 −PGP 9.5 血清を用いた免疫組織化学的研究− 査読

    小林龍彰, 大橋 靖, 前田健康, 高野吉郎

    口科誌   43 ( 3 )   369 - 385   1994年

  • ALTERATION OF NERVE GROWTH FACTOR-RECEPTOR EXPRESSION IN THE PERIODONTAL-LIGAMENT OF THE RAT DURING EXPERIMENTAL TOOTH MOVEMENT 査読

    SAITO, I, K HANADA, T MAEDA

    ARCHIVES OF ORAL BIOLOGY   38 ( 11 )   923 - 929   1993年11月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:PERGAMON-ELSEVIER SCIENCE LTD  

    This pilot study deals with the initial responses of neural elements showing nerve growth factor-receptor (NGFR) immunoreactivity in the periodontal ligament of rats to orthodontic forces. The animals were killed at intervals of 1, 2, 6, 12 h and 1, 3, 5, 7 days after the insertion of elastic bands between the maxillary first and second molars. Serial frozen sections. prepared from each animal, were processed immunohistochemically to demonstrate NGFR, after which the periodontal ligament of the mesial root of the maxillary second molar was examined microscopically. In control sections, NGFR-positive neural elements were predominantly distributed at the apex of the bony socket on the distal side of the periodontal ligament. After 1 h of tooth movement. NGFR positively-stained nerve fibres tended to disappear slightly in both the intermediate and coronal regions of the distal periodontal ligament. By the third day of tooth movement, the periodontal ligament nerve fibres showed more intense NGFR-immunoreactivity; thick, positively stained nerve fibres were recognized on the distal side in which active bone remodelling was occurring, and a few of these fibres were densely distributed around blood vessels or near Howship's lacunae. The distribution of NGFR-positive neural elements on the mesial side increased at 5 days of tooth movement. Finally, at 7 days of tooth movement, staining intensity for NGFR appeared to decrease. These findings demonstrate that an alteration in the distribution and the intensity of immunoreactive staining for NGFR in the periodontal ligament is associated with the bone remodelling induced by orthodontic tooth movement.

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  • POSTNATAL-DEVELOPMENT OF PERIODONTAL INNERVATION IN RAT INCISORS - AN IMMUNOHISTOCHEMICAL STUDY USING PROTEIN GENE-PRODUCT 9.5 ANTIBODY 査読

    K NAKAKURAOHSHIMA, T MAEDA, O SATO, Y TAKANO

    ARCHIVES OF HISTOLOGY AND CYTOLOGY   56 ( 4 )   385 - 398   1993年10月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:INT SOC HISTOLOGY & CYTOLOGY  

    The postnatal development of innervation in the lingual periodontal ligament of rat incisors was investigated by immunohistochemistry using an antibody against protein gene product 9.5 (PGP 9.5), a neuron-specific protein.
    Immunostaining with the antibody enabled the clear demonstration of chronological alteration in the distribution and maturation of nerve terminals after birth. At 1 day after birth, thick nerve fibers showing PGP 9.5-immunoreactivity first appeared in the loose connective tissue corresponding to the future periodontal ligament, without forming an apparent terminal structure. At 4 days, two portions became distinguishable in the lingual periodontal ligament: alveolus- and tooth-related parts being comparable to those in the adult ligament. The immunoreactive nerve fibers increased in density and appeared to extend more incisally. Those restricted to the alveolus-related part showed ramifications with a small number of expanded or bulbous portions, and terminated among the periodontal fibers. Characteristic dendritic and expanded terminals, similar to Ruffini endings in the adult ligaments, began to occur in the lingual periodontal ligament at 7 days after birth and gradually increased in number until day 11 when the incisal edges emerged in the oral cavity. At approximately 15 days after birth, nerve endings possessing morphological features similar to Ruffini endings in adult rats were recognizable in the entire length of the periodontal ligament. After the eruption of first molars, the periodontal Ruffini endings drastically increased in number, and by the time the occlusion between the first molars was established, became identical in distribution and density to those seen in the lingual periodontal ligament of adult rat incisors.
    These findings suggest that the mechanical stimuli due to tooth eruption and occlusion might be a prerequisite for the final differentiation and maturation of the periodontal Ruffini endings.

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  • HISTOCHEMICAL-DEMONSTRATION OF ACID-PHOSPHATASE-ACTIVITY IN TERMINAL SCHWANN-CELLS ASSOCIATED WITH RUFFINI ENDINGS IN THE PERIODONTAL-LIGAMENT OF RAT INCISORS 査読

    T MAEDA, O SATO, KAWAHARA, I, Y TAKANO

    ARCHIVES OF ORAL BIOLOGY   38 ( 7 )   611 - 617   1993年7月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:PERGAMON-ELSEVIER SCIENCE LTD  

    Histochemical staining for acid phosphatase, a marker for lysosomal elements, distinguished rounded, intensely reactive cells from less reactive fibroblasts and osteoblasts in the lingual periodontal ligament. The highly reactive cells were located exclusively in the alveolar half of the ligament. Double staining for acid phosphatase and S-100 protein confirmed that these reactive cells were identical with the terminal Schwann cells associated with periodontal Ruffini endings. Electron microscopically, reaction products for acid phosphatase were observed in the lysosomes and Golgi apparatus in the paranuclear cytoplasm of the terminal Schwann cells. As the terminal Schwann cells associated with the Ruffini endings are assumed to be capable of synthesizing exportable proteins, acid phosphatase in this type of cell may be involved in the processing of macromolecules in synthetic and/or secretory pathways.

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  • HISTOCHEMICAL-LOCALIZATION AND X-RAY-MICROANALYSIS OF CALCIUM IN THE RAT SUBMANDIBULAR-GLAND - DEMONSTRATION OF POSSIBLE SITES FOR MICROLITH INDUCTION 査読

    Y TAKANO, Y SATO, H OHSHIMA, T MAEDA, KAWAHARA, I, NOGUCHI, I

    ARCHIVES OF HISTOLOGY AND CYTOLOGY   56 ( 2 )   177 - 184   1993年6月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:INT SOC HISTOLOGY & CYTOLOGY  

    Rapidly frozen and freeze-substituted submandibular glands of young female rats were embedded in Epon and processed for histochemical demonstration of calcium with the glyoxal bis (2-hydroxyanil) (GBHA) staining method. GBHA staining of thick Epon sections revealed discrete calcium reactions of moderate intensity in practically every secretory granule but not in other compartments of the acinar cells. The saliva in the excretory duct was also reactive with GBHA and showed a drastic decrease in staining intensity toward the distal segments of excretory ducts with larger diameters. In addition, the duct saliva contained numerous tiny particles that were highly GBHA reactive. Stromal cells and cells lining the excretory duct were totally free of reactions. In the acinar cells, X-ray analysis detected distinct peaks for calcium in secretory granules and smaller ones in the Golgi apparatus, while they were undetectable in the rough surfaced endoplasmic reticulum (RER), implicating post-RER calcium loading in the secretory pathway. Electron-dense deposits in the duct saliva showed distinct peaks both for calcium and phosphorus, though these appeared in the acinar secretory granules and other cytoplasmic regions lacked phosphorus.
    Our observations thus demonstrated physiologcal calcium in the intra- as well as extracellular compartments of the submandibular gland, and further confirmed drastic changes in chemical composition along the synthetic and secretory pathways of the saliva, by both histochemical and X-ray microanalytical methods. GBHA staining of calcium combined with X-ray microanalysis is useful for an evaluation of the physiology and histo-pathological changes of the salivary glands associated with initial phases of microliths as well as sialoliths formation.

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  • CYTOCHROME-OXIDASE ACTIVITY AS A MARKER FOR PERIODONTAL SENSORY RECEPTORS IN THE RAT 査読

    T MAEDA, O SATO, Y TAKANO

    ARCHIVES OF ORAL BIOLOGY   38 ( 3 )   255 - 259   1993年3月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:PERGAMON-ELSEVIER SCIENCE LTD  

    Cytochrome oxidase activity was explored histochemically in axon terminals of periodontal Ruffini endings of rat upper incisors at both the light- and electron-microscopic levels. Staining clearly demonstrated ramified structures in the alveolar half of the periodontal ligament. These structures resembled the profiles of the axon terminals of the periodontal Ruffini ending previously demonstrated by an immunohistochemical method for neurofilament protein. Histochemically at the electron-microscopic level, the ramified structures were identified as true Ruffini endings in which each axon terminal was filled with reactive mitochondria. Two types of mitochondria were distinguished with respect to the localization pattern of reaction products; almost all mitochondria were positive for cytochrome oxidase activity, with only a few negative. As the enzyme activity did not decrease after demineralization, the findings suggest that cytochrome oxidase is a useful marker enzyme for demonstrating sensory receptors in the periodontal ligament. Histochemical methods for cytochrome oxidase may contribute to the light- and electron-microscopic morphological analysis of periodontal sensory receptors.

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  • SINUSOIDAL VESSELS IN THE PERIODONTAL-LIGAMENT OF HAMSTER INCISORS - THEIR DISTRIBUTION, STRUCTURE AND POSSIBLE FUNCTION 査読

    K KANNARI, T MAEDA, Y TAKANO

    ARCHIVES OF HISTOLOGY AND CYTOLOGY   56 ( 1 )   91 - 99   1993年3月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:INT SOC HISTOLOGY & CYTOLOGY  

    Vascular architecture in the periodontal ligament of hamster incisors was investigated by use of vascular casts under a scanning electron microscope (SEM). In addition to ordinary nutrient blood vessels, anastomosing vessels of large caliber developed, surrounding the incisor. From their characteristic configuration, these vessels were regarded as ''sinusoids''. The plexus of sinusoidal vessels was connected with capillaries in the papillary layer of the enamel organ at the labial periodontal ligament, and with veins penetrating into the alveolar bone on the lingual side. Transmission electron microscopic (TEM) observation showed that the sinusoidal wall was composed of only a thin layer of endothelial cells, lacking a smooth muscular element, and surrounded by densely arranged collagen fibers. Although the frequent association of Ruffini-type nerve endings with siunsoidal vessels was noted, neither direct contact nor specialized structures between these was recognizable. A possible function of the periodontal sinusoids is discussed on the basis of their distribution and ultrastructural evidence.

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  • インプラント周囲組織の経時的変化 −特に神経線維の動態について− 査読

    澤田雅仁, 草刈 玄, 佐藤 修, 前田健康, 高野吉郎

    補綴誌   37 ( 1 )   144 - 158   1993年2月

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    記述言語:日本語  

    DOI: 10.2186/jjps.37.144

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  • GBHA staining methods: Its application to demonstration of cytoplasmic calcium stores in some exocrine and endocrine glands. 査読

    Y. Takano, H. Ohshima T. Maeda, Y. Sato

    J. Hard Tiss. Biol.   2 ( 2 )   1 - 23   1993年

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

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  • Three-dimensional Structure of Dog Sertoli Cells: A Computer-Aided Reconstruction from Serial Semi-Thin Sections 査読

    Koichi Kannari, Takeyasu Maeda, Yoshiro Takano

    Archives of Histology and Cytology   56 ( 1 )   65 - 73   1993年

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    Vascular architecture in the periodontal ligament of hamster incisors was investigated by use of vascular casts under a scanning electron microscope (SEM). In addition to ordinary nutrient blood vessels, anastomosing vessels of large caliber developed, sur-rounding the incisor. From their characteristic con-figuration, these vessels were regarded as sinusoids. The plexus of sinusoidal vessels was connected with capillaries in the papillary layer of the enamel organ at the labial periodontal ligament, and with veins pene-trating into the alveolar bone on the lingual side. Transmission electron microscopic (TEM) observation showed that the sinusoidal wall was composed of only a thin layer of endothelial cells, lacking a smooth muscu-lar element, and surrounded by densely arranged colla-gen fibers. Although the frequent association of Ruffini-type nerve endings with siunsoidal vessels was noted, neither direct contact nor specialized structures be-tween these was recognizable. A possible function of the periodontal sinusoids is discussed on the basis of their distribution and ultrastructural evidence. © 1993, International Society of Histology and Cytology. All rights reserved.

    DOI: 10.1679/aohc.56.65

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  • IMMUNOELECTRON MICROSCOPIC OBSERVATION OF CALCITONIN GENE-RELATED PEPTIDE (CGRP)-POSITIVE NERVES IN THE DENTAL-PULP OF RAT MOLARS 査読

    O SATO, H TAKEUCHIMAENO, T MAEDA, Y TAKANO

    ARCHIVES OF HISTOLOGY AND CYTOLOGY   55 ( 5 )   561 - 568   1992年12月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:INT SOC HISTOLOGY & CYTOLOGY  

    The ultrastructure of nerves containing immunoreactivity for calcitonin gene-related peptide (CGRP) was investigated in the dental pulp of rat molars. The immunoreactivity was recognized predominantly in unmyelinated nerve fibers, and sparsely in a few myelinated fibers. It was localized throughout the axoplasm, as well as in the large cored vesicles. Small clear vesicles and mitochondria were free of the immunoreaction. The CGRP-immunoreactive nerves were frequently observed to terminate, being devoid of Schwann cell investment, in the vicinity of blood vessels in the coronal pulp, suggesting that CGRP may be involved in the regulation of pulpal blood flow. Moreover, CGRP-immunoreactive axon terminals containing numerous small clear vesicles, a few large cored vesicles and mitochondria were recognized in contact with the cell bodies of odontoblasts and their processes in the dentinal tubules. Although specialized synaptic ultrastructures were not recognizable, a functional association of CGRP nerves and odontoblasts was suggested. Thus, CGRP in the dental pulp appears to have multiple functions, including vascular regulation and sensory transduction.

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  • 実験的ハムスター舌癌組織における神経線維の動態:抗NFP,S‐100蛋白抗体を用いた免疫組織化学的研究

    泉健次, 前田健康

    日本口腔外科学会雑誌   38 ( 6 )   896 - 904   1992年6月

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    記述言語:日本語  

    DOI: 10.5794/jjoms.38.896

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  • HISTOCHEMICAL AND IMMUNOHISTOCHEMICAL DEMONSTRATION OF MACROPHAGES AND DENDRITIC CELLS IN THE LINGUAL PERIODONTAL-LIGAMENT OF RAT INCISORS 査読

    KAWAHARA, I, Y TAKANO, O SATO, T MAEDA, K KANNARI

    ARCHIVES OF HISTOLOGY AND CYTOLOGY   55 ( 2 )   211 - 217   1992年5月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:INT SOC HISTOLOGY & CYTOLOGY  

    The distribution of macrophages in the lingual periodontal ligament of rat incisors was surveyed by histochemical and immunohistochemical methods. Numerous macrophages showing intense ACPase reactions were located primarily in the shear zone of the periodontal ligament. Immunostaining with an ED1-monoclonal antibody that recognizes various subpopulations of macrophages revealed plentiful positive cells showing flamelike profiles throughout the periodontal ligament, in addition to regular macrophages associated with sinusoidal blood vessels. A similar distribution of flamelike cells expressing la antigens was demonstrable with immunostaining using an OX6-monoclonal antibody. A consecutive staining of sections for ACPase histochemistry followed by immunoreactions for Ia antigens revealed the presence of two types of the flamelike cells in the periodontal ligament: one with and the other without distinct ACPase activity, corresponding to the macrophage and the dendritic cell, respectively. Either type of flamelike cells was located in the bone-related and shear zones, whereas only dendritic cells without ACPase activity were restricted to the tooth-related zone. OX6-immunonegative cells showing ACPase reactions were also found in the periphery of the sinusoidal blood vessels.
    Our data are the first to demonstrate the abundance of macrophages and dendritic cells expressing Ia antigens throughout the lingual periodontal ligament of rat incisors. In addition to regular macrophages, an exclusive localization of macrophages with flamelike extensions has been demonstrated in the bone-related and shear zones of the ligament. The region-specific arrangement of macrophages and dendritic cells with various histochemical and immunological features suggests that the periodontal ligament of rat incisor is a useful model for analyzing the process of differentiation of antigen-presenting cells.

    DOI: 10.1679/aohc.55.211

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  • INNERVATION OF THE PERIODONTAL-LIGAMENT IN THE DOG WITH SPECIAL REFERENCE TO THE MORPHOLOGY OF RUFFINI ENDINGS 査読

    O SATO, T MAEDA, K KANNARI, KAWAHARA, I, T IWANAGA, Y TAKANO

    ARCHIVES OF HISTOLOGY AND CYTOLOGY   55 ( 1 )   21 - 30   1992年3月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:INT SOC HISTOLOGY & CYTOLOGY  

    The distribution and terminal formation of nerves in the periodontal ligament of dog incisors and canines were investigated by immunohistochemistry for neurofilament protein (NFP) and by electron microscopy. The NFP-immunoreactive nerve fibers were found to be densely distributed in the apical third of the periodontal ligament, while they were sparse in the coronal two thirds. Most of the nerve endings in the periodontal ligament showed a tree-like appearance and resembled those nerve endings demonstrated in the periodontal ligament of human and monkey under the category of free nerve endings. Presumable axon terminals of these were slightly thicker than preterminal portions, running along periodontal collagen fibers and tapering within them. In light microscopic images, at least, they differed from the Ruffini endings which are commonly seen in rodents, displaying a glove-like configuration with extremely expanded tips. Under the electron microscope, however, the tree-like endings of the dog appeared similar to the Ruffini endings of rodents: their terminals were filled with mitochondrial covered with a cytoplasmic process of a Schwann cell, and surrounded by collagen fibers. These ultrastructural findings, combined with the results of previous physiological studies suggest that the nerve endings demonstrated in the present study can be identified as Ruffini endings. It is even stressed that the dog-type of Ruffini ending can be regarded. as a representative of the sensory receptors in the mammalian periodontal ligament. In addition to these endings, knobbed endings, corpuscular (lamellated and glomerular) endings, and free nerve endings were rarely encountered in the periodontal ligament of incisors and canines of the dog.

    DOI: 10.1679/aohc.55.21

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  • Immunocytochemical localization of nerve growth factor receptor (NGFR) in human teeth. 査読

    Maeda T, Sato O, Iwanaga T, Takano Y

    Proceedings of the Finnish Dental Society. Suomen Hammaslaakariseuran toimituksia   88 Suppl 1   557 - 562   1992年

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  • Nerve terminals immunoreactive to neurofilament protein (NFP) in the periodontal ligament of rat molars. 査読

    N. Kamasaki, T. Maeda, K. Izumi, Y. Kozawa

    Jap. J Oral Biol.   34 ( 2 )   239 - 243   1992年

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    記述言語:英語   掲載種別:研究論文(学術雑誌)  

    DOI: 10.2330/joralbiosci1965.34.239

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  • 実験的ハムスター舌癌組織における神経線維の動態:抗NFP、S-100蛋白抗体を用いた免疫組織学的研究 査読

    泉 健次, 前田健康

    口外誌   38 ( 6 )   896 - 904   1992年

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  • A POSSIBLE MECHANISM OF MECHANORECEPTION IN RUFFINI ENDINGS IN THE PERIODONTAL-LIGAMENT OF HAMSTER INCISORS 査読

    K KANNARI, O SATO, T MAEDA, T IWANAGA, T FUJITA

    JOURNAL OF COMPARATIVE NEUROLOGY   313 ( 2 )   368 - 376   1991年11月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:WILEY-LISS  

    The topographical relationship between Ruffini endings and the surrounding collagen fibers in the periodontal ligament of hamster incisors was investigated by means of both immunohistochemistry for neurofilament protein (NFP) and electron microscopy. Periodontal Ruffini endings, a type of stretch receptor, were present exclusively in the alveolar half of the periodontal ligament. Their axon terminals were densely and regularly associated with transverse collagen fibers, possibly forming a mechanoreceptive complex. Since blood sinuses with frequent anastomoses extended throughout the alveolus-related part, the densely innervated collagen bundles were separated from each other by the vascular spaces. Electron microscopic observation of specimens stained with tannic acid revealed a linkage between the axon terminals of the Ruffini endings and the surrounding collagen filaments. The axon terminals were enveloped by multiple layers of the basal lamina, which were penetrated by collagen filaments. The irregularly arranged collagen filaments were sandwiched between electron-dense laminae of the multilayered basal lamina. The possible mechanism of mechanoreception by the periodontal Ruffini endings is discussed on the basis of the immunohistochemical and ultrastructural findings.

    DOI: 10.1002/cne.903130211

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  • IMMUNOHISTOCHEMICAL LOCALIZATION OF LAMININ IN THE PERIODONTAL RUFFINI ENDINGS OF RAT INCISORS - A POSSIBLE FUNCTION OF TERMINAL SCHWANN-CELLS 査読

    T MAEDA, O SATO, K KANNARI, H TAKAGI, T IWANAGA

    ARCHIVES OF HISTOLOGY AND CYTOLOGY   54 ( 3 )   339 - 348   1991年7月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:INT SOC HISTOLOGY & CYTOLOGY  

    Ruffini endings in the periodontal ligament of rodents are ensheathed by a special type of terminal Schwann cell with a particularly developed rough endoplasmic reticulum and Golgi apparatus, and further enveloped by a characteristic multi-layered structure. In order to reveal the functional significance of the structures, localization of a laminin molecule in the periodontal Ruffini endings of rats was immunohistochemically investigated at the levels of light and electron microscopy.
    Immunostaining using an anti-laminin serum clearly demonstrated the profiles of the Ruffini endings as well as those of the blood vessels. Ultrastructurally, reaction products for laminin were deposited in the entire thickness of the multi-layered structure, supporting the idea that this structure is derived from the basal lamina. The basal lamina, immunoreacting with laminin antiserum, was penetrated by periodontal collagen fibers, possibly serving as an adhesive device between the Ruffini endings and surrounding collagen fibers. The laminin immunoreactive materials were also recognized in the vesicles and caveolae of the terminal Schwann cells which tended to gather at the interstitial surface of the cells. The terminal Schwann cells are therefore believed to be directly involved in the formation of the multilayered basal lamina through the active production of its materials.

    DOI: 10.1679/aohc.54.339

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  • Structure and expression of rat S-100 beta subunit gene. 査読

    Maeda T, Usui H, Araki K, Kuwano R, Takahashi Y, Suzuki Y

    Brain research. Molecular brain research   10 ( 3 )   193 - 202   1991年6月

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  • RESPONSES OF CALCITONIN GENE-RELATED PEPTIDE-IMMUNOPOSITIVE NERVE-FIBERS IN THE PERIODONTAL-LIGAMENT OF RAT MOLARS TO EXPERIMENTAL TOOTH MOVEMENT 査読

    SAITO, I, K ISHII, K HANADA, O SATO, T MAEDA

    ARCHIVES OF ORAL BIOLOGY   36 ( 9 )   689 - 692   1991年

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    記述言語:英語   出版者・発行元:PERGAMON-ELSEVIER SCIENCE LTD  

    Nerve fibres showing CGRP-immunoreactivity were exclusively distributed around blood vessels in the periodontal ligament of normal teeth. The CGRP-positive nerves as well as the vessels were more dense during tooth movement and reached a maximum density and intensity of immunoreaction after 3 days. After that, the immunoreactive nerves gradually disappeared, and returned to the control level by the 7th day. Some of the CGRP-nerves were adjacent to bone resorptive lacunae at 3 days when active remodelling of alveolar bone was taking place. These findings suggest that the distribution of CGRP-positive nerves is dynamically changed around the blood vessels in the periodontal ligament during tooth movement, and that CGRP might affect the regulation of blood flow in the ligament. CGRP is possibly involved in the tissue remodelling, directly or indirectly, in addition to its function as a neurotransmitter.

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  • CHOLINESTERASE ACTIVITY IN TERMINAL SCHWANN-CELLS ASSOCIATED WITH RUFFINI ENDINGS IN THE PERIODONTAL-LIGAMENT OF RAT INCISORS 査読

    T MAEDA, K KANNARI, O SATO, S KOBAYASHI, T IWANAGA, T FUJITA

    ANATOMICAL RECORD   228 ( 3 )   339 - 344   1990年11月

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    記述言語:英語   掲載種別:研究論文(学術雑誌)   出版者・発行元:WILEY-LISS  

    DOI: 10.1002/ar.1092280313

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  • [An immunohistochemical study on the response of nerve fibers in the periodontium of rat molars during experimental tooth movement]. 査読

    Saitoh I, Ishii K, Kobayashi M, Hanada K, Maeda T, Sato O

    Nihon Kyosei Shika Gakkai zasshi = The journal of Japan Orthodontic Society   49 ( 5 )   466 - 474   1990年10月

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    記述言語:日本語  

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