Updated on 2024/05/26

写真a

 
OHKURA Naoto
 
Organization
University Medical and Dental Hospital Preventive and Conservative Dentistry Dental Health Assistant Professor
Title
Assistant Professor
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Degree

  • 博士(歯学) ( 2014.3   新潟大学 )

  • 歯学 ( 2014.3   新潟大学 )

  • 薬学修士 ( 2002.3   東北大学 )

Research Interests

  • Dental pulp regeneration

  • 口腔-脳機能連関

  • 歯内療法学

  • 薬物動態学

  • 口腔-胎盤機能連関

Research Areas

  • Life Science / Conservative dentistry

  • Life Science / Clinical pharmacy

  • Life Science / Pharmaceutical chemistry and drug development sciences

Research History (researchmap)

  • University of Michigan, School of Dentistry   Department of Orthodontics and Pediatric Dentistry   Research fellow

    2020.3 - 2022.3

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    Country:United States

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  • Niigata University   Preventive and Conservative Dentistry, Medical and Dental Hospital

    2016.9

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  • Niigata University   Preventive and Conservative Dentistry, Medical and Dental Hospital

    2012.4 - 2016.8

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  • 新潟県 長岡市 私立 立川綜合病院 歯科口腔外科 医員

    2011.4 - 2012.3

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Research History

  • Niigata University   University Medical and Dental Hospital Dental Health Dental Health   Assistant Professor

    2016.9

Education

  • Niigata University   Graduate School of Medical and Dental Sciences   Oral Life Science

    2011.4 - 2014.3

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  • Niigata University   Faculty of Dentistry   School of Dentistry

    2004.4 - 2008.3

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  • 東北大学大学院薬学部博士前期2年の課程 薬物送達学分野

    2002.4 - 2004.3

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  • 富山医科薬科大学 薬学部 薬科学科

    1998.4 - 2002.3

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Professional Memberships

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Qualification acquired

  • Dentist

  • 日本歯内療法学会認定 歯内療法専門医

  • 日本歯科保存学会認定 歯科保存治療認定医

  • Pharmacist

 

Papers

  • Wound-healing Processes After Pulpotomy in the Pulp Tissue of Type 1 Diabetes Mellitus Model Rats

    Rosa Baldeon-Gutierrez, Naoto Ohkura, Kunihiko Yoshiba, Nagako Yoshiba, Aiko Tohma, Ryosuke Takeuchi, Razi Saifullah Ibn Belal, Naoki Edanami, Shintaro Takahara, Susan Gomez-Kasimoto, Takako Ida, Yuichiro Noiri

    Journal of Endodontics   2023.11

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    Authorship:Corresponding author   Publishing type:Research paper (scientific journal)   Publisher:Elsevier BV  

    DOI: 10.1016/j.joen.2023.10.016

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  • Cranial Neural Crest Specific Deletion of Alpl (TNAP) via P0-Cre Causes Abnormal Chondrocyte Maturation and Deficient Cranial Base Growth

    Naoto Ohkura, Hwa Kyung Nam, Fei Liu, Nan Hatch

    International Journal of Molecular Sciences   24 ( 20 )   15401 - 15401   2023.10

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    Authorship:Lead author   Publishing type:Research paper (scientific journal)   Publisher:MDPI AG  

    Bone growth plate abnormalities and skull shape defects are seen in hypophosphatasia, a heritable disorder in humans that occurs due to the deficiency of tissue nonspecific alkaline phosphatase (TNAP, Alpl) enzyme activity. The abnormal development of the cranial base growth plates (synchondroses) and abnormal skull shapes have also been demonstrated in global Alpl−/− mice. To distinguish local vs. systemic effects of TNAP on skull development, we utilized P0-Cre to knockout Alpl only in cranial neural crest-derived tissues using Alpl flox mice. Here, we show that Alpl deficiency using P0-Cre in cranial neural crest leads to skull shape defects and the deficient growth of the intersphenoid synchondrosis (ISS). ISS chondrocyte abnormalities included increased proliferation in resting and proliferative zones with decreased apoptosis in hypertrophic zones. ColX expression was increased, which is indicative of premature differentiation in the absence of Alpl. Sox9 expression was increased in both the resting and prehypertrophic zones of mutant mice. The expression of Parathyroid hormone related protein (PTHrP) and Indian hedgehog homolog (IHH) were also increased. Finally, cranial base organ culture revealed that inorganic phosphate (Pi) and pyrophosphate (PPi) have specific effects on cell signaling and phenotype changes in the ISS. Together, these results demonstrate that the TNAP expression downstream of Alpl in growth plate chondrocytes is essential for normal development, and that the mechanism likely involves Sox9, PTHrP, IHH and PPi.

    DOI: 10.3390/ijms242015401

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  • Prostaglandin E2-Transporting Pathway and Its Roles via EP2/EP4 in Cultured Human Dental Pulp

    Naoto Ohkura, Kunihiko Yoshiba, Nagako Yoshiba, Yohei Oda, Naoki Edanami, Hayato Ohshima, Shoji Takenaka, Takashi Okiji, Yuichiro Noiri

    Journal of Endodontics   49 ( 4 )   410 - 418   2023.4

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    Authorship:Lead author, Corresponding author   Publishing type:Research paper (scientific journal)   Publisher:Elsevier BV  

    DOI: 10.1016/j.joen.2023.01.009

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  • SVCT2-GLUT1-mediated ascorbic acid transport pathway in rat dental pulp and its effects during wound healing

    Naoto Ohkura, Kunihiko Yoshiba, Nagako Yoshiba, Naoki Edanami, Hayato Ohshima, Shoji Takenaka, Yuichiro Noiri

    Scientific Reports   13 ( 1 )   2023.1

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    Authorship:Lead author, Corresponding author   Publishing type:Research paper (scientific journal)   Publisher:Springer Science and Business Media LLC  

    Abstract

    Ascorbic acid (AA; vitamin C) plays a crucial role in the biosynthesis and secretion of collagen to produce the organic matrix of hard tissues. Nevertheless, the detailed mechanism by which AA induces reparative dentinogenesis is still unknown. This study aimed to investigate the pathway and function of AA during wound healing in a rat pulpotomy model. Sodium-dependent vitamin C transporter (SVCT) 2 and glucose transporter (GLUT) 1 were detected in odontoblasts, endothelial cells, and nerve fibers in normal pulp tissues. SVCT2 and GLUT1 were also expressed in odontoblast-like cells in pulpotomized tissues of Wistar rats, and immunopositive cells of SVCT2 were significantly increased at 5 days after pulpotomy (p < 0.05). By contrast, osteogenic disorder Shionogi (ODS) rats, which cannot generate AA, also expressed SVCT2 and GLUT1 in normal and wound healing conditions. However, in ODS rats, when compared with the AA-addition group, the formation of dentin bridges in the AA-loss group was not evident, a layer of osteopontin was significantly increased beneath the wound surface (p < 0.05), and alpha smooth muscle actin at the odontoblast-like cells observed along this layer was significantly increased (p < 0.05), but not Nestin. Moreover, the amounts of type 1 collagen generated in the reparative dentin and beneath the wound healing site were significantly diminished (p < 0.05). Macrophages expressing CD68 and CD206 increased beneath the wound site. Hence, AA may be involved in odontoblast-like cell differentiation and anti-inflammatory response during dental pulp wound healing. Our results provide new insights into the function of AA through SVCT2 and GLUT1 in reparative dentinogenesis and may help in developing new therapeutic targets for dental pulpal disease.

    DOI: 10.1038/s41598-023-28197-9

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    Other Link: https://www.nature.com/articles/s41598-023-28197-9

  • Impact of remnant healthy pulp and apical tissue on outcomes after simulated regenerative endodontic procedure in rat molars. International journal

    Naoki Edanami, Kunihiko Yoshiba, Mari Shirakashi, Razi Saifullah Ibn Belal, Nagako Yoshiba, Naoto Ohkura, Aiko Tohma, Ryosuke Takeuchi, Takashi Okiji, Yuichiro Noiri

    Scientific reports   10 ( 1 )   20967 - 20967   2020.12

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    When regenerative endodontic procedures (REPs) are performed on immature teeth diagnosed with pulp necrosis and apical periodontitis, various healing patterns occur. Furthermore, infected immature teeth with endodontic disorders often exhibit some remnant pulp and apical tissue. Therefore, this study investigated the impact of remnant healthy or fully functional pulp and apical tissue on healing patterns after REPs. Simulated REPs were performed on non-infected immature rat molars with different amounts of remnant pulp and apical tissue. Healing patterns in these teeth were assessed after 28 days. Teeth with 0.81-0.91 mm of remnant pulp healed with pulp-like tissue, dentin, and osteodentin-like dentin-associated mineralized tissue (OSD-DAMT); teeth with 0.60-0.63 mm of remnant pulp healed with pulp-like tissue and OSD-DAMT; teeth with 0.13-0.43 mm of remnant pulp healed with periodontal ligament (PDL)-like tissue, OSD-DAMT, and cementum-like dentin-associated mineralized tissue (CEM-DAMT); and teeth with disorganization of pulp and apical tissues at 0.15-0.38 mm beyond the root apex healed with PDL-like tissue, CEM-DAMT, and intracanal bone (IB). Loss of Hertwig's epithelial root sheath was observed with IB formation. These results showed that four distinct healing patterns occurred after REPs, depending on the preoperative amount of remnant healthy pulp and apical tissue.

    DOI: 10.1038/s41598-020-78022-w

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  • Glucose Transporter 2 and 4 Are Involved in Glucose Supply during Pulpal Wound Healing after Pulpotomy with Mineral Trioxide Aggregate in Rat Molars

    Aiko Tohma, Naoto Ohkura, Kunihiko Yoshiba, Ryosuke Takeuchi, Nagako Yoshiba, Naoki Edanami, Mari Shirakashi, Razi Saifullah Ibn Belal, Hayato Ohshima, Yuichiro Noiri

    Journal of Endodontics   46 ( 1 )   81 - 88   2020.1

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    Authorship:Corresponding author   Publishing type:Research paper (scientific journal)   Publisher:Elsevier BV  

    DOI: 10.1016/j.joen.2019.10.003

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  • Immunohistochemistry and gene expression of GLUT1, RUNX2 and MTOR in reparative dentinogenesis

    Ryosuke Takeuchi, Naoto Ohkura, Kunihiko Yoshiba, Aiko Tohma, Nagako Yoshiba, Naoki Edanami, Mari Shirakashi, Razi Saifullah Ibn Belal, Hayato Ohshima, Yuichiro Noiri

    Oral Diseases   26 ( 2 )   341 - 349   2019.12

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    Authorship:Corresponding author   Publishing type:Research paper (scientific journal)   Publisher:Wiley  

    Abstract

    Objectives

    To determine glucose transporter 1 (GLUT1) and runt‐related transcription factor 2 (RUNX2) expression during reparative dentinogenesis after pulpotomy with mineral trioxide aggregate (MTA) capping.

    Subjects and methods

    Eight‐week‐old male Wistar rats were used. Pulp of the upper left first molar was exposed and capped with MTA. The upper right first molar of the same animal was used as a control. After collecting molars at various time points, GLUT1, RUNX2 and mammalian target of rapamycin (MTOR) were examined by immunohistochemistry. mRNA levels of Slc2a1 (encoding GLUT1), Runx2, Nestin and Mtor were determined by real‐time PCR.

    Results

    Pulp exhibited progressive formation of reparative dentine lined with GLUT1‐ and MTOR‐immunoreactive odontoblast‐like cells at 5 days after pulpotomy. RUNX2 was detected in nuclei of most pulp tissue cells at day 5 after pulpotomy. Double immunofluorescence staining revealed GLUT1 immunoreactivity on odontoblast‐like cells positive for Nestin or RUNX2, 5 days after pulpotomy. Slc2a1, Runx2, Nestin and Mtor mRNA levels were significantly upregulated on days 3–5 after pulpotomy.

    Conclusions

    After rat molar pulpotomy, dental pulp induced formation of reparative dentine with colocalization of GLUT1 and Nestin or RUNX2. Moreover, mRNA levels of Slc2a1, Runx2, Nestin and Mtor were significantly upregulated in pulpotomized dental pulp.

    DOI: 10.1111/odi.13230

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    Other Link: https://onlinelibrary.wiley.com/doi/full-xml/10.1111/odi.13230

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

    Journal of Dental Research   論文受理 ( 印刷中 )   2019.11

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  • Orthodontic force application upregulated pain-associated prostaglandin-I2/PGI2-receptor/TRPV1 pathway-related gene expression in rat molars. Reviewed

    Mariko Ohkura, Naoto Ohkura, Nagako Yoshiba, Kunihiko Yoshiba, Hiroko Ida-Yonemochi, Hayato Ohshima, Isao Saito, Takashi Okiji

    Odontology   106 ( 1 )   2 - 10   2018.1

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    This study aimed to analyze the mRNA expression and protein localization of prostaglandin I2 (PGI2) synthase (PGIS), the PGI2 receptor (IP receptor) and transient receptor potential cation channel, subfamily V, member 1 (TRPV1) in force-stimulated rat molars, toward the elucidation of the PGI2-IP receptor-TRPV1 pathway that is in operation in the pulp and possibly associated with orthodontic pain and inflammation. Experimental force was applied to the maxillary first and second molars by inserting an elastic band between them for 6-72 h. PGIS, PTGIR (the IP receptor gene), and TRPV1 mRNA levels in the coronal pulp were analyzed with real-time PCR. PGIS, IP receptor, and TRPV1 proteins were immunostained. The force stimulation induced significant upregulation of PGIS at 6-24 h, and PTGIR and TRPV1 at 6 and 12 h in the pulp. PGIS was immunolocalized in odontoblasts and some fibroblasts in the force-stimulated pulp. The IP receptor and TRPV1 immunoreactivities were detected on odontoblasts and some nerve fibers. It was concluded that PGIS, PTGIR, and TRPV1 in rat molar pulp were significantly upregulated shortly after the force application, and that the IP receptor was co-expressed on TRPV1-expressing nerves and odontoblasts. These findings suggest that the PGI2-IP receptor-TRPV1 pathway is associated with the acute phase of force-induced pulp changes involving odontoblasts and nerves.

    DOI: 10.1007/s10266-017-0309-2

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  • Effects of pulpotomy using mineral trioxide aggregate on prostaglandin transporter and receptors in rat molars. Reviewed International journal

    Naoto Ohkura, Naoki Edanami, Ryosuke Takeuchi, Aiko Tohma, Mariko Ohkura, Nagako Yoshiba, Kunihiko Yoshiba, Hiroko Ida-Yonemochi, Hayato Ohshima, Takashi Okiji, Yuichiro Noiri

    Scientific reports   7 ( 1 )   6870 - 6870   2017.7

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    Authorship:Lead author, Corresponding author   Language:English   Publishing type:Research paper (scientific journal)  

    Mineral trioxide aggregate (MTA) is a commonly used dental pulp-capping material with known effects in promoting reparative dentinogenesis. However, the mechanism by which MTA induces dentine repair remains unclear. The aim of the present study was to investigate the role of prostaglandin E2 (PGE2) in dentine repair by examining the localisation and mRNA expression levels of its transporter (Pgt) and two of its receptors (Ep2 and Ep4) in a rat model of pulpotomy with MTA capping. Ep2 expression was detected in odontoblasts, endothelial cells, and nerve fibres in normal and pulpotomised tissues, whereas Pgt and Ep4 were immunolocalised only in the odontoblasts. Moreover, mRNA expression of Slco2a1 (encoding Pgt), Ptger2 (encoding Ep2), and Ptger4 (encoding Ep4) was significantly upregulated in pulpotomised dental pulp and trigeminal ganglia after MTA capping. Our results provide insights into the functions of PGE2 via Pgt and Ep receptors in the healing dentine/pulp complex and may be helpful in developing new therapeutic targets for dental disease.

    DOI: 10.1038/s41598-017-07167-y

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  • Prostaglandin transporting protein-mediated prostaglandin E2 transport in lipopolysaccharide-inflamed rat dental pulp. Reviewed International journal

    Naoto Ohkura, Yoshimi Shigetani, Nagako Yoshiba, Kunihiko Yoshiba, Takashi Okiji

    Journal of endodontics   40 ( 8 )   1112 - 7   2014.8

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    Authorship:Lead author   Language:English   Publishing type:Research paper (scientific journal)   Publisher:ELSEVIER SCIENCE INC  

    INTRODUCTION: The prostaglandin transporter (Pgt) and multidrug resistance-associated protein (Mrp) 4 are membrane transport proteins that play crucial roles in the transmembrane uptake and/or efflux of prostaglandins (PGs). This study attempted to analyze the protein expression of Pgt and Mrp4 and their involvement in PGE2 efflux transport in lipopolysaccharide (LPS)-inflamed rat incisor pulp tissue. METHODS: Pulpitis was induced in the upper incisors of Wistar rats by treating them with LPS for 24 hours. The protein expression levels of Pgt, Mrp4, and microsomal PGE synthase (mPGES) were analyzed with immunofluorescent staining. The amount of PGE2 released from the inflamed pulp tissue in the presence or absence of dipyridamole (an Mrp4 inhibitor) was assessed by using an enzyme-linked immunosorbent assay. RESULTS: Double immunofluorescence staining revealed that the Pgt, Mrp4, and mPGES immunoreactivity co-localized in CD31-expressing endothelial cells. Moreover, the Mrp4 inhibitor caused a significant decrease in the amount of PGE2 released from the LPS-inflamed pulp (P < .01 at 24 hours). CONCLUSION: Pgt, Mrp4, and mPGES expression was detected in the endothelial cells of normal and LPS-inflamed rat incisor pulp tissue, suggesting that these cells are associated with the biosynthesis and transmembrane transport of PGE2. The significant decrease in PGE2 release induced by the Mrp4 inhibitor suggests that Mrp4 contributes to the transport of PGE2 in the transmembrane efflux pathway.

    DOI: 10.1016/j.joen.2013.12.024

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  • Autologous concentrated growth factor mediated accelerated bone healing in root-end microsurgery: A multicenter randomized clinical trial. International journal

    Yoshio Yahata, Keisuke Handa, Naoto Ohkura, Motoki Okamoto, Jun Ohshima, Shusaku Itoh, Nobuyuki Kawashima, Toshinori Tanaka, Nobuya Sato, Yuichiro Noiri, Mikako Hayashi, Takashi Okiji, Masahiro Saito

    Regenerative therapy   24   377 - 384   2023.12

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    INTRODUCTION: Concentrated growth factor (CGF) is a new-generation autologous platelet concentrate that promotes tissue regeneration and has anti-inflammatory properties. This randomized multicenter trial aimed to evaluate the effects of CGF on bone healing in combination with root-end microsurgery. METHODS: Healthy adult patients indicated for root-end microsurgery were randomly assigned to either the CGF or control (no CGF implantation) groups. CGF was implanted into the bone cavity after root-end filling with mineral trioxide aggregate. Clinical and periapical radiographic evaluations were conducted at 1, 3, 6, and 12 months postoperatively, with follow-up cone-beam computed tomography (CBCT) at 6 months. The lesion volume reduction rate was calculated based on data from the preoperative and follow-up CBCT images. RESULTS: A total of 24 patients were enrolled. The treatment success rate was 91.7% and 83.3% on 12-month periapical radiography and 6-month CBCT, respectively, without a significant difference between the two groups. The lesion volume reduction rate in the CGF group (75.6%) was significantly higher than that in the control (61.0%) group. CONCLUSIONS: Autologous CGF in conjunction with root-end microsurgery accelerated lesion reduction as observed on CBCT. Administering autologous blood products to stimulate healing in addition to removing the source of infection appears to be a promising treatment option for root-end microsurgery.

    DOI: 10.1016/j.reth.2023.08.006

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  • Preparing of Point-of-Care Reagents for Risk Assessment in the Elderly at Home by a Home-Visit Nurse and Verification of Their Analytical Accuracy Reviewed

    Shoji Takenaka, Hiroshi Moro, Utako Shimizu, Takeshi Koizumi, Kei Nagano, Naoki Edanami, Naoto Ohkura, Hisanori Domon, Yutaka Terao, Yuichiro Noiri

    Diagnistics   ahead of online ( 14 )   2407 - 2407   2023.7

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    Language:English   Publishing type:Research paper (scientific journal)   Publisher:MDPI AG  

    With the rising number of older adults residing at home, there is a growing need for risk assessment and patient management in home nursing. This study aims to develop point-of-care test (POCT) reagents that can aid in risk assessment and home care, especially in settings with limited resources. Our focus was on creating a C-reactive protein (CRP) POCT, which can accurately diagnose clinically significant judgment values in home nursing. Additionally, we assessed the utility of the HemoCue WBC DIFF system in providing differential counts of white blood cells (WBC). These performances were compared with a laboratory test using blood samples from patients with pneumonia. The CRP POCT showed a comparable result to that of a laboratory method, with an average kappa index of 0.883. The leukocyte count showed good agreement with the reference method. While the correlation coefficients for both neutrophil and lymphocyte counts were deemed acceptable, it was observed that the measured values tended to be smaller in cases where the cell count was higher. This proportional error indicates a weak correlation with the neutrophil-to-lymphocyte ratio. CRP POCT and WBC counts provided reliable and accurate judgments. These tools may benefit risk management for older adults at home, patients with dementia who cannot communicate, and those living in depopulated areas.

    DOI: 10.3390/diagnostics13142407

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  • In Vivo assessment of the Apatite-Forming Ability of New-Generation Hydraulic Calcium Silicate Cements Using a Rat Subcutaneous Implantation Model Reviewed International journal

    Naoki Edanami, Shouji Takenaka, Razi Saifullah, Ibn Belal, Kunihiko Yoshiba, Shintaro Takahara, Nagako Yoshiba, Naoto Ohkura, Yuichiro Noiri

    Journal of Functional Biomaterials   14 ( 213 )   ahead of online   2023.4

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    Hydroxyapatite formation on endodontic hydraulic calcium silicate cements (HCSCs) plays a significant role in sealing the root canal system and elevating the hard-tissue inductivity of the materials. This study evaluated the in vivo apatite-forming ability of 13 new-generation HCSCs using an original HCSC (white ProRoot MTA: PR) as a positive control. The HCSCs were loaded into polytetrafluoroethylene tubes and implanted in the subcutaneous tissue of 4-week-old male Wistar rats. At 28 days after implantation, hydroxyapatite formation on the HCSC implants was assessed with micro-Raman spectroscopy, surface ultrastructural and elemental characterization, and elemental mapping of the material-tissue interface. Seven new-generation HCSCs and PR had a Raman band for hydroxyapatite (v1 PO43- band at 960 cm-1) and hydroxyapatite-like calcium-phosphorus-rich spherical precipitates on the surfaces. The other six HCSCs with neither the hydroxyapatite Raman band nor hydroxyapatite-like spherical precipitates did not show calcium-phosphorus-rich hydroxyapatite-layer-like regions in the elemental mapping. These results indicated that 6 of the 13 new-generation HCSCs possessed little or no ability to produce hydroxyapatite in vivo, unlike PR. The weak in vivo apatite-forming ability of the six HCSCs may have a negative impact on their clinical performance.

    DOI: 10.3390/jfb14040213

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  • In Vivo Assessment of the Calcium Salt-Forming Ability of a New Calcium Silicate-Based Intracanal Medicament: Bio-C Temp. International journal

    Naoki Edanami, Razi Saifullah Ibn Belal, Shoji Takenaka, Kunihiko Yoshiba, Rosa Edith Baldeon Gutierrez, Shintaro Takahara, Nagako Yoshiba, Naoto Ohkura, Yuichiro Noiri

    Dentistry journal   11 ( 4 )   2023.3

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    Calcium salt precipitation induced by intracanal medicaments contributes to the formation of apical hard tissue during apexification. This study compared the calcium salt-forming ability of a new calcium silicate-based intracanal medicament (Bio-C Temp) with that of two commercial calcium hydroxide pastes (Calcipex Plane II and Vitapex) in a rat subcutaneous implantation model. Polytetrafluoroethylene tubes containing each of the three materials were subcutaneously implanted in 4-week-old male Wistar rats. After 28 days, the composition and amount of calcium salts formed at the material-tissue interface were assessed using micro-Raman spectroscopy, X-ray diffraction, and elemental mapping. The tested materials produced white precipitates that had Raman spectra with peaks corresponding to hydroxyapatite and calcite. X-ray diffraction detected hydroxyapatite formation on Calcipex Plane II and Vitapex implants, as well as calcite formation on all three materials. Elemental mapping revealed that Bio-C Temp generated significantly smaller calcium- and phosphorus-rich calcified regions within the subcutaneous connective tissue than Vitapex. These results indicate that Bio-C Temp produced less calcium salt in rat subcutaneous tissue than Vitapex, although all materials formed hydroxyapatite and calcite in rat subcutaneous tissue. Bio-C Temp could be less effective than Vitapex in promoting apical hard tissue formation during apexification.

    DOI: 10.3390/dj11040091

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  • Effect of a resin-modified calcium silicate cement on inflammatory cell infiltration and reparative dentin formation after pulpotomy in rat molars. International journal

    Naoki Edanami, Razi Saifullah Ibn Belal, Kunihiko Yoshiba, Nagako Yoshiba, Naoto Ohkura, Shoji Takenaka, Yuichiro Noiri

    Australian endodontic journal : the journal of the Australian Society of Endodontology Inc   48 ( 2 )   297 - 304   2022.8

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    Resin monomers and polymerisation initiators have been shown to be cytotoxic for pulp cells and to disturb odontoblast differentiation. This study aimed to compare the effect of a resin-modified calcium silicate cement (TheraCal LC; TC) and a resin-free calcium silicate cement (ProRoot MTA; PR) on pulpal healing after pulpotomy. Pulpotomy was performed on the maxillary first molars of 8-week-old rats using either PR or TC. After 1, 3, 7, 14 and 28 days, pulpal responses were assessed by micro-computed tomography, haematoxylin-eosin staining and immunostaining against CD68, which is a pan-macrophage marker. The results showed that pulpotomy with TC induced persistent infiltration of inflammatory cells, including CD68-positive macrophages, and delayed the formation of reparative dentin as compared with that with PR, although both materials allowed pulpal healing over the long term. Therefore, resin-modified TC was not as biocompatible nor bioinductive as resin-free PR when applied on the healthy pulp of rat molars.

    DOI: 10.1111/aej.12568

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  • Impact of the pyrophosphate and inorganic phosphate during the root formation

    大倉直人, 吉羽邦彦, 枝並直樹, 竹中彰治, 野杁由一郎

    日本歯内療法学会学術大会プログラム・抄録集   43回   67 - 67   2022.7

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  • Evidence on the Use of Mouthwash for the Control of Supragingival Biofilm and Its Potential Adverse Effects. Reviewed International journal

    Shoji Takenaka, Maki Sotozono, Naoto Ohkura, Yuichiro Noiri

    Antibiotics (Basel, Switzerland)   11 ( 6 )   2022.5

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    Antimicrobial mouthwash improves supragingival biofilm control when used in conjunction with mechanical removal as part of an oral hygiene routine. Mouthwash is intended to suppress bacterial adhesion during biofilm formation processes and is not aimed at mature biofilms. The most common evidence-based effects of mouthwash on the subgingival biofilm include the inhibition of biofilm accumulation and its anti-gingivitis property, followed by its cariostatic activities. There has been no significant change in the strength of the evidence over the last decade. A strategy for biofilm control that relies on the elimination of bacteria may cause a variety of side effects. The exposure of mature oral biofilms to mouthwash is associated with several possible adverse reactions, such as the emergence of resistant strains, the effects of the residual structure, enhanced pathogenicity following retarded penetration, and ecological changes to the microbiota. These concerns require further elucidation. This review aims to reconfirm the intended effects of mouthwash on oral biofilm control by summarizing systematic reviews from the last decade and to discuss the limitations of mouthwash and potential adverse reactions to its use. In the future, the strategy for oral biofilm control may shift to reducing the biofilm by detaching it or modulating its quality, rather than eliminating it, to preserve the benefits of the normal resident oral microflora.

    DOI: 10.3390/antibiotics11060727

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  • Is bioactivity a universal property of calcium silicate cements?-In vivo evaluation of 18 calcium silicate cements-

    枝並直樹, IBN BELAL Razi Saifullah, 竹中彰治, 吉羽邦彦, 大倉直人, 吉羽永子, 野杁由一郎

    日本歯科保存学会学術大会プログラムおよび講演抄録集(Web)   156回   24 - 24   2022.5

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  • Effects of rice fermented extracts, "Sake Lees", on the functional activity of odontoblast-like cells (KN-3 cells).

    Keiichiro Okamoto, Yoshito Kakihara, Naoto Ohkura, Aiko Tohma, Ayako Washio, Chiaki Kitamura, Yuichiro Noiri, Kensuke Yamamura, Makio Saeki

    Odontology   110 ( 2 )   254 - 261   2022.4

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    This study was designed to investigate the effects of Sake Lees extracts (SLE, Sake Kasu) on the functional activity of odontoblastic cells and tooth pulp of the rats. For in vitro studies, a rat clonal odontoblast-like cell line, KN-3 cells were cultured. SLE significantly decreased KN-3 cell proliferation, but showed no significant cytotoxicity. SLE effects on several protein productions of KN-3 cells were compared with PBS. SLE and PBS increased alkaline phosphatase (ALP), dentin sialoprotein (DSP), and osterix in a day-course dependent manner, while SLE increased the induction of ALP on day 9-21 and DSP on day 15-21. SLE also increased Runx2 expression on day 3 and 9 compared to PBS. Alizarin Red stainings revealed that SLE showed a subtle increase in mineralization of KN-3 cells on day 15 and 21. A histological investigation was conducted to assess if SLE induced reparative dentin formation after direct capping at the exposed tooth pulp in rats, suggesting that SLE could increase the reparative dentin formation more than PBS. These findings suggest that Sake Lees could have functional roles in the alterations of odontoblastic activity, which might influence the physiology of the tooth pulp.

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  • Comparison of calcium and hydroxyl ion release ability and in vivo apatite-forming ability of three bioceramic-containing root canal sealers. International journal

    Razi Saifullah Ibn Belal, Naoki Edanami, Kunihiko Yoshiba, Nagako Yoshiba, Naoto Ohkura, Shoji Takenaka, Yuichiro Noiri

    Clinical oral investigations   26 ( 2 )   1443 - 1451   2022.2

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    OBJECTIVE: Bioceramic-containing root canal sealers promote periapical healing via Ca2+ and OH- release and apatite formation on the surface. This study aimed to compare Ca2+ and OH- release and in vivo apatite formation of three bioceramic-containing root canal sealers: EndoSequence BC sealer (Endo-BC), MTA Fillapex (MTA-F), and Nishika Canal Sealer BG (N-BG). MATERIALS AND METHODS: Polytetrafluoroethylene tubes filled with sealers were immersed in distilled water for 6 and 12 h and for 1, 7, 14, and 28 days to measure Ca2+ and OH- release. Additionally, tubes filled with sealers were implanted in the backs of rats for 28 days, and in vivo apatite formation was analyzed using an electron probe microanalyzer. RESULTS: Endo-BC released significantly more Ca2+ than the other sealers at 6 and 12 h and 1 day. Ca2+ release was significantly lower from N-BG than from Endo-BC and MTA-F at 14 and 28 days. OH- release was significantly higher from Endo-BC than from the other sealers throughout the experiment, except at 1 day. OH- release was lower from N-BG than from MTA-F at 6 h and 7 days. Only Endo-BC implants exhibited apatite-like calcium-, phosphorus-, oxygen-, and carbon-rich spherulites and apatite layer-like calcium- and phosphorus-rich, but radiopaque element-free, surface regions. CONCLUSIONS: Ca2+ and OH- release is ranked as follows: Endo-BC > MTA-F > N-BG. Only Endo-BC demonstrated in vivo apatite formation. CLINICAL RELEVANCE: Endo-BC could promote faster periapical healing than MTA-F and N-BG.

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  • Laminin Isoforms in Human Dental Pulp: Lymphatic Vessels Express Laminin-332, and Schwann Cell-Associated Laminin-211 Modulates CD163 Expression of M2-like Macrophages. International journal

    Nagako Yoshiba, Naoki Edanami, Naoto Ohkura, Tomoki Maekawa, Naoki Takahashi, Takahiro Tsuzuno, Takeyasu Maeda, Koichi Tabeta, Kenji Izumi, Yuichiro Noiri, Kunihiko Yoshiba

    ImmunoHorizons   5 ( 12 )   1008 - 1020   2021.12

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    Laminin, a basement membrane heterotrimeric glycoprotein composed of α/β/γ subunits, has important tissue-specific functions in the control of cellular behavior. Our recent study showed the colocalization of CD163+ M2-like macrophages with Schwann cells in human dental pulp, leading us to hypothesize that the laminin isoform of Schwann cells is associated with CD163 expression. The present study investigated the distribution of laminin isoforms in human dental pulp and the underlying mechanisms that affect macrophage phenotypes. Immunofluorescence analysis indicated that blood vessels were exclusively positive for laminin α4 and α5, whereas laminin α2 was associated with Schwann cells. Unexpectedly, laminin α3/laminin-332 (α3β3γ2) was detected on lymphatic vessels. In intact and carious teeth, CD163+ cells were associated with laminin α2, whereas CD206 single-positive cells were present inside, outside, and along blood vessels. In vitro incubation of THP-1 macrophages in plates coated with laminin-211/511 or its functionally analogous E8 fragments of α-chain (E8-α) indicated that cell shapes differed between macrophages grown on laminin-211/E8-α2 and macrophages grown on laminin-511/E8-α5. Laminin-211/E8-α2-coated plates upregulated CD163 expression, compared with laminin-511/E8-α5-coated plates. Integrin α3- and integrin α6-neutralizing Abs altered the shape of THP-1 macrophages and upregulated mRNA levels of CD206 and CD163 in macrophages grown on laminin-511; the neutralizing Abs did not affect macrophages grown on laminin-211. These findings suggest that laminin isoforms differentially regulate macrophage behavior via distinct integrin-laminin affinities. Of note, laminin-332 is expressed by pulpal lymphatic vessels, the existence of which has been debated; laminin-211 might have a role in maintaining CD163 expression on macrophages.

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  • Apatite-Forming Ability of Flowable vs. Putty Formulations of Newly Developed Bioactive Glass-Containing Endodontic Cement

    Naoki Edanami, Razi Saifullah Ibn Belal, Shoji Takenaka, Kunihiko Yoshiba, Nagako Yoshiba, Naoto Ohkura, Shintaro Takahara, Yuichiro Noiri

    Applied Sciences   11 ( 19 )   8969 - 8969   2021.9

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    This study compared the apatite-forming ability (AFA) levels of flowable and putty formulations of Nishika Canal Sealer BG Multi (F-NBG and P-NBG, respectively) and attempted to clarify the cause of differences in the AFA levels of F-NBG and P-NBG. NBG samples were aged in simulated body fluid (SBF) or 1-, 5-, or 10-g/L bovine serum albumin-containing SBF (BSA-SBF) and analyzed in terms of their ultrastructures, elemental compositions, and Raman spectra to identify apatite formation. The phosphate ion consumption rates of NBG samples in the media were evaluated as an indicator of apatite growth. The original elemental composition, calcium ion release, and alkalizing ability levels of F-NBG and P-NBG were also evaluated. Apparent apatite formation was detected on all NBG samples except F-NBG aged in 10-g/L BSA-SBF. P-NBG consumed phosphate ions faster than F-NBG. As-prepared P-NBG showed more silicon elements on its surface than as-prepared F-NBG. P-NBG released more calcium ions than F-NBG, although their alkalizing ability levels did not differ statistically. In conclusion, the AFA of P-NBG was greater than that of F-NBG, probably because of the greater ability of P-NBG to expose silanol groups on the surface and release calcium ions.

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  • 各種バイオセラミック系覆髄材のアパタイト析出能に関する研究

    枝並 直樹, イブンベラル・ラジサイフラー, 白柏 麻里, 吉羽 邦彦, 大倉 直人, 吉羽 永子, 遠間 愛子, 竹内 亮祐, 野杁 由一郎

    特定非営利活動法人日本歯科保存学会学術大会プログラムおよび講演抄録集   152回   97 - 97   2020.6

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  • in vitro・in vivoにおける各種バイオセラミック系シーラーのアパタイト析出能に関する研究(Apatite forming ability of different bioceramic based root canal sealers in vitro and in vivo)

    イブンベラル・ラジサイフラー, 枝並 直樹, 白柏 麻里, 吉羽 邦彦, 大倉 直人, 吉羽 永子, 遠間 愛子, 竹内 亮祐, 野杁 由一郎

    特定非営利活動法人日本歯科保存学会学術大会プログラムおよび講演抄録集   152回   46 - 46   2020.6

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  • Comparison of apatite forming ability of various bio-ceramic based pulp capping materials

    枝並直樹, SAIFULLAH Ibn Belal Razi, 白柏麻里, 吉羽邦彦, 大倉直人, 吉羽永子, 遠間愛子, 竹内亮祐, 野杁由一郎

    日本歯科保存学会学術大会プログラムおよび講演抄録集(Web)   152回   97 - 97   2020.6

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  • in vitro・in vivoにおける各種バイオセラミック系シーラーのアパタイト析出能に関する研究(Apatite forming ability of different bioceramic based root canal sealers in vitro and in vivo)

    イブンベラル・ラジサイフラー, 枝並 直樹, 白柏 麻里, 吉羽 邦彦, 大倉 直人, 吉羽 永子, 遠間 愛子, 竹内 亮祐, 野杁 由一郎

    特定非営利活動法人日本歯科保存学会学術大会プログラムおよび講演抄録集   152回   46 - 46   2020.6

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  • Anti-biofilm Property of Sulfated Vizantin Against Streptococcus mutans -Influence of Sucrose Included in Growth Media and Expression Analysis of Genes Related to Biofilm Formation- Reviewed

    竹中彰治, 長谷川泰輔, 小田真隆, 高橋直紀, 磯野俊仁, 大倉直人, 山本博文, 多部田康一, 野杁由一郎

    日本歯科保存学雑誌   63 ( 1 )   61 - 72   2020.2

  • なぜRegenerative endodontic proceduresは多様な治癒形態を生じさせるのか ラット根未完成臼歯における免疫組織学的研究 Reviewed

    枝並 直樹, 白柏 麻里, 吉羽 邦彦, 大倉 直人, 吉羽 永子, 遠間 愛子, 竹内 亮祐, 野杁 由一郎

    特定非営利活動法人日本歯科保存学会学術大会プログラムおよび講演抄録集   151回   54 - 54   2019.10

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  • 糖尿病モデルラットを用いた修復象牙質形成時におけるマクロファージの挙動と細胞増殖の解析 Reviewed

    遠間 愛子, 大倉 直人, 吉羽 邦彦, 吉羽 永子, 枝並 直樹, 白柏 麻里, 竹内 亮祐, 野杁 由一郎

    特定非営利活動法人日本歯科保存学会学術大会プログラムおよび講演抄録集   151回   109 - 109   2019.10

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  • なぜRegenerative endodontic proceduresは多様な治癒形態を生じさせるのか ラット根未完成臼歯における免疫組織学的研究

    枝並 直樹, 白柏 麻里, 吉羽 邦彦, 大倉 直人, 吉羽 永子, 遠間 愛子, 竹内 亮祐, 野杁 由一郎

    特定非営利活動法人日本歯科保存学会学術大会プログラムおよび講演抄録集   151回   54 - 54   2019.10

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  • 糖尿病モデルラットを用いた修復象牙質形成時におけるマクロファージの挙動と細胞増殖の解析

    遠間 愛子, 大倉 直人, 吉羽 邦彦, 吉羽 永子, 枝並 直樹, 白柏 麻里, 竹内 亮祐, 野杁 由一郎

    特定非営利活動法人日本歯科保存学会学術大会プログラムおよび講演抄録集   151回   109 - 109   2019.10

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  • ヒト歯髄においてシュワン細胞はマクロファージをM2型へ転換する

    吉羽 永子, 大倉 直人, 前川 知樹, 泉 健次, 細矢 明宏, 中村 浩彰, 前田 健康, 野杁 由一郎, 吉羽 邦彦, 枝並 直樹, 遠間 愛子

    Journal of Oral Biosciences Supplement   2019   302 - 302   2019.10

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  • ラット臼歯歯髄における創傷治癒時のグルタミン輸送担体の解析 Reviewed

    大倉 直人, 吉羽 邦彦, 吉羽 永子, 枝並 直樹, 遠間 愛子, 竹内 亮祐, 白柏 麻里, 野杁 由一郎

    特定非営利活動法人日本歯科保存学会学術大会プログラムおよび講演抄録集   150回   40 - 40   2019.5

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  • Detection of bone marrow-derived fibrocytes in human dental pulp repair. Reviewed

    Yoshiba N, Edanami N, Tohma A, Takeuchi R, Ohkura N, Hosoya A, Noiri Y, Nakamura H, Yoshiba K

    International endodontic journal   51 ( 11 )   1187 - 1195   2018.11

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    Aim: To explore the expression profile of CD45+/pro-collagen I+ fibrocytes in intact dental pulps as well as during wound healing in adult dental pulp tissue. Methodology: A total of 16 healthy permanent teeth were obtained from young patients (18 to 25 years) undergoing orthodontic treatment. Routine pulp capping with mineral trioxide aggregate (MTA) was performed under local anaesthesia to induce a mineralized barrier at the exposed surface. Teeth were extracted from patients after 7, 14 and 35 days. Sections of the extracted teeth were prepared and stained for various markers using indirect immunofluorescence. Fibrocytes were counted, and the data were statistically evaluated using the Dunnett test. Results: In uninflammed pulp tissue, a pro-collagen I-positive reaction was detected in odontoblasts, as well as in perivascular cells. Most of the CD45-positive cells were negative for pro-collagen I in normal pulp tissue, whereas CD45+/pro-collagen I+ fibrocytes were detected 7 days after injury. At day 14, fibrocytes were recognized under the fibrous matrix in contact with MTA and had infiltrated into regions of new capillary formation, where the fibrocytes were positively stained for vascular endothelial growth factor. By 35 days, fibrocytes were few, coincident with the formation of dentine bridges. The number of fibrocytes peaked 7 days post-injury and decreased at 14 days. Conclusions: The presence of fibrocytes in human pulp wound healing was observed. The spatiotemporal distribution of fibrocytes suggests that fibrocytes are involved in the early stages of pulp wound healing, specifically by contributing to new blood vessel formation.

    DOI: 10.1111/iej.12940

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  • ラットにおけるパルプ・リバスクラリゼーション治療実験モデルの開発 Reviewed

    白柏 麻里, 枝並 直樹, 吉羽 邦彦, 大倉 直人, 吉羽 永子, 遠間 愛子, 竹内 亮祐, 野杁 由一郎

    特定非営利活動法人日本歯科保存学会学術大会プログラムおよび講演抄録集   149回   120 - 120   2018.10

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  • ラット臼歯歯髄における創傷治癒時のアスコルビン酸輸送経路とその機能解析 Reviewed

    大倉 直人, 遠間 愛子, 竹内 亮祐, 枝並 直樹, 吉羽 永子, 吉羽 邦彦, 野杁 由一郎

    特定非営利活動法人日本歯科保存学会学術大会プログラムおよび講演抄録集   149回   33 - 33   2018.10

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  • 糖尿病モデルラットを用いた歯髄創傷治癒過程におけるCD68およびKi-67の発現解析 Reviewed

    遠間 愛子, 大倉 直人, 白柏 麻里, 竹内 亮祐, 枝並 直樹, 吉羽 永子, 吉羽 邦彦, 野杁 由一郎

    特定非営利活動法人日本歯科保存学会学術大会プログラムおよび講演抄録集   149回   103 - 103   2018.10

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  • 創傷治癒モデルラットを用いたSmad ubiquitination regulatory factor 1(SMURF1)およびmammalian target of rapamycin(mTOR)の免疫局在と遺伝子発現解析 Reviewed

    竹内 亮祐, 大倉 直人, 遠間 愛子, 白柏 麻里, 枝並 直樹, 吉羽 永子, 吉羽 邦彦, 野杁 由一郎

    特定非営利活動法人日本歯科保存学会学術大会プログラムおよび講演抄録集   149回   105 - 105   2018.10

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  • マイクロスコープを用いた再歯根尖切除術の1例

    大倉 直人, 山本 信一, 阿部 達也, 竹内 亮祐, 遠間 愛子, 枝並 直樹, 吉羽 永子, 吉羽 邦彦, 野杁 由一郎

    新潟歯学会雑誌   48 ( 1 )   29 - 35   2018.6

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    根尖性歯周組織病変の難治化は、根尖部での複雑な解剖学的問題に加え、残存バイオフィルムによる細菌学的な影響もあるとされている。こうした様々な要因の改善策として外科的歯内療法が選択される。しかし、補綴処置を行う上で考慮する歯冠・歯根長比の問題で複数回に渡り歯根切断が行われることは稀である。今回は、歯根尖切除術を過去3度に渡って施行しても治癒しなかった症例に対して、マイクロスコープの使用下で再歯根尖切除術と根尖部の緊密な封鎖を目的としたmineral trioxide aggregate(以下MTA)による逆根管充填を含めた再外科的歯内療法が奏効した症例を報告する。患者は59歳の女性で、上顎右側中切歯および側切歯に対して打診痛、根尖部圧痛および瘻孔を認める慢性根尖膿瘍と診断し、最初に通常の感染根管治療を行った。その後、病変部の治癒が期待できなかったため歯根尖切除を行い、逆根管形成後にMTAで逆根管充填を行った。手術から1ヵ月後で瘻孔は消失し、さらに6ヵ月後のレントゲン写真では根尖部の透過像が消失しており、治癒傾向を認めた。(著者抄録)

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  • Characterization of Dental Pulp Myofibroblasts in Rat Molars after Pulpotomy. Reviewed International journal

    Naoki Edanami, Nagako Yoshiba, Naoto Ohkura, Ryosuke Takeuchi, Aiko Tohma, Yuichiro Noiri, Kunihiko Yoshiba

    Journal of endodontics   43 ( 7 )   1116 - 1121   2017.7

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    INTRODUCTION: Myofibroblasts express alpha smooth muscle actin (α-SMA) and play a critical role in wound healing. Myofibroblast differentiation is controlled by the joint actions of transforming growth factor beta 1 (TGF-β1) and the extradomain A fibronectin splice variant (EDA-FN). Currently, the contribution of myofibroblasts to dental pulp healing is unknown. Therefore, we analyzed expressional characteristics of α-SMA-positive cells and investigated TGF-β1, EDA-FN, and α-SMA expression levels after pulpotomy to better understand dental pulp healing. METHODS: The maxillary first molars of 8-week-old Wistar rats were pulpotomized with mineral trioxide aggregate. After 1 to 14 days, localization and colocalization of α-SMA, rat endothelial cell antigen-1 (as a marker of endothelial cells), neuron-glial antigen 2 (as a marker of perivascular cells), prolyl-4-hydroxylase (P4H, as an additional marker of myofibroblasts), and EDA-FN were analyzed using immunohistochemistry and double immunofluorescence. Time-course changes in the messenger RNA expression levels of TGF-β1, EDA-FN, and α-SMA were evaluated using quantitative real-time polymerase chain reaction analysis. RESULTS: Spindle-shaped α-SMA-positive cells transiently appeared after pulpotomy. These cells initially emerged in the pulp core on day 3 and then accumulated at the wound site by day 5. These cells were isolated from rat endothelial cell antigen-1 positive cells and did not express neuron-glial antigen 2 but did express P4H. The messenger RNA levels of TGF-β1, EDA-FN, and α-SMA were significantly up-regulated after pulpotomy. EDA-FN and α-SMA were colocalized at the wound sites on day 5. CONCLUSIONS: In association with up-regulation of TGF-β1 and EDA-FN expression, α-SMA and P4H double-positive cells accumulated at the wound sites after pulpotomy. This suggests that myofibroblasts participate in dental pulp healing.

    DOI: 10.1016/j.joen.2017.02.018

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  • GaAlAs laser-induced pulp mineralization involves dentin matrix protein 1 and osteopontin expression Reviewed

    Y. Shigetani, N. Ohkura, K. Yoshiba, H. Ohshima, A. Hosoya, N. Yoshiba, T. Okiji

    Oral Diseases   22 ( 5 )   399 - 405   2016.7

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    Objectives: GaAlAs lasers induce pulp mineralization by promoting reparative dentinogenesis. This study analyzed the expression of dentin matrix protein 1 (DMP1) and osteopontin in GaAlAs laser-irradiated rat molars, to examine the hypothesis that these proteins play a role in the laser-induced reparative dentinogenic process. Materials and methods: The mesial surfaces of the upper first molars of 8-week-old Wistar rats were irradiated with a pulsed GaAlAs laser. After 1–14 days, mRNA expression of DMP1 and osteopontin in the coronal pulp was analyzed using real-time PCR. DMP1, osteopontin, and heat shock protein 25 (HSP25) were immunolocalized at 1–21 days. Results: The pulp exhibited a degenerative zone in its mesial portion on days 1–3, and progressive formation of reparative dentin lined with HSP25-immunoreactive odontoblast-like cells, from day 7 onwards. DMP1 and osteopontin mRNA expression were significantly upregulated on days 1–7 and 3–7, respectively. From day 7 onwards, DMP1 and osteopontin immunoreactivity colocalized along the boundary between the primary and reparative dentin. Conclusion: GaAlAs laser irradiation of rat molars induced upregulated DMP1 and osteopontin mRNA expression in the coronal pulp, followed by the formation of reparative dentin and the colocalization of DMP1 and osteopontin immunoreactivity at the site at which this tissue first appeared.

    DOI: 10.1111/odi.12461

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  • Correlation between Fibrillin-1 Degradation and mRNA Downregulation and Myofibroblast Differentiation in Cultured Human Dental Pulp Tissue. Reviewed International journal

    Nagako Yoshiba, Kunihiko Yoshiba, Naoto Ohkura, Erika Takei, Naoki Edanami, Youhei Oda, Akihiro Hosoya, Hiroaki Nakamura, Takashi Okiji

    The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society   63 ( 6 )   438 - 48   2015.6

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    Myofibroblasts and extracellular matrix are important components in wound healing. Alpha-smooth muscle actin (α-SMA) is a marker of myofibroblasts. Fibrillin-1 is a major constituent of microfibrils and an extracellular-regulator of TGF-β1, an important cytokine in the transdifferentiation of resident fibroblasts into myofibroblasts. To study the correlation between changes in fibrillin-1 expression and myofibroblast differentiation, we examined alterations in fibrillin-1 and α-SMA expression in organotypic cultures of dental pulp in vitro. Extracted healthy human teeth were cut to 1-mm-thick slices and cultured for 7 days. In intact dental pulp, fibrillin-1 was broadly distributed, and α-SMA was observed in pericytes and vascular smooth muscle cells. After 7 days of culture, immunostaining for fibrillin-1 became faint concomitant with a downregulation in its mRNA levels. Furthermore, fibroblasts, odontoblasts and Schwann cells were immunoreactive for α-SMA with a significant increase in α-SMA mRNA expression. Double immunofluorescence staining was positive for pSmad2/3, central mediators of TGF-β signaling, and α-SMA. The administration of inhibitors for extracellular matrix proteases recovered fibrillin-1 immunostaining; moreover, fibroblasts lost their immunoreactivity for α-SMA along with a downregulation in α-SMA mRNA. These findings suggest that the expression of α-SMA is TGF-β1 dependent, and fibrillin-1 degradation and downregulation might be implicated in the differentiation of myofibroblasts in dental pulp wound healing.

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  • ヒト歯髄創傷治癒過程における細胞外基質の局在変化 Fibrillin-1基質の動的リモデリングに関する検索

    吉羽 永子, 吉羽 邦彦, 大倉 直人, 重谷 佳見, 武井 絵梨花, 細矢 明宏, 中村 浩彰, 興地 隆史

    日本歯科保存学雑誌   56 ( 3 )   161 - 168   2013.6

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    目的:細胞外基質fibrillin-1は,弾性線維の構成タンパクであると同時に,その分解によりtransforming growth factor-β(TGF-β)を遊離させることが知られている.また,著者らはfibrillin-1がヒト歯髄直接覆髄後の創傷治癒過程で発現を消失させることを見いだしている.そこで本研究では,TGF-β制御活性を有する細胞外基質decorinとfibronectin,および創傷治癒と組織線維化に関与するtenascin-Cが,ヒト歯髄直接覆髄後の創傷治癒過程でfibrillin-1様の局在変化を示すか否かを検索した.さらに,スライス培養したヒト歯髄を用いて,fibrillin-1の発現消失へのmatrix metalloproteinase(MMP)の関与の実態を検索した.材料と方法:矯正治療により抜歯予定のヒト健全歯をmineral trioxide aggregate(MTA)で直接覆髄した後,2週および6週後に抜歯し,fibrillin-1,fibronectin,decorinおよびtenascin-Cに対する免疫組織化学的染色を行った.また,健全歯髄を組織培養し,fibrillin-1 mRNA発現を定量RT-PCRで測定するとともに,MMP阻害剤であるNNGHを添加培養し,fibrillin-1タンパクの局在変化を免疫染色により解析した.結果:直接覆髄後2週および6週において,fibrillin-1に対する免疫陽性反応は,覆髄部直下の歯髄組織で局所的に消失していた.一方,fibronectin,decorinおよびtenascin-Cは同部で一様に陽性反応を示しており,明らかな局在変化は認められなかった.一方,培養歯髄組織ではfibrillin-1の染色性が減弱し,mRNAの発現も有意に低下したが,MMP-3 mRNA発現は有意に亢進した.NNGHを添加培養するとfibrillin-1の染色性が向上した.結論:検索対象とした4種のタンパクのなかでfibrillin-1のみ直接覆髄後の創傷治癒過程で局在変化を示した.Fibrillin-1免疫反応性の消失には,MMPsによるタンパクの分解に加えて遺伝子発現の下方制御も関与するものと考えられた.(著者抄録)

    DOI: 10.11471/shikahozon.56.161

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  • Immunohistochemical analysis of two stem cell markers of α-smooth muscle actin and STRO-1 during wound healing of human dental pulp. Reviewed International journal

    Nagako Yoshiba, Kunihiko Yoshiba, Naoto Ohkura, Yoshimi Shigetani, Erika Takei, Akihiro Hosoya, Hiroaki Nakamura, Takashi Okiji

    Histochemistry and cell biology   138 ( 4 )   583 - 92   2012.10

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    Recent studies have employed two markers, alpha-smooth muscle actin (α-SMA) and STRO-1, to detect cells with mesenchymal stem cell properties in dental pulp. The present study aimed to explore the expression profile of α-SMA and STRO-1 in intact dental pulp as well as during wound healing in adult dental pulp tissue. Healthy pulps were mechanically exposed and capped with the clinically used materials MTA (ProRoot White MTA) or Ca(OH)₂ to induce a mineralized barrier at the exposed surface. After 7-42 days, the teeth were extracted and processed for immunohistochemical analysis using antibodies against α-SMA, STRO-1 and nestin (a neurogenic cytoskeletal protein expressed in odontoblasts). In normal pulp, α-SMA was detected in vascular smooth muscle cells and pericytes. Double immunofluorescent staining with STRO-1 and α-SMA showed that STRO-1 was localized in vascular smooth muscle cells, pericytes and endothelial cells, in addition to nerve fibers. During the process of dental pulp healing, numerous α-SMA-positive cells emerged at the wound margin at 14 days, and the initially formed mineralized barrier was lined with α-SMA-positive cells similar in appearance to reparative odontoblasts, some of which co-expressed nestin. STRO-1 was abundant in nerve fibers. In the advanced stage of mineralized barrier formation at 42 days, cells lining the barrier were stained with nestin, and no staining of α-SMA was detected in those cells. These observations indicate that α-SMA-positive cells temporarily appear along the wound margin during the earlier phase of mineralized barrier formation and STRO-1 is confined in vascular and neuronal elements.

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  • Gene expression analysis of membrane transport proteins in normal and lipopolysaccharide-inflamed rat dental pulp. Reviewed International journal

    Naoto Ohkura, Yoshimi Shigetani, Nagako Yoshiba, Kunihiko Yoshiba, Takashi Okiji

    Journal of endodontics   38 ( 5 )   648 - 52   2012.5

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    INTRODUCTION: Membrane transport proteins (transporters) play a crucial role in the transmembrane uptake and/or efflux of various compounds such as inorganic ions, endogenous bioactive substances such as prostaglandins (PGs), and drugs such as nonsteroidal anti-inflammatory drugs. This study aimed to analyze mRNA expression of selected transporters related to drug disposition and PG transport in normal and lipopolysaccharide (LPS)-inflamed rat incisor pulp. METHODS: Pulp tissues were subjected to reverse transcription-polymerase chain reaction (PCR) detection for transporter isoforms belonging to organic anion transporting polypeptide (Oatp), organic anion transporter (Oat), organic cation transporter (Oct), multidrug resistance-associated protein (Mrp), and multidrug resistance protein (Mdr) families. The levels of mRNA expression for PG transporters (Oatp1a5, Oatp1b2, Oatp2a1, Oatp2b1, and Oatp3a1) were compared in normal and LPS-inflamed pulps by using real-time PCR. RESULTS: The pulp tissue expressed mRNAs for various transporters belonging to the Oatp, Oat, Oct, Mrp, and Mdr families. LPS inflammation caused significant up-regulation of Oatp2a1 (P < .01) and significant down-regulation of Oatp1a5, Oatp2b1 (P < .01), and Oatp3a1 (P < .05). CONCLUSIONS: Rat incisor dental pulp expressed mRNAs for various transporter isoforms. The levels of mRNA expression for PG transporters were significantly up-regulated or down-regulated in LPS-inflamed dental pulp.

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  • Expressional alterations of fibrillin-1 during wound healing of human dental pulp. Reviewed International journal

    Nagako Yoshiba, Kunihiko Yoshiba, Naoto Ohkura, Akihiro Hosoya, Yoshimi Shigetani, Yusuke Yamanaka, Naoya Izumi, Hiroaki Nakamura, Takashi Okiji

    Journal of endodontics   38 ( 2 )   177 - 84   2012.2

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    INTRODUCTION: The degradation of fibrillins, the major constituents of microfibrils, is known to facilitate the release of active transforming growth factor-β (TGF-β), a signaling molecule contributing to mineralized tissue barrier formation in exposed dental pulps. To examine the involvement of fibrillins in the barrier formation, we examined the temporospatial expression of (1) genes and proteins of fibrillins and (2) factors possibly associated with fibrillin degradation and cytodifferentiation in exposed human pulps. Human pulp slice cultures were also examined for the role of fibrillins in mineralization. METHODS: Clinically healthy pulps were mechanically exposed and capped with mineral trioxide aggregate. After 7 to 42 days, the teeth were processed for immunohistochemical and cytochemical staining of fibrillin-1, fibrillin-2, latent TGF-β-binding protein (LTBP)-1, matrix metalloproteinase-3 (MMP-3), alkaline phosphatase (ALP), and in situ hybridization of fibrillin-1. Pulp tissue slices cultured with β-glycerophosphate were analyzed for fibrillin-1, fibrillin-2, and ALP with the immunohistochemical/cytochemical staining and quantitative reverse-transcriptase polymerase chain reaction. RESULTS: Fibrillin-1-immunoreactivity was seen until 7 days but turned into undetectable since 14 days in the pulpal area just beneath the exposure site. MMP-3-immunoreaction was transiently detected at 14 days. At 42 days when the mineralized barrier was evident, fibrillin-1-immunoreactivity and fibrillin-1 expression remained down-regulated. Fibrillin-2, LTBP-1, and ALP were constantly detected in the fibrillin-1-undetectable area. Pulp slices cultured with β-glycerophosphate showed mineralization with up-regulation of ALP and down-regulation of fibrillin-1. CONCLUSIONS: Degradation and down-regulation of fibrillin-1 expression took place during the mineralized tissue barrier formation in exposed pulps in vivo and β-glycerophosphate-induced pulpal mineralization in vitro.

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  • 半導体レーザー照射後のラット臼歯における非コラーゲンタンパクの遺伝子発現

    重谷 佳見, 大倉 直人, 吉羽 邦彦, 吉羽 永子, 大島 勇人, 興地 隆史

    日本歯科保存学雑誌   53 ( 5 )   495 - 501   2010.10

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    ラット臼歯へGaAlAs半導体レーザーを照射した後に,歯髄腔内に硬組織形成が生じることが知られている.そこで本研究では,RT-PCR法を用いて同レーザー照射後のラット臼歯における各種非コラーゲンタンパクのmRNA発現を検索した.生後8週齢Wistar系雄性ラットの上顎第一臼歯近心に,半導体レーザー装置(オサダライトサージ3000)を用いて,出力1.5W,60秒3回の条件でレーザー照射を行った.なお,非照射の上顎臼歯を対照とした.照射1,3,7日後に抜歯を行い,歯冠部よりmRNAを抽出した後,RT-PCR法にてosteo-pontin,osteonectin,osteocalcin,dentin sialophosphoprotein,dentin matrix protein 1,およびbone sialoproteinのmRNA発現解析を行った.また,歯髄の反応を組織学的に観察した.その結果,すべてのタンパクのmRNA発現が3日後までに増加を示すとともに,この発現亢進は7日後も持続していることが明らかになった.組織学的には,1〜3日後までは歯髄の局所的壊死が照射部近傍に観察されたが,7日後には象牙芽細胞様細胞の再配列と少量の新生硬組織が認められた.以上より,半導体レーザー照射されたラット臼歯では,新生硬組織形成に先立ち各種非コラーゲンタンパクのmRNA発現レベルの上昇が生じることが確認された.(著者抄録)

    DOI: 10.11471/shikahozon.53.495

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  • Tie2-EGFPトランスジェニックマウスを用いた高純度脳毛細血管内皮細胞単離(Fluorescence activated purification of brain capillary endothelial cells from Tie2-EGFP transgenic mouse)

    寺崎 哲也, 大倉 直人, 堀 里子, 大槻 純男

    神経化学   43 ( 2-3 )   431 - 431   2004.8

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  • Impact of the tissue nonspecific alkaline phosphatase during the root formation

    大倉直人, 大倉直人, 吉羽永子, 高原信太郎, GUTIERREZ Rosa Edith Baldeon, GOMEZ-KASIMOTO Susan, 井田貴子, 枝並直樹, 竹中彰治, 吉羽邦彦, 野杁由一郎

    日本歯科保存学会学術大会プログラムおよび講演抄録集(Web)   159th   92 - 92   2023

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  • 専門医への道 症例報告

    大倉 直人

    日本歯内療法学会雑誌   40 ( 2 )   117 - 120   2019.5

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    症例は72歳男性で、数日前から上顎左側犬歯の根尖部に違和感をきたしていた。デンタルX線写真では根尖周囲において広範囲のX線透過像を認め、歯根近心部では歯根膜腔の拡大と思われる所見を認めた。初回治療として、Kファイルで慎重に根尖孔の穿通を行い、2%次亜塩素酸ナトリウムと3%EDTA溶液を用いて根管に対して交互洗浄を念入りに行った。2回目治療として、側方加圧充填法、根管充填用シーラーによって根管充填した。3回目治療として、ラバーダム防湿後、フロアブルレジンを用いてコンポジットレジンによる歯冠部修復を行った。根管充填4ヵ月後、自覚・他覚症状はともに認められず、デンタルX線所見においては骨吸収像と推定される初診時に認められた根尖周囲の透過像が改善していた。根管充填6ヵ月後、自覚・他覚症状はともに認められず、デンタルX線所見で病変の再発を示す所見はみられず良好に経過していた。

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  • マイクロスコープを用いた再歯根尖切除術の1例

    大倉 直人, 山本 信一, 阿部 達也, 竹内 亮祐, 遠間 愛子, 枝並 直樹, 吉羽 永子, 吉羽 邦彦, 野杁 由一郎

    新潟歯学会雑誌   48 ( 1 )   29 - 35   2018.6

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    根尖性歯周組織病変の難治化は、根尖部での複雑な解剖学的問題に加え、残存バイオフィルムによる細菌学的な影響もあるとされている。こうした様々な要因の改善策として外科的歯内療法が選択される。しかし、補綴処置を行う上で考慮する歯冠・歯根長比の問題で複数回に渡り歯根切断が行われることは稀である。今回は、歯根尖切除術を過去3度に渡って施行しても治癒しなかった症例に対して、マイクロスコープの使用下で再歯根尖切除術と根尖部の緊密な封鎖を目的としたmineral trioxide aggregate(以下MTA)による逆根管充填を含めた再外科的歯内療法が奏効した症例を報告する。患者は59歳の女性で、上顎右側中切歯および側切歯に対して打診痛、根尖部圧痛および瘻孔を認める慢性根尖膿瘍と診断し、最初に通常の感染根管治療を行った。その後、病変部の治癒が期待できなかったため歯根尖切除を行い、逆根管形成後にMTAで逆根管充填を行った。手術から1ヵ月後で瘻孔は消失し、さらに6ヵ月後のレントゲン写真では根尖部の透過像が消失しており、治癒傾向を認めた。(著者抄録)

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  • 神経を抜かない新しいむし歯(う蝕)治療の開発

    大倉直人, 大倉直人, 野杁由一郎, 野杁由一郎

    Bio Industry   35 ( 10 )   2018

  • 歯髄創傷治癒のメカニズム-プロスタグランジンE<sub>2</sub>を中心に-

    大倉直人, 野杁由一郎

    日本歯内療法学会雑誌   39 ( 3 )   2018

  • 培養ヒト歯髄に対するprostaglandin EP4レセプターアゴニストの影響

    大倉 直人, 吉羽 永子, 吉羽 邦彦, 小田 陽平, 興地 隆史

    日本歯科医師会雑誌   69 ( 5 )   454 - 454   2016.8

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  • マイクロスコープを用いた再歯根尖切除術の1症例

    大倉 直人, 山本 信一, 枝並 直樹, 吉羽 永子, 吉羽 邦彦, 野杁 由一郎

    日本歯内療法学会学術大会プログラム・抄録集   37回   60 - 60   2016.6

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  • 培養ヒト歯髄に対するprostaglandin EP2レセプターアゴニストの影響

    大倉 直人, 枝並 直樹, 竹内 亮祐, 遠間 愛子, 吉羽 永子, 吉羽 邦彦, 小田 陽平, 興地 隆史

    特定非営利活動法人日本歯科保存学会学術大会プログラムおよび講演抄録集   144回   124 - 124   2016.6

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  • 培養ヒト歯髄に対するprostaglandin EP4レセプターアゴニストの影響

    大倉 直人, 吉羽 永子, 吉羽 邦彦, 小田 陽平, 興地 隆史

    特定非営利活動法人日本歯科保存学会学術大会プログラムおよび講演抄録集   143回   75 - 75   2015.11

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  • ラット臼歯歯髄断髄後のProstaglandin Transporterに対する免疫組織学的局在解析

    大倉 直人, 枝並 直樹, 吉羽 永子, 吉羽 邦彦, 依田 浩子, 大島 勇人, 興地 隆史

    Journal of Oral Biosciences Supplement   2015   367 - 367   2015.9

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  • 培養ヒト歯髄の各種遺伝子発現に対するprostaglandin EP4レセプターアゴニストの影響

    大倉 直人, 吉羽 永子, 吉羽 邦彦, 興地 隆史

    日本歯内療法学会学術大会プログラム・抄録集   36回   71 - 71   2015.7

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  • ラット臼歯歯髄断髄後のProstaglandin Transporterに対する免疫組織学的局在解析

    大倉直人, 枝並直樹, 吉羽永子, 吉羽邦彦, 依田浩子, 大島勇人, 興地隆史

    Journal of Oral Biosciences Supplement (Web)   2015   2015

  • ヒト歯髄におけるプロスタグランジンE2輸送担体および特異的レセプターの免疫組織化学的局在解析

    大倉 直人, 大倉 麻里子, 吉羽 永子, 吉羽 邦彦, 小田 陽平, 依田 浩子, 大島 勇人, 興地 隆史

    Journal of Oral Biosciences Supplement   2014   183 - 183   2014.9

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  • ヒト歯髄におけるProstaglandinレセプターの免疫組織学的局在解析

    大倉 直人, 重谷 佳見, 吉羽 永子, 吉羽 邦彦, 興地 隆史

    日本歯内療法学会学術大会プログラム・抄録集   35回   101 - 101   2014.7

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  • ヒト歯髄におけるプロスタグランジンE<sub>2</sub>輸送担体および特異的レセプターの免疫組織化学的局在解析

    大倉直人, 大倉麻里子, 吉羽永子, 吉羽邦彦, 小田陽平, 依田浩子, 大島勇人, 興地隆史

    Journal of Oral Biosciences Supplement (Web)   2014   2014

  • ラット炎症歯髄に対する薬物輸送担体を介したProstaglandin E2輸送経路解析

    大倉 直人, 重谷 佳見, 吉羽 永子, 吉羽 邦彦, 興地 隆史

    新潟歯学会雑誌   43 ( 2 )   155 - 155   2013.12

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  • ラット臼歯矯正移動時における歯髄内prostanoid receptorの遺伝子発現と免疫組織学的局在解析

    大倉 直人, 大倉 麻里子, 重谷 佳見, 吉羽 永子, 吉羽 邦彦, 齋藤 功, 興地 隆史

    Journal of Oral Biosciences Supplement   2013   189 - 189   2013.9

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  • ラット臼歯矯正移動時における歯髄内prostanoid receptorの遺伝子発現と免疫組織学的局在解析

    大倉直人, 大倉麻里子, 重谷佳見, 吉羽永子, 吉羽邦彦, 齋藤功, 興地隆史

    Journal of Oral Biosciences Supplement (Web)   2013   ROMBUNNO.P2‐24 (WEB ONLY)   2013

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  • ラット炎症歯髄に対する薬物輸送担体の遺伝子発現解析

    大倉 直人, 重谷 佳見, 細矢 明宏, 吉羽 永子, 吉羽 邦彦, 興地 隆史

    Journal of Oral Biosciences Supplement   2012   160 - 160   2012.9

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  • ラット炎症歯髄に対する薬物輸送担体の遺伝子発現解析

    大倉 直人, 重谷 佳見, 細矢 明宏, 吉羽 永子, 吉羽 邦彦, 興地 隆史

    日本歯科医師会雑誌   65 ( 5 )   628 - 628   2012.8

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  • ラット実験的炎症歯髄におけるプロスタノイド受容体遺伝子発現の経時的解析

    大倉 直人, 重谷 佳見, 細矢 明宏, 吉羽 永子, 吉羽 邦彦, 興地 隆史

    特定非営利活動法人日本歯科保存学会学術大会プログラムおよび講演抄録集   134回   104 - 104   2011.5

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  • ラット炎症歯髄に対する薬物輸送担体の遺伝子発現解析

    大倉 直人, 重谷 佳見, 細矢 明宏, 吉羽 永子, 吉羽 邦彦, 興地 隆史

    特定非営利活動法人日本歯科保存学会学術大会プログラムおよび講演抄録集   133回   96 - 96   2010.10

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  • 頬部粘膜腫瘍摘出後に発症した急性閉塞隅角緑内障の1例

    大倉直人, 吉川博之, 塚田博子, 倉田行伸, 弦巻立, 田中裕, 瀬尾憲司, 染矢源治

    関東臨床歯科麻酔懇話会   26th   2009

  • Tie2-EGFPマウスを用いた高純度脳毛細血管内皮細胞単離法の開発

    大倉 直人, 堀 里子, 大槻 純男, 寺崎 哲也

    日本薬学会年会要旨集   124年会 ( 4 )   78 - 78   2004.3

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  • Sphingosine-1-phosphate (S1P) receptor 1-mediated odontoblastic differentiation and mineralization of mouse apical papilla-derived stem cells

    廣瀬陽菜, 藤政清志朗, 金丸慎吾, 松本典祥, 高原信太郎, 大倉直人, 枝並直樹, 野杁由一郎, 松崎英津子, 松崎英津子

    日本歯科保存学会学術大会プログラムおよび講演抄録集(Web)   159th   46 - 46   2023

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  • Development of Point of Care Testing to facilitate risk assessment for the elderly at home-Performance evaluation of a newly developed semi-quantitative rapid test for CRP level determination-

    竹中彰治, 枝並直樹, 齋藤瑠郁, 大倉直人, 野杁由一郎

    日本歯科保存学会学術大会プログラムおよび講演抄録集(Web)   158th   148 - 148   2023

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  • The localization of various mesenchymal stem cell marker-positive cells in early healing stage after regenerative endodontic procedure

    高原信太郎, 大倉直人, 吉羽永子, 竹中彰治, 枝並直樹, 吉羽邦彦, 野杁由一郎

    日本歯科保存学会学術大会プログラムおよび講演抄録集(Web)   159th   47 - 47   2023

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  • The comparison analyses of wound-healing mechanism according to the root development stage using the regeneration model rats

    高原信太郎, 大倉直人, 吉羽邦彦, 吉羽永子, 竹中彰治, 枝並直樹, 庭野和明, 野杁由一郎

    日本歯科保存学会学術大会プログラムおよび講演抄録集(Web)   158th   31 - 31   2023

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  • Interactions between Apically-extruded Bioceramic Sealers and periodontal tissues

    高原信太郎, 枝並直樹, 竹中彰治, 吉羽邦彦, 大倉直人, 吉羽永子, 野杁由一郎

    日本歯科保存学会学術大会プログラムおよび講演抄録集(Web)   157回   125 - 125   2022.10

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  • Comparison of Biomineralization Ability of Calcium Silicate-based and Calcium Hydroxide-based Intracanal Medicaments

    枝並直樹, 竹中彰治, 吉羽邦彦, 大倉直人, 吉羽永子, 高原信太郎, 野杁由一郎

    日本歯科保存学会学術大会プログラムおよび講演抄録集(Web)   157回   126 - 126   2022.10

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  • ケイ酸カルシウム系貼薬剤と水酸化カルシウム系貼薬剤のBiomineralization Abilityの比較

    枝並 直樹, 竹中 彰治, 吉羽 邦彦, 大倉 直人, 吉羽 永子, 高原 信太郎, 野杁 由一郎

    特定非営利活動法人日本歯科保存学会学術大会プログラムおよび講演抄録集   157回   126 - 126   2022.10

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  • 根尖孔外に漏出したバイオセラミック系シーラーと歯周組織の相互作用

    高原 信太郎, 枝並 直樹, 竹中 彰治, 吉羽 邦彦, 大倉 直人, 吉羽 永子, 野杁 由一郎

    特定非営利活動法人日本歯科保存学会学術大会プログラムおよび講演抄録集   157回   125 - 125   2022.10

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  • Application of Autologous Concentrated Growth Factors (CGF) in Endodontic Microsurgery: A Multi-Center Clinical Trial

    八幡祥生, 半田慶介, 半田慶介, 大倉直人, 伊藤祥作, 川島伸之, 野杁由一郎, 林美加子, 興地隆史, 齋藤正寛

    日本歯科医学会誌   40   43 - 48   2021.3

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  • ヒト歯髄においてシュワン細胞はマクロファージをM2型へ転換する

    吉羽永子, 大倉直人, 前川知樹, 泉健次, 細矢明宏, 中村浩彰, 前田健康, 野杁由一郎, 吉羽邦彦

    Journal of Oral Biosciences Supplement (Web)   2019   2019

  • 歯髄創傷モデルラットを用いた修復象牙質形成時におけるGlut1-Runx2関連の解析

    竹内 亮祐, 大倉 直人, 枝並 直樹, 遠間 愛子, 吉羽 永子, 吉羽 邦彦, 野杁 由一郎

    新潟歯学会雑誌   48 ( 2 )   113 - 114   2018.12

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  • 歯髄創傷治癒モデルラットを用いたグルコース輸送担体Glut2とGlut4の局在および遺伝子発現の解析

    遠間 愛子, 大倉 直人, 枝並 直樹, 竹内 亮祐, 吉羽 永子, 吉羽 邦彦, 野杁 由一郎

    新潟歯学会雑誌   48 ( 2 )   114 - 114   2018.12

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  • ODSラットを用いた矯正的歯の移動におけるアスコルビン酸の影響

    大倉 麻里子, 大倉 直人, 丹原 惇, 中田 樹里, 藤田 瑛, 野杁 由一郎, 齋藤 功

    日本矯正歯科学会大会プログラム・抄録集   77回   232 - 232   2018.10

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  • 歯髄創傷治癒過程におけるマクロファージの集積とmyofibroblast様細胞の分化

    枝並 直樹, 吉羽 邦彦, 吉羽 永子, 大倉 直人, 竹内 亮祐, 遠間 愛子, 野杁 由一郎

    特定非営利活動法人日本歯科保存学会学術大会プログラムおよび講演抄録集   147回   182 - 182   2017.10

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  • 矯正的歯の移動におけるprostaglandin I2合成酵素の発現解析と3次元的応力分布解析

    大倉 麻里子, 大倉 直人, 丹原 惇, 藤田 瑛, 野杁 由一郎, 齋藤 功

    日本矯正歯科学会大会プログラム・抄録集   76回   183 - 183   2017.10

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  • 歯髄創傷治癒モデルラットを用いたGlucose Transporter-4の局在および遺伝子発現の解析

    遠間 愛子, 大倉 直人, 枝並 直樹, 竹内 亮祐, 吉羽 永子, 吉羽 邦彦, 野杁 由一郎

    特定非営利活動法人日本歯科保存学会学術大会プログラムおよび講演抄録集   147回   46 - 46   2017.10

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  • 歯髄創傷治癒モデルラットを用いたGlucose Transporter-1およびrunt-related transcription factor 2の局在と遺伝子発現解析

    竹内 亮祐, 大倉 直人, 枝並 直樹, 遠間 愛子, 吉羽 永子, 吉羽 邦彦, 野杁 由一郎

    特定非営利活動法人日本歯科保存学会学術大会プログラムおよび講演抄録集   147回   86 - 86   2017.10

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  • ヒト歯髄組織創傷治癒過程における骨髄由来間葉系前駆細胞fibrocyteの動態検索

    吉羽 永子, 大倉 直人, 細矢 明宏, 中村 浩彰, 野杁 由一郎, 吉羽 邦彦, 枝並 直樹, 遠間 愛子, 竹内 亮介

    Journal of Oral Biosciences Supplement   2017   427 - 427   2017.9

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  • ラット臼歯断髄後におけるmyofibroblastの動態解析

    枝並 直樹, 吉羽 永子, 大倉 直人, 野杁 由一郎, 吉羽 邦彦

    新潟歯学会雑誌   47 ( 1 )   52 - 52   2017.6

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  • ヒト歯髄組織創傷治癒過程における骨髄由来間葉系前駆細胞fibrocyteの動態検索

    吉羽永子, 大倉直人, 細矢明宏, 中村浩彰, 野杁由一郎, 吉羽邦彦

    Journal of Oral Biosciences Supplement (Web)   2017   2017

  • 実験的歯の移動時におけるラット臼歯歯髄内prostaglandin I2合成酵素とIP受容体の発現解析

    大倉 麻里子, 大倉 直人, 野杁 由一郎, 齋藤 功

    日本矯正歯科学会大会プログラム・抄録集   75回   203 - 203   2016.11

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  • 各種ケイ酸カルシウム系セメントの生体機能性と直接覆髄後の歯髄反応

    吉羽 邦彦, 枝並 直樹, 日向 剛, 韓 臨麟, 竹内 亮祐, 遠間 愛子, 大倉 直人, 武井 絵梨花, 吉羽 永子, 興地 隆史

    日本歯科医師会雑誌   69 ( 5 )   454 - 454   2016.8

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  • ラット臼歯におけるMineral trioxide aggregateによる直接覆髄後のGlucose Transporter-1の免疫局在および遺伝子発現解析

    竹内 亮祐, 大倉 直人, 枝並 直樹, 遠間 愛子, 吉羽 永子, 吉羽 邦彦

    特定非営利活動法人日本歯科保存学会学術大会プログラムおよび講演抄録集   144回   112 - 112   2016.6

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  • ラット臼歯におけるMTAによる直接覆髄後のGlucose Transporter-2の免疫組織化学および遺伝子発現の解析

    遠間 愛子, 大倉 直人, 枝並 直樹, 竹内 亮祐, 吉羽 永子, 吉羽 邦彦

    特定非営利活動法人日本歯科保存学会学術大会プログラムおよび講演抄録集   144回   111 - 111   2016.6

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  • 実験的歯の移動におけるラット臼歯歯髄内prostaglandin I2合成酵素と受容体の発現解析

    大倉 麻里子, 大倉 直人, 吉羽 永子, 吉羽 邦彦, 依田 浩子, 大島 勇人, 齋藤 功, 興地 隆史

    新潟歯学会雑誌   45 ( 2 )   97 - 98   2015.12

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  • 矯正移動時におけるラット臼歯歯髄内prostaglandin I2受容体の発現解析

    大倉 麻里子, 大倉 直人, 興地 隆史, 齋藤 功

    日本矯正歯科学会大会プログラム・抄録集   73回   208 - 208   2014.10

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  • ヒト歯髄組織からoutgrowthする細胞による組織構築に関する研究

    吉羽 永子, 吉羽 邦彦, 大倉 直人, 細矢 明宏, 中村 浩彰, 興地 隆史

    Journal of Oral Biosciences Supplement   2013   193 - 193   2013.9

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  • ヒト歯髄組織からoutgrowthする細胞による組織構築に関する研究

    吉羽永子, 吉羽邦彦, 大倉直人, 細矢明宏, 中村浩彰, 興地隆史

    Journal of Oral Biosciences Supplement (Web)   2013   2013

  • 半導体レーザー照射後のラット臼歯における硬組織形成誘導機構の解明

    重谷 佳見, 大倉 直人, 細矢 明宏, 鈴木 啓展, 吉羽 邦彦, 吉羽 永子, 大島 勇人, 興地 隆史

    日本歯科医師会雑誌   65 ( 5 )   629 - 629   2012.8

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  • 半導体レーザー照射後のラット臼歯における硬組織関連タンパクの遺伝子発現

    重谷 佳見, 大倉 直人, 吉羽 邦彦, 吉羽 永子, 興地 隆史

    日本歯内療法学会学術大会プログラム・抄録集   31回   110 - 110   2010.5

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  • ヒト歯髄創傷治癒過程で生じるFibrillin-1の分解は細胞分化と石灰化を誘導する

    吉羽 永子, 吉羽 邦彦, 大倉 直人, 細矢 明宏, 重谷 佳見, 興地 隆史

    特定非営利活動法人日本歯科保存学会学術大会プログラムおよび講演抄録集   132回   117 - 117   2010.5

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  • LC-MSMS-Cocktail法を用いたhuman MRP4基質のhigh throughput screening法の開発

    内田 康雄, 上家 潤一, 大槻 純男, 矢上 礼宣, 藪内 光, 大倉 直人, 登美 斉俊, 細谷 健一, 寺崎 哲也

    日本薬学会年会要旨集   126年会 ( 2 )   115 - 115   2006.3

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  • Tie2/GFP導入トランスジェニックラットにおける血液脳及び網膜関門選択的な遺伝子発現

    大槻 純男, 大倉 直人, 紙谷 尚子, 堀 里子, 寺崎 哲也

    日本薬学会年会要旨集   125年会 ( 2 )   113 - 113   2005.3

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  • 蛍光プローブを用いたヒトABCC4の基質解析系の確立

    大槻 純男, Madarina Miron, 堀 里子, 大倉 直人, 登美 斉俊, 細谷 健一, 寺崎 哲也

    薬剤学: 生命とくすり   65 ( Suppl. )   142 - 142   2005.3

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  • 疎水環境変化を利用したヒトABCC4の基質スクリーニング法

    Madarina Miron, 大槻 純男, 堀 里子, 大倉 直人, 登美 斉俊, 細谷 健一, 寺崎 哲也

    日本薬学会年会要旨集   125年会 ( 2 )   115 - 115   2005.3

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Presentations

  • 夢の虫歯治療〜削らないむし歯治療の現状〜 Invited

    大倉直人

    日本歯科保存学会 市民公開フォーラム  2019.9 

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    Event date: 2019.9

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Awards

  • 優秀ポスター賞

    2024.5   日本歯科保存学会  

    大倉 直人

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  • 日本歯科保存学会奨励賞

    2018   日本歯科保存学会  

    大倉 直人

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  • デンツプライ賞

    2011   日本歯科保存学会  

    大倉 直人

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Research Projects

  • 歯髄の創傷治癒・再生過程におけるGli1陽性幹細胞の動態と分化誘導機構の解明

    Grant number:21K09914

    2021.4 - 2024.3

    System name:科学研究費助成事業

    Research category:基盤研究(C)

    Awarding organization:日本学術振興会

    吉羽 邦彦, 吉羽 永子, 枝並 直樹, 細矢 明宏, 入江 一元

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    Grant amount:\4160000 ( Direct Cost: \3200000 、 Indirect Cost:\960000 )

    歯の保存には歯髄の保護・保存が重要であり、直接覆髄や一部生活歯髄切断など歯髄保存療法が、また失われた歯髄に対しては再生歯内療法が試みられている。しかし根管内に形成される硬組織はセメント質あるいは骨様組織であり、理想的な「象牙質・歯髄複合体の再生」には至っていない。また、より迅速かつ確実な処置法の開発が望まれている。本研究では、歯髄の創傷治癒・再生過程において中心的役割を果たすと考えられている間葉系幹細胞の動態について、転写因子Gli1発現をマーカーとして細胞系譜解析により検索するとともに、覆髄材・根管充填材として注目されているバイオセラミックス配合材料の歯髄創傷治癒に与える影響について検討した。
    遺伝子改変(Gli1-CreERT2/Tomato)マウスにおいて、Gli1陽性細胞は歯根膜および歯髄の血管周囲に散在性に認められること、また、臼歯歯根膜から分取されたGli1陽性細胞はコロニー形成能、多分化能を示し幹細胞特性を持つことが明らかにされた。これらの結果から、再生歯内療法応用後の治癒過程、特に硬組織形成におけるGli1陽性細胞の関与が示唆された。一方、バイオセラミックス配合覆髄材・根管充填材のin vitro・in vivoにおけるアパタイト析出能を検討した結果、すべての被験材料は疑似体液中でその表面にCa-Pを含むアパタイト様構造物を析出させたが、ラット皮下移植実験では、材料間にその析出能の差異が認められた。また、レジン含有ケイ酸カルシウム(MTA)系セメントは断髄後、CD68陽性マクロファージの持続的な浸潤と修復象牙質形成の遅延が観察され、材料選択の重要性が示唆された。

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  • Comprehensive study on establishment of pulp regenerative therapy based on the scientific evidences

    Grant number:21H03117

    2021.4 - 2024.3

    System name:Grants-in-Aid for Scientific Research

    Research category:Grant-in-Aid for Scientific Research (B)

    Awarding organization:Japan Society for the Promotion of Science

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    Grant amount:\17550000 ( Direct Cost: \13500000 、 Indirect Cost:\4050000 )

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  • ヒト歯髄の創傷治癒過程におけるM2マクロファージとシュワン細胞の相互作用の解明

    Grant number:19K10146

    2019.4 - 2022.3

    System name:科学研究費助成事業 基盤研究(C)

    Research category:基盤研究(C)

    Awarding organization:日本学術振興会

    吉羽 永子, 吉羽 邦彦, 大倉 直人, 枝並 直樹

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    Grant amount:\4290000 ( Direct Cost: \3300000 、 Indirect Cost:\990000 )

    免疫機能の中心的役割を担っているマクロファージは、その誘導因子により、異なる性質を持つ2つの分化型に分化することが知られている。すなわちM1マクロファージと称される炎症を促進するものと、その反対にM2マクロファージという炎症を抑制し組織の修復に働くタイプのものである。本研究では、ヒト歯髄組織におけるマクロファージの特にM2型の動態について、in vivoおよびin vitroにおいて解析している。
    令和元年度では先ずin vivoすなわち、(1)ヒトの健全歯髄組織、(2)覆髄剤であるMTAを用いた直接覆髄後の創傷治癒過程、さらに(3)様々な病態のう蝕歯髄におけるM2マクロファージの動態を検索した。その結果、これら全ての状態において、CD163をマーカーとするM2マクロファーは神経線維保護機能を有するシュワン細胞と共在していることが明らかとなった。シュワン細胞はp75 nerve growth factor receptor (NGFR)を発現していることから、そのリガンドの1つであるbrain-derived neurotrophic factor (BDNF)の局在を調べたところ、それらCD163陽性のM2マクロファージはBDNFを発現していることが確認された。シュワン細胞とM2マクロファージの相互作用をさらに解析するために、ヒト歯髄組織からシュワン細胞を取り出し、in vitroにおいてTHP-1細胞由来マクロファージ(M0)との共培養を試みた。その結果、シュワン細胞に接触しとどまっているTHP-1マクロファージは、その形を球状から紡錘形に変化させ、CD163陽性のM2マクロファージに分化することが明らかとなった。歯髄は神経線維が非常に豊富な組織である。シュワン細胞はその神経線維を物理的にあるいは様々なサイトカインを分泌することで直接的に保護している。一方で、シュワン細胞は、神経線維を破壊するM1型マクロファージをM2型に変化させることで神経線維を保護していることが示唆された。

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  • The clarification of the pulp generation mechanism to improve the odontoblast differentiation by the transporter of ascorbic acid.

    Grant number:19K10147

    2019.4 - 2020.3

    System name:Grants-in-Aid for Scientific Research

    Research category:Grant-in-Aid for Scientific Research (C)

    Awarding organization:Japan Society for the Promotion of Science

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    Grant amount:\4290000 ( Direct Cost: \3300000 、 Indirect Cost:\990000 )

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  • A study on the mechanisms of induction and differentiation of stem cells during wound healing and regeneration of dentin-pulp complex

    Grant number:16H05516

    2016.4 - 2020.3

    System name:Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (B)

    Research category:Grant-in-Aid for Scientific Research (B)

    Awarding organization:Japan Society for the Promotion of Science

    YOSHIBA Kunihiko

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    Grant amount:\17030000 ( Direct Cost: \13100000 、 Indirect Cost:\3930000 )

    This study aimed to elucidate the mechanism of dental pulp wound healing and reparative dentinogenesis. We investigated the cellular events and expression of related factors after direct pulp capping or pulpotomy. The results suggested that α-SMA-positive myofibroblasts are progenitors of odontoblast-like cells and that TGF-β1 and EDA-fibronectin are involved in their differentiation. In addition, bone marrow-derived mesenchymal cells, fibrocytes, appeared transiently in the early stages and M2 phenotype macrophages colocalized with Schwann cells in healthy as well as inflamed pulp. It was suggested that various types of cells are involved in pulp wound healing and repair process.

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  • Role of alpha-SMA expressing cells in dental pulp wound healing and regeneration

    Grant number:16K11546

    2016.4 - 2019.3

    System name:Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (C)

    Research category:Grant-in-Aid for Scientific Research (C)

    Awarding organization:Japan Society for the Promotion of Science

    Yoshiba Nagako

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    Grant amount:\4680000 ( Direct Cost: \3600000 、 Indirect Cost:\1080000 )

    Dental pulp is a soft connective tissue encased in mineralized dentine. When dentine continuity is disrupted by caries or traumatic injury, the resident odontoblasts are also destroyed at that site. Pulp capping agents can help to cover the site through the production of a mineralized matrix through the activation of newly differentiated odontoblast-like cells, however, the exact mechanisms of the process have not been fully revealed. The present study has demonstrated that α-SMA expressing cells play a crucial role in the wound healing.

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  • The role of prostaglandin E2 for wound healing of dental pulp : focusing the transporters and prostaglandin E2 receptors

    Grant number:16K20450

    2016.4 - 2019.3

    System name:Grants-in-Aid for Scientific Research Grant-in-Aid for Young Scientists (B)

    Research category:Grant-in-Aid for Young Scientists (B)

    Awarding organization:Japan Society for the Promotion of Science

    Naoto Ohkura

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    Grant amount:\3770000 ( Direct Cost: \2900000 、 Indirect Cost:\870000 )

    This study attempted to analyze the mRNA expression and localization of prostaglandin transporter (PGT) and prostaglandin receptors (EPs) receptors using real-time PCR and immunohistofluorescent staining in a rat model of pulpotomy with mineral trioxide aggregate capping. Moreover, to elucidate the angiogenesis stimuli on the EPs signaling pathway, mRNA expression levels of PGT, EP2, and EP4 were analyzed in cultured human pulp tissues challenged with EPs agonists. This study provides insights into the functions of PGE2 via PGT and EPs in the healing dentin/pulp complex, angiogenesis, and neuroprotection in the rat moral pulp, whereas PGE2 via MRP4 might mediate angiogenesis induced in acute phase. Further studies may be helpful developing new therapeutic targets for dental disease.

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  • Contribution of Myofibroblasts in Dental Pulp Healing and Regeneration

    Grant number:25462952

    2013.4 - 2016.3

    System name:Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (C)

    Research category:Grant-in-Aid for Scientific Research (C)

    Awarding organization:Japan Society for the Promotion of Science

    YOSHIBA NAGAKO, YOSHIBA KUNIHIKO, OHKURA NAOTO

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    Grant amount:\4810000 ( Direct Cost: \3700000 、 Indirect Cost:\1110000 )

    Alpha-smooth muscle actin (α-SMA) is a marker of myofibroblasts, which play a central role in wound healing. Fibrillin-1 is a major constituent of microfibrils and an extracellular-regulator of TGF-β1, an important cytokine for myofibroblasts differentiation. We examined alterations of α-SMA and fibrillin-1 expression in organotypic culture of dental pulp. After 7 days of culture, most fibroblasts and odontoblasts were immunoreactive for α-SMA with a significant increase of α-SMA mRNA expression. Furthermore, immunostaining of fibrillin-1 became faint with mRNA downregulation. Administration of inhibitors for extracellular matrix proteases resulted in the recovery of fibrillin-1 immunostaining, and fibroblasts lost their immunoreactivity for α-SMA with mRNA downregulation. These findings suggest that fibrillin-1 degradation and downregulation might be implicated in the myofibroblasts differentiation in dental pulp wound healing.

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  • Membrane transport protein mediatted prostaglandin E2 transport in experimentally inflmaed rat dental pulp

    Grant number:25861794

    2013.4 - 2016.3

    System name:Grants-in-Aid for Scientific Research Grant-in-Aid for Young Scientists (B)

    Research category:Grant-in-Aid for Young Scientists (B)

    Awarding organization:Japan Society for the Promotion of Science

    OHKURA Naoto

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    Grant amount:\4030000 ( Direct Cost: \3100000 、 Indirect Cost:\930000 )

    This study attempted to analyze the mRNA and protein expression of Mrp4, Pgt and EPs receptors with real-time PCR and immunofluorescent staining in lipopolysaccharide-inflamed rat incisor pulp tissue. Moreover, the amounts of PGE2 released from the inflamed pulp tissue in the presence of absence of inhibitors were assessed by using an enzyme-linked immunosorbent assay. Mrp4, Pgt, and mPGES expression were detected in the endothelial cells of normal and LPS-inflamed rat incisor pulp tissue, suggesting that these cells are associated with the biosynthesis and transmembrane efflux transport pathway of PGE2. Moreover, EP2 and EP4 were expressed in odontoblasts and endothelial cells, suggesting that EP2 and EP4 might play various roles in dental pulp inflammation by binding the PGE2.

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  • Application of activated macrophages derived from dental pulp stem cells to dental pulp tissue engineering

    Grant number:25670808

    2013.4 - 2015.3

    System name:Grants-in-Aid for Scientific Research Grant-in-Aid for Challenging Exploratory Research

    Research category:Grant-in-Aid for Challenging Exploratory Research

    Awarding organization:Japan Society for the Promotion of Science

    OKIJI Takashi, YOSHIBA Nagako, YOSHIBA Kunihiko, KANEKO Tomoatsu, SHIGETANI Yoshimi, OHKURA Naoto

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    Grant amount:\3640000 ( Direct Cost: \2800000 、 Indirect Cost:\840000 )

    This study aimed to examine the role of macrophages in the regeneration of dental pulp tissues. After co-implantation of ED2-positive macrophages isolated from rat dental pulp with stem cells into the coronal pulp chamber of pulpotomized rat molars, expression of macrophage markers in the engineered pulp tissue was analyzed. When IL-4 treated ED2-positive macrophages were co-implanted with stem cells, the engineered tissues showed a significant increase in the number of ED2-positive cells and the expression of CD34 and CD163 mRNAs. These findings suggest that M2 macrophages show proliferation and act as wound healing macrophages after co-implantation of macrophages with stem cells.

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  • 歯髄炎症発症時におけるPGE2輸送機構の戦略的遺伝的解明

    Grant number:23792168

    2011.4 - 2012.3

    System name:科学研究費助成事業 若手研究(B)

    Research category:若手研究(B)

    Awarding organization:日本学術振興会

    大倉 直人

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    Grant amount:\4160000 ( Direct Cost: \3200000 、 Indirect Cost:\960000 )

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Teaching Experience (researchmap)

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Teaching Experience

  • 生涯にわたる歯と咬合

    2023
    Institution name:新潟大学

  • 齲蝕学

    2022
    Institution name:新潟大学

  • 保存修復学実習

    2022
    Institution name:新潟大学

  • 齲蝕学

    2020
    Institution name:新潟大学

  • 歯内療法学実習

    2017
    -
    2020
    Institution name:新潟大学