Updated on 2022/08/05

写真a

 
SAKAGUCHI Kohei
 
Organization
Nagoya University Hospital Oral and Maxillofacial Surgery Assistant Professor
Title
Assistant Professor

Degree 1

  1. 博士(医学) ( 2017.4   名古屋大学 ) 

Research Interests 6

  1. 骨再生

  2. 歯周組織再生

  3. 顎関節症

  4. 幹細胞

  5. 培養上清

  6. 幹細胞由来液性因子

Research Areas 2

  1. Life Science / Surgical dentistry

  2. Life Science / Regenerative dentistry and dental engineering

Research History 2

  1. 名古屋大学医学部附属病院   歯科口腔外科   助教

    2022.4

  2. 名古屋大学医学部附属病院   歯科口腔外科   病院助教

    2021.11 - 2022.3

Professional Memberships 9

  1. The International Association for Dental Research (IADR)

  2. 日本口腔外科学会   認定医

  3. 日本顎関節学会   認定医

  4. 日本口腔インプラント学会

  5. 日本口腔顔面痛学会

  6. 日本口腔科学会

  7. 日本化学療法学会   抗菌化学療法認定歯科医師

  8. ICD制度協議会   インフェクションコントロールドクター

  9. 日本顎変形症学会

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Awards 5

  1. 優秀ポスター発表賞(ゴールドリボン賞)

    2018.11   第63回日本口腔外科学会総会・学術大会   骨髄間葉系幹細胞が分泌するエクソソームによる新規骨再生法

    坂口晃平,酒井陽,若山有紀子,鶴田剛士,渡邉純奈,片桐渉,日比英晴

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    Award type:Award from Japanese society, conference, symposium, etc.  Country:Japan

  2. 若手優秀ポスター発表賞(共同)

    2018.5   第72回NPO法人日本口腔科学会学術集会   ビスホスホネート関連顎骨壊死に対する間葉系幹細胞由来エクソソームを用いた治療の 可能性

    渡邊純奈,酒井陽,岡部一登,若山有紀子,坂口晃平,鶴田剛士,日比英晴

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    Award type:Award from Japanese society, conference, symposium, etc.  Country:Japan

  3. 学術奨励最優秀賞(覚道健治賞)(共同)

    2017.7   日本顎関節学会   顎関節症に対する運動療法の短期的効果

    山口賀大, 佐久間重光, 遠渡将輝, 坂口晃平, 田口慧, 小林里奈, 足立充, 伊藤裕, 田口望, 日比英晴

  4. 優秀ポスター発表賞(共同)

    2014.9   第59回日本口腔外科学会総会・学術大会   乳歯幹細胞由来培養上清を用いた末梢神経再生治療の可能性

    椙村有紀子,片桐渉,大杉将嗣,河合孝真,緒方謙一,坂口晃平,日比英晴,上田実

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    Award type:Award from Japanese society, conference, symposium, etc.  Country:Japan

  5. 優秀ポスター発表賞(共同)

    2013.9   第58回日本口腔外科学会総会・学術大会   ラットBRONJ様モデルにおける幹細胞培養上清由来液性因子を用いた治療法の検討

    緒方謙一,片桐渉,大杉将嗣,河合孝真,椙村有紀子,坂口晃平,日比英晴,中村誠司,上田実

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    Award type:Award from Japanese society, conference, symposium, etc.  Country:Japan

 

Papers 11

  1. Immediate curative effects of exercise therapy in patients with myalgia of the masticatory muscles Reviewed

    Sakaguchi K, Taguchi N, Kobayashi R, Taguchi K, Okada K, Kinoshita F, Hibi H

    Journal of Oral Rehabilitation     2022.7

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

    DOI: 10.1111/joor.13355. Online ahead of print.

  2. Dental pulp stem cell-derived small extracellular vesicle in irradiation-induced senescence

    Dong Jiao, Sakai Kiyoshi, Koma Yoshiro, Watanabe Junna, Liu Kehong, Maruyama Hiroshi, Sakaguchi Kohei, Hibi Hideharu

    BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS   Vol. 575   page: 28 - 35   2021.10

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    Language:Japanese   Publisher:Biochemical and Biophysical Research Communications  

    Small extracellular vesicles (sEV) facilitate signaling molecule transfer among cells. We examined the therapeutic efficacy of human dental pulp stem cell-derived sEV (hDPSC-sEV) against cellular senescence in an irradiated-submandibular gland mouse model. Seven-week-old mice were exposed to 25 Gy radiation and randomly assigned to control, phosphate-buffered saline (PBS), or hDPSC-sEV groups. At 18 days post-irradiation, saliva production was measured; histological and reverse transcription-quantitative PCR analyses of the submandibular glands were performed. The salivary flow rate did not differ significantly between the PBS and hDPSC-sEV groups. AQP5-expressing acinar cell numbers and AQP5 expression levels in the submandibular glands were higher in the hDPSC-sEV group than in the other groups. Furthermore, compared with non-irradiated mice, mice in the 25 Gy + PBS group showed a high senescence-associated-β-galactosidase-positive cell number and upregulated senescence-related gene (p16INK4a, p19Arf, p21) and senescence-associated secretory phenotypic factor (MMP3, IL-6, PAI-1, NF-κB, and TGF-β) expression, all of which were downregulated in the hDPSC-sEV group. Superoxide dismutase levels were lower in the PBS group than in the hDPSC-sEV group. In summary, hDPSC-sEV reduced inflammatory cytokine and senescence-related gene expression and reversed oxidative stress in submandibular cells, thereby preventing irradiation-induced cellular senescence. Based on these results, we hope to contribute to the development of innovative treatment methods for salivary gland dysfunction that develops after radiotherapy for head and neck cancer.

    DOI: 10.1016/j.bbrc.2021.08.046

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  3. A case involving tooth extraction in a patient with acquired von Willebrand syndrome

    SAWAKI Tadashi, OKABE Kazuto, SAKAGUCHI Kohei, SAKAKURA Hiroki, KANEKO Ryuji, HIBI Hideharu

    Japanese Journal of Oral and Maxillofacial Surgery   Vol. 67 ( 7 ) page: 459 - 463   2021

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    Language:Japanese   Publisher:Japanese Society of Oral and Maxillofacial Surgeons  

    <p>A 42-year-old woman diagnosed with acquired von Willebrand syndrome (AvWS) was referred to our department for pain in the left mandibular third molar. vWF-containing concentrates (Confact F<sup>®</sup>) were transfused before and after tooth extraction for hemostatic management. Although initial hemostasis was achieved, persistent postoperative bleeding was observed 3 days later. Therefore, additional hemostatic management using a mouth splint with gingival packing and increased transfusion of Confact F<sup>®</sup> was required. After this, no further bleeding was observed, and the administration of Confact F<sup>®</sup> was ceased 14 days post-operation. The stitches were removed 24 days post-operation. Two years and 11 months after the operation, the clinical course is good. The pathophysiology of AvWS has not been elucidated, and an effective treatment has not yet been established. Therefore, reliable hemostatic management in cooperation with hematology is necessary.</p>

    DOI: 10.5794/jjoms.67.459

  4. Peripheral Nerve Regeneration in a Novel Rat Model of Dysphagia. Reviewed International journal

    Kiyoshi Sakai, Takeshi Tsuruta, Junna Watanabe, Yukiko Sugimura, Kohei Sakaguchi, Wataru Katagiri, Hideharu Hibi

    Methods in molecular biology (Clifton, N.J.)   Vol. 2155   page: 107 - 113   2020

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    Language:English   Publishing type:Research paper (scientific journal)  

    The superior laryngeal nerve (SLN) is known to play an essential role in the laryngeal reflex and swallowing. Damage to the SLN causes difficulty swallowing, that is, dysphagia. We successfully developed a novel rat model of dysphagia by SLN injury, in which we could evaluate the neuroregenerative capacity of stem cell from human exfoliated deciduous teeth (SHED). The dysphagic rats exhibit weight loss and altered drinking patterns. Furthermore, SLN injury induces a delayed onset of the swallowing reflex and accumulation of laryngeal debris in the pharynx. This rat model was used to evaluate the systemic application of SHED-conditioned medium (SHED-CM) as a therapeutic candidate for dysphagia. We found that SHED-CM promoted functional recovery and significant axonal regeneration in SLNs through the polarization shift of macrophages from activated inflammatory macrophages (M1) to anti-inflammatory macrophages (M2) and angiogenesis. This chapter describes the establishment of SLN-injury induced dysphagia rat model and the preparation and application of SHED-CM.

    DOI: 10.1007/978-1-0716-0655-1_9

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  5. Secretomes of mesenchymal stem cells induce early bone regeneration by accelerating migration of stem cells Reviewed

    Kenichi Ogata, Masashi Osugi, Takamasa Kawai, Yukiko Wakayama, Kohei Sakaguchi, Seiji Nakamura, Wataru Katagiri

    Journal of Oral and Maxillofacial Surgery, Medicine, and Pathology   Vol. 30 ( 5 ) page: 445 - 451   2018.9

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    Language:Japanese   Publishing type:Research paper (scientific journal)   Publisher:Elsevier Ltd  

    Objective: We previously reported that secretomes from human bone marrow-derived mesenchymal stem cells (MSC-CM) have a strong potential to accelerate bone regeneration. The most important initial step for bone regeneration is osteoprogenitor cell migration to bone defects. We hypothesized that MSC: CM enhance the migration of endogenous stem cells earlier to the local lesioned part. In this study
    we investigated the potential of MSCCM to induce in vivo early bone regeneration by accelerating cell migration in a rat calvarial bone defect model. Materials and methods: Cytokine array analysis was performed to assess the types of cytokines included in MSC-CM. Bone defects (5 mm in diameter) were created in the calvarial bones of rats, and the damaged areas were implanted with atelocollagen suspended in MSC-CM or phosphate buffered saline. After 2 and 4 weeks, radiographic and histological analyses were performed. Furthermore, rat mesenchymal stem cells (rMSCs) were labeled with the lipophilic tracer 1,1-dioctadecyl-3,3,3,3-tetramethylindotricarbocyanine iodide (DiR), and the rats were photographed at various times after injection of the DiR-labeled rMSCs using in vivo imaging. Results: MSC-CM contained many factors with respect to cell migration and tissue regeneration. Bone regeneration in rat calvaria was observed earliest in the MSC-CM implantation group. Migration of the labeled rMSCs from the tail toward the calvaria, where MSC-CM was implanted, was observed during the first 24 h after injection in the MSC-CM implantation group using in vivo imaging. Immunohistochemistry also indicated early cell migration. Conclusion: MSC-CM enhanced the migration of endogenous stem cells facilitating earlier bone regeneration.

    DOI: 10.1016/j.ajoms.2018.04.002

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  6. Angiogenesis in newly regenerated bone by secretomes of human mesenchymal stem cells. Reviewed International journal

    Wataru Katagiri, Takamasa Kawai, Masashi Osugi, Yukiko Sugimura-Wakayama, Kohei Sakaguchi, Taku Kojima, Tadaharu Kobayashi

    Maxillofacial plastic and reconstructive surgery   Vol. 39 ( 1 ) page: 8 - 8   2017.12

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    Language:Japanese   Publishing type:Research paper (scientific journal)  

    BACKGROUND: For an effective bone graft for reconstruction of the maxillofacial region, an adequate vascular network will be required to supply blood, osteoprogenitor cells, and growth factors. We previously reported that the secretomes of bone marrow-derived mesenchymal stem cells (MSC-CM) contain numerous growth factors such as insulin-like growth factor (IGF)-1, transforming growth factor (TGF)-β1, and vascular endothelial growth factor (VEGF), which can affect the cellular characteristics and behavior of regenerating bone cells. We hypothesized that angiogenesis is an important step for bone regeneration, and VEGF is one of the crucial factors in MSC-CM that would enhance its osteogenic potential. In the present study, we focused on VEGF in MSC-CM and evaluated the angiogenic and osteogenic potentials of MSC-CM for bone regeneration. METHODS: Cytokines in MSC-CM were measured by enzyme-linked immunosorbent assay (ELISA). Human umbilical vein endothelial cells (HUVECs) were cultured with MSC-CM or MSC-CM with anti-VEGF antibody (MSC-CM + anti-VEGF) for neutralization, and tube formation was evaluated. For the evaluation of bone and blood vessel formation with micro-computed tomography (micro-CT) and for the histological and immunohistochemical analyses, a rat calvarial bone defect model was used. RESULTS: The concentrations of IGF-1, VEGF, and TGF-β1 in MSC-CM were 1515.6 ± 211.8 pg/mL, 465.8 ± 108.8 pg/mL, and 339.8 ± 14.4 pg/mL, respectively. Tube formation of HUVECs, bone formation, and blood vessel formation were increased in the MSC-CM group but decreased in the MSC-CM + anti-VEGF group. Histological findings suggested that new bone formation in the entire defect was observed in the MSC-CM group although it was decreased in the MSC-CM + anti-VEGF group. Immunohistochemistry indicated that angiogenesis and migration of endogenous stem cells were much more abundant in the MSC-CM group than in the MSC-CM + anti-VEGF group. CONCLUSIONS: VEGF is considered a crucial factor in MSC-CM, and MSC-CM is proposed to be an adequate therapeutic agent for bone regeneration with angiogenesis.

    DOI: 10.1186/s40902-017-0106-4

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  7. Clinical Study of Bone Regeneration by Conditioned Medium From Mesenchymal Stem Cells After Maxillary Sinus Floor Elevation. Reviewed International journal

    Wataru Katagiri, Junna Watanabe, Naoto Toyama, Masashi Osugi, Kohei Sakaguchi, Hideharu Hibi

    Implant dentistry   Vol. 26 ( 4 ) page: 607 - 612   2017.8

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    Language:Japanese   Publishing type:Research paper (scientific journal)   Publisher:LIPPINCOTT WILLIAMS & WILKINS  

    OBJECTIVE: This clinical study was undertaken to evaluate the safety of use of the secretome of bone marrow-derived mesenchymal stem cells (MSC-CM) for maxillary sinus floor elevation (SFE). MATERIALS AND METHODS: MSC-CM was prepared from conditioned medium from human bone marrow-derived MSCs. Six partially edentulous patients were enrolled in the study. MSC-CM was mixed with porous beta-tricalcium phosphate (β-TCP) and implanted in 4 patients (experimental group), whereas only β-TCP was implanted in the other 2 patients (control group). Six months after SFE, bone biopsies and histological assessments were performed. RESULTS: Bone formation was clinically confirmed in all cases. Although Hounsfield units in computed tomography images were not significantly different between the groups, histological analysis revealed a significant difference in newly formed bone area between the groups. In particular, bone volume in the center of the augmented area was significantly greater in the MSC-CM group. Newly formed bone consisted of lamellar bone in the MSC-CM group but woven bone in the β-TCP group. CONCLUSION: The secretome of bone marrow-derived mesenchymal stem cells (MSC-CM) was used safely and has great osteogenic potential for regenerative medicine of bone.

    DOI: 10.1097/ID.0000000000000618

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  8. A defined mix of cytokines mimics conditioned medium from cultures of bone marrow-derived mesenchymal stem cells and elicits bone regeneration. Reviewed International journal

    Wataru Katagiri, Kohei Sakaguchi, Takamasa Kawai, Yukiko Wakayama, Masashi Osugi, Hideharu Hibi

    Cell proliferation   Vol. 50 ( 3 )   2017.6

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    Language:Japanese   Publishing type:Research paper (scientific journal)   Publisher:WILEY  

    OBJECTIVES: We previously reported that conditioned medium from cultures of bone marrow-derived mesenchymal stem cells have strong potential to accelerate bone regeneration. We now examine in vitro and in vivo a defined cytokine cocktail that mimics the effects of conditioned medium on bone regeneration. MATERIALS AND METHODS: A cocktail of recombinant human insulin-like growth factor-1, vascular endothelial growth factor-A and transforming growth factor-β1 was prepared at concentrations similar to those in conditioned medium. Conversely, these cytokines were depleted from conditioned medium, and the effects of the cocktail, the conditioned medium and the cytokine-depleted conditioned medium on bone regeneration were evaluated in vitro and in vivo. RESULTS: The cytokine cocktail and conditioned medium enhanced cell migration, tube formation, and expression of osteogenic and angiogenic genes. Depletion of cytokines significantly decreased the effects of conditioned medium in vitro. Similarly, the cytokine cocktail and conditioned medium, but not cytokine-depleted medium, increased bone regeneration in damaged rat calvarial bone. Immunohistochemistry indicated that the cytokine cocktail and conditioned medium strongly enhanced recruitment of endogenous stem cells and endothelial cells. CONCLUSIONS: The data indicate that the cytokine cocktail and conditioned medium enhance the migration of stem cells and endothelial cells to damaged bone, and elicit osteogenesis and angiogenesis.

    DOI: 10.1111/cpr.12333

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  9. Periodontal tissue regeneration using the cytokine cocktail mimicking secretomes in the conditioned media from human mesenchymal stem cells. Reviewed International journal

    Kohei Sakaguchi, Wataru Katagiri, Masashi Osugi, Takamasa Kawai, Yukiko Sugimura-Wakayama, Hideharu Hibi

    Biochemical and biophysical research communications   Vol. 484 ( 1 ) page: 100 - 106   2017.2

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    Language:Japanese   Publishing type:Research paper (scientific journal)   Publisher:ACADEMIC PRESS INC ELSEVIER SCIENCE  

    Secretomes in the conditioned media from human mesenchymal stem cells (MSC-CM) were previously demonstrated to promote periodontal tissue regeneration. By mixing insulin-like growth factor-1, vascular endothelial growth factor-A, and transforming growth factor-β1 which were included in MSC-CM, we made the cytokine cocktail (CC) mimicking MSC-CM, and then evaluated its efficacy on periodontal tissue regeneration. In vitro, CC promoted the migration of dog bone marrow-derived stem cells and periodontal ligament cells, and the tube formation of human umbilical vein endothelial cells. In vivo, class II furcation defects were surgically created at premolars in dogs. After 4 weeks of vinylpolysiloxane-induced inflammation, defects were filled with or without CC mixed in hydroxypropyl cellulose, or enamel matrix derivative (EMD). After 8 weeks, periodontal tissues were evaluated histologically and immunohistochemically. CC showed promotional effects on angiogenesis and formation of new bone and cementum. Osteogenesis by CC was greater than that by EMD and cementogenesis by CC was as well as that by EMD. CC may be promising for periodontal tissue regeneration.

    DOI: 10.1016/j.bbrc.2017.01.065

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  10. Short-term effectiveness of exercise therapy for temporomandibular disorders:-Preliminary study for temporomandibular joint anterior disc displacement without reduction-

    YAMAGUCHI Yoshihiro, HIBI Hideharu, SAKUMA Shigemitsu, ENDO Masaki, SAKAGUCHI Kohei, TAGUCHI Kei, KOBAYASHI Rina, ADACHI Mitsuru, ITO Hiroshi, TAGUCHI Nozomu

    Journal of the Japanese Society for the Temporomandibular Joint   Vol. 28 ( 2 ) page: 126 - 134   2016

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    Language:Japanese   Publisher:The Japanese Society for Temporomandibular Joint  

    <p>Therapeutic exercise increases range of motion, decreases pain, and shortens the duration of symptoms. However, few studies have assessed the effectiveness of therapeutic exercise by using quantitative methods. In this study, we prescribed a range of temporomandibular joint motion exercises involving both passive exercises performed by a dentist and self-traction therapy performed by the patient, and evaluated the short-term effectiveness of the treatment in patients with temporomandibular joint anterior disc displacement without reduction.</p><p>We enrolled 45 patients who had moderate or severe TMJ function. We asked them to perform therapeutic exercises and evaluated their clinical symptoms (maximum opening distance, pain at rest, pain on motion, pain on mastication, and degree of difficulty in performing activities of daily living) at the first visit and at the first follow-up visit approximately 2 weeks later.</p><p>The results demonstrated statistically significant improvements in maximum opening distance, pain on motion, pain on mastication and degree of difficulty in performing activities of daily living (p<0.001). The positive outcomes were considered the result of improvements in range of joint motion and expansion of the joint cavity, resulting in smoother movement of the condylar translation. The results of this study suggest that our therapeutic exercise program is a potentially beneficial conservative treatment for the short-term reduction of symptoms accompanying temporomandibular joint anterior disc displacement without reduction.</p>

    DOI: 10.11246/gakukansetsu.28.126

  11. Peripheral Nerve Regeneration by Secretomes of Stem Cells from Human Exfoliated Deciduous Teeth. Reviewed International journal

    Yukiko Sugimura-Wakayama, Wataru Katagiri, Masashi Osugi, Takamasa Kawai, Kenichi Ogata, Kohei Sakaguchi, Hideharu Hibi

    Stem cells and development   Vol. 24 ( 22 ) page: 2687 - 2699   2015.11

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    Language:Japanese   Publishing type:Research paper (scientific journal)   Publisher:MARY ANN LIEBERT, INC  

    Peripheral nerve regeneration across nerve gaps is often suboptimal, with poor functional recovery. Stem cell transplantation-based regenerative therapy is a promising approach for axon regeneration and functional recovery of peripheral nerve injury; however, the mechanisms remain controversial and unclear. Recent studies suggest that transplanted stem cells promote tissue regeneration through a paracrine mechanism. We investigated the effects of conditioned media derived from stem cells from human exfoliated deciduous teeth (SHED-CM) on peripheral nerve regeneration. In vitro, SHED-CM-treated Schwann cells exhibited significantly increased proliferation, migration, and the expression of neuron-, extracellular matrix (ECM)-, and angiogenesis-related genes. SHED-CM stimulated neuritogenesis of dorsal root ganglia and increased cell viability. Similarly, SHED-CM enhanced tube formation in an angiogenesis assay. In vivo, a 10-mm rat sciatic nerve gap model was bridged by silicon conduits containing SHED-CM or serum-free Dulbecco's modified Eagle's medium. Light and electron microscopy confirmed that the number of myelinated axons and axon-to-fiber ratio (G-ratio) were significantly higher in the SHED-CM group at 12 weeks after nerve transection surgery. The sciatic functional index (SFI) and gastrocnemius (target muscle) wet weight ratio demonstrated functional recovery. Increased compound muscle action potentials and increased SFI in the SHED-CM group suggested sciatic nerve reinnervation of the target muscle and improved functional recovery. We also observed reduced muscle atrophy in the SHED-CM group. Thus, SHEDs may secrete various trophic factors that enhance peripheral nerve regeneration through multiple mechanisms. SHED-CM may therefore provide a novel therapy that creates a more desirable extracellular microenvironment for peripheral nerve regeneration.

    DOI: 10.1089/scd.2015.0104

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Books 1

  1. Peripheral Nerve Regeneration in a Novel Rat Model of Dysphagia

    Kiyoshi Sakai , Takeshi Tsuruta, Junna Watanabe, Yukiko Sugimura, Kohei Sakaguchi, Wataru Katagiri, Hideharu Hibi( Role: Joint author)

    2020.5 

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    Total pages:7   Language:English

MISC 42

  1. シェーグレン症候群の唾液腺細胞老化に幹細胞由来細胞外小胞が与える影響

    小間 義朗, 酒井 陽, 渡邊 純奈, 椙村 有紀子, 坂口 晃平, 丸山 裕, 日比 英晴

    日本口腔科学会雑誌   Vol. 70 ( 2 ) page: 141 - 141   2021.7

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    Language:Japanese   Publisher:(NPO)日本口腔科学会  

  2. 後天性von Willebrand症候群患者の抜歯経験

    澤木 廉, 岡部 一登, 坂口 晃平, 坂倉 寛紀, 兼子 隆次, 日比 英晴

    日本口腔外科学会雑誌   Vol. 67 ( 7 ) page: 459 - 463   2021.7

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    Language:Japanese   Publisher:(公社)日本口腔外科学会  

    42歳女性。左下顎歯8の周囲歯肉痛を主訴とした。歯冠周囲炎と診断のもと抜歯術が計画されたが、本症例はIgG-λ型単クローン性ガンマグロブリン血症を契機に発症した後天性von Willebrand症候群の既往があり、抜歯術開始1時間前にコンファクトFを3000単位投与した。局所麻酔薬には酒石酸水素エピネフリン含有2%塩酸リドカインを使用し、歯冠を分割して左下顎歯8を摘出した。術後もコンファクトFの投与を継続した結果、異常出血や知覚鈍麻、治癒遅延はなく、術後18日目に退院となった。

  3. 汎発性帯状疱疹を併発した不完全型Ramsay-Hunt症候群の1例

    宮坂 紗季, 岡部 一登, 酒井 陽, 若山 有紀子, 大森 正裕, 坂口 晃平, 日比 英晴

    日本口腔科学会雑誌   Vol. 70 ( 1 ) page: 71 - 71   2021.3

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    Language:Japanese   Publisher:(NPO)日本口腔科学会  

  4. 骨造成後の骨補填材に感染が波及した1例

    渡邊 純奈, 岡部 一登, 酒井 陽, 藤尾 正人, 椙村 有紀子, 坂口 晃平, 佐世 暁, 日比 英晴

    日本口腔インプラント学会誌   Vol. 33 ( 特別号 ) page: 293 - 293   2020.9

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    Language:Japanese   Publisher:(公社)日本口腔インプラント学会  

  5. 骨再生についてのわれわれの取り組みの変遷

    岡部 一登, 藤尾 正人, 酒井 陽, 椙村 有紀子, 坂口 晃平, 渡邊 純奈, 日比 英晴

    日本口腔インプラント学会誌   Vol. 33 ( 特別号 ) page: 410 - 410   2020.9

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    Language:Japanese   Publisher:(公社)日本口腔インプラント学会  

  6. 咀嚼筋痛障害に対する運動療法の即時的治療効果の検証

    坂口 晃平, 田口 望, 小林 里奈, 田口 慧, 岡田 啓, 日比 英晴

    日本顎関節学会雑誌   Vol. 32 ( Suppl. ) page: 76 - 76   2020.7

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    Language:Japanese   Publisher:(一社)日本顎関節学会  

  7. 低血圧麻酔下の顎矯正手術中に心停止をきたした顎変形症の一例

    坂口 晃平, 藤尾 正人, 酒井 陽, 日比 英晴

    日本顎変形症学会雑誌   Vol. 30 ( 2 ) page: 172 - 172   2020.5

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    Language:Japanese   Publisher:(NPO)日本顎変形症学会  

  8. シリコーンアレルギー患者における周術期管理の経験

    坂口 晃平, 岡部 一登, 佐藤 康太郎, 澤木 廉, 日比 英晴

    有病者歯科医療   Vol. 29 ( 1 ) page: 148 - 148   2020.4

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    Language:Japanese   Publisher:(一社)日本有病者歯科医療学会  

  9. 歯冠修復後に進行した歯髄炎による対顎歯への関連痛の1例

    加納 史也, 西須 大徳, 神尾 尚伸, 鶴田 剛士, 坂口 晃平, 日比 英晴, 山本 朗仁

    日本口腔診断学会雑誌   Vol. 33 ( 1 ) page: 90 - 91   2020.2

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  10. 後天性von Willebrand症候群患者における抜歯経験

    赤坂 廉, 岡部 一登, 坂口 晃平, 日比 英晴

    有病者歯科医療   Vol. 28 ( 6 ) page: 410 - 410   2019.12

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  11. 歯冠修復後に進行した歯髄炎による対顎歯への関連痛の1例

    加納 史也, 西須 大徳, 神尾 尚伸, 鶴田 剛士, 坂口 晃平, 日比 英晴, 山本 朗仁

    日本口腔内科学会雑誌   Vol. 25 ( 2 ) page: 80 - 81   2019.12

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  12. 後天性von Willebrand症候群患者における抜歯経験

    赤坂廉, 岡部一登, 坂口晃平, 日比英晴

    歯科薬物療法   Vol. 38 ( 2 ) page: 105 - 105   2019.3

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  13. 運動療法により即時的治療効果を認めた咀嚼筋痛障害の一例

    坂口 晃平, 小林 里奈, 田口 慧, 日比 英晴, 田口 望

    日本歯科心身医学会雑誌   Vol. 33 ( 2 ) page: 133 - 133   2018.12

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  14. 運動療法により即時的治療効果を認めた咀嚼筋痛障害の一例

    坂口 晃平, 小林 里奈, 田口 慧, 日比 英晴, 田口 望

    日本歯科心身医学会雑誌   Vol. 33 ( 2 ) page: 133 - 133   2018.12

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  15. 運動療法により即時的治療効果を認めた咀嚼筋痛障害の一例

    坂口晃平, 坂口晃平, 小林里奈, 田口慧, 日比英晴, 田口望, 田口望

    日本顎関節学会雑誌   Vol. 30   page: 145   2018.7

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  16. ビスホスホネート関連顎骨壊死に対する間葉系幹細胞由来エクソソームを用いた治療の可能性

    渡邊 純奈, 酒井 陽, 岡部 一登, 若山 有紀子, 坂口 晃平, 鶴田 剛士, 日比 英晴

    日本口腔科学会雑誌   Vol. 67 ( 2 ) page: 162 - 162   2018.7

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  17. 骨髄間葉系幹細胞由来エクソソームは骨芽細胞分化を促進する

    竹内 涼子, 片桐 渉, 遠藤 諭, 坂口 晃平, 小林 正治

    日本口腔科学会雑誌   Vol. 67 ( 2 ) page: 100 - 100   2018.7

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  18. 運動療法により即時的治療効果を認めた咀嚼筋痛障害の一例

    坂口 晃平, 小林 里奈, 田口 慧, 日比 英晴, 田口 望

    日本顎関節学会雑誌   Vol. 30 ( Suppl. ) page: 145 - 145   2018.7

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  19. 他家骨髄由来間葉系細胞の移植による骨再生の検討

    渡邊 純奈, 片桐 渉, 大杉 将嗣, 酒井 陽, 岡部 一登, 椙村 有紀子, 坂口 晃平, 鶴田 剛士, 日比 英晴

    日本口腔インプラント学会誌   Vol. 31 ( 2 ) page: E75 - E75   2018.6

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  20. ビスホスホネート関連顎骨壊死に対する間葉系幹細胞由来エクソソームを用いた治療の可能性

    渡邊純奈, 酒井陽, 岡部一登, 若山有紀子, 坂口晃平, 鶴田剛士, 日比英晴

    日本口腔科学会雑誌(Web)   Vol. 67 ( 2 ) page: 162(J‐STAGE)   2018

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  21. 骨髄由来間葉系幹細胞が分泌するエクソソームによる新たな骨再生

    坂口晃平, 酒井陽, 片桐渉, 片桐渉, 若山有紀子, 鶴田剛士, 渡邉純奈, 日比英晴

    日本再生医療学会総会(Web)   Vol. 17th   page: ROMBUNNO.P‐01‐039 (WEB ONLY)   2018

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  22. 歯髄幹細胞由来エクソソームのSchwann細胞への影響

    加納史也, 市村典久, 坂口晃平, 中道瑛司, 山口聡, 坂倉寛紀

    日本再生医療学会総会(Web)   Vol. 17th   page: ROMBUNNO.P‐01‐076 (WEB ONLY)   2018

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  23. 顎関節症に対する運動療法の臨床効果に関する検討 第3報:非復位性関節円板障害に対する即時効果

    山口賀大, 佐久間重光, 遠渡将輝, 坂口晃平, 坂口晃平, 田口慧, 小林里奈, 小木信美, 栗田賢一, 田口望, 田口望, 田口望

    日本顎関節学会雑誌   Vol. 29   page: 125   2017.7

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  24. ラット末梢性嚥下障害モデルを用いた乳歯歯髄幹細胞由来成長因子の治療効果の検討

    鶴田 剛士, 片桐 渉, 大杉 将嗣, 酒井 陽, 若山 有紀子, 坂口 晃平, 渡邊 純奈, 日比 英晴

    日本口腔科学会雑誌   Vol. 66 ( 2 ) page: 207 - 208   2017.7

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  25. 顎関節症に対する運動療法の臨床効果に関する検討(第3報) 非復位性関節円板障害に対する即時効果

    山口 賀大, 佐久間 重光, 遠渡 将輝, 坂口 晃平, 田口 慧, 小林 里奈, 小木 信美, 栗田 賢一, 田口 望

    日本顎関節学会雑誌   Vol. 29 ( Suppl. ) page: 125 - 125   2017.7

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  26. ラット末梢性嚥下障害モデルの確立と乳歯歯髄幹細胞由来成長因子による治療効果の検討

    鶴田剛士, 片桐渉, 大杉将嗣, 酒井陽, 若山有紀子, 坂口晃平, 渡邊純奈, 日比英晴

    再生医療   Vol. 16   page: 381   2017.2

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  27. ラット末梢性嚥下障害モデルを用いた乳歯歯髄幹細胞由来成長因子の治療効果の検討

    鶴田剛士, 片桐渉, 大杉将嗣, 酒井陽, 若山有紀子, 坂口晃平, 渡邊純奈, 日比英晴

    日本口腔科学会学術集会プログラム・抄録集   Vol. 71st   page: 317   2017

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  28. 顎関節症に対する運動療法の短期的効果 非復位性関節円板前方転位に対する予備的検討

    山口 賀大, 佐久間 重光, 遠渡 将輝, 坂口 晃平, 田口 慧, 小林 里奈, 足立 充, 伊藤 裕, 田口 望, 日比 英晴

    日本顎関節学会雑誌   Vol. 28 ( 2 ) page: 126 - 134   2016.8

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    運動療法は、施術直後より関節可動域を増大し、疼痛を早期に軽減させ病悩期間を短縮するものの、その効果について定量的な評価を行った研究は少ない。本研究では、術者が行う顎関節可動化療法と患者が行う自己牽引療法を1つの運動療法プログラムとして捉え、非復位性関節円板前方転位症例に実施した際の短期的治療効果を検討した。顎関節機能に中等度以上の障害が認められた45例を対象として運動療法を施行し、初診時とその約2週間後の初回再来時における臨床症状(最大開口域、安静時痛、開閉口時痛、咀嚼時痛および日常生活支障度)について評価した。その結果、最大開口域、開閉口時痛、咀嚼時痛および日常生活支障度において有意な改善を認めた(p&lt;0.001)。これら症状の改善は、運動療法により関節可動域が改善され、関節腔が拡大されることで下顎頭の動きが改善したものと考える。したがって、本運動療法プログラムは、非復位性関節円板前方転位に伴う諸症状を短期間に軽減させる有効な保存療法になる可能性が示唆された。(著者抄録)

    Other Link: https://search.jamas.or.jp/index.php?module=Default&action=Link&pub_year=2016&ichushi_jid=J02499&link_issn=&doc_id=20161005240002&doc_link_id=10.11246%2Fgakukansetsu.28.126&url=https%3A%2F%2Fdoi.org%2F10.11246%2Fgakukansetsu.28.126&type=J-STAGE&icon=https%3A%2F%2Fjk04.jamas.or.jp%2Ficon%2F00007_3.gif

  29. イヌ歯周病モデルを用いた成長因子混合剤による歯周組織再生能の検討

    坂口晃平, 片桐渉, 大杉将嗣, 若山有紀子, 鶴田剛士, 日比英晴

    再生医療   Vol. 15   page: 230   2016.2

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  30. 骨髄由来間葉系幹細胞培養上清由来液性因子を用いた新たな骨再生治療に関する臨床的検討

    外山直人, 片桐渉, 大杉将嗣, 安藤さやか, 若山有紀子, 坂口晃平, 鶴田剛士, 渡邊純奈, 日比英晴

    再生医療   Vol. 15   page: 207   2016.2

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  31. 骨髄間葉系幹細胞培養上清を模した成長因子混合剤による新規骨再生法

    坂口晃平, 片桐渉, 大杉将嗣, 河合孝真, 日比英晴

    日本口腔インプラント学会誌   Vol. 28 ( 4 ) page: 532 - 533   2015.12

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  32. 骨髄間葉系幹細胞培養上清を模した成長因子混合剤による新規骨再生法

    坂口 晃平, 片桐 渉, 大杉 将嗣, 河合 孝真, 日比 英晴

    日本口腔インプラント学会誌   Vol. 28 ( 4 ) page: 532 - 533   2015.12

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  33. 顎関節症に対する運動療法の臨床効果に関する検討 第2報:非復位性顎関節円板障害に対する短期的効果

    山口賀大, 佐久間重光, 遠渡将輝, 小菅哲, 坂口晃平, 田口慧, 森仁美, 小林里奈, 足立充, 伊藤裕, 田口望, 田口望, 田口望

    日本顎関節学会雑誌   Vol. 27   page: 154   2015.7

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  34. 顎関節症に対する運動療法の臨床効果に関する検討(第2報) 非復位性顎関節円板障害に対する短期的効果

    山口 賀大, 佐久間 重光, 遠渡 将輝, 小菅 哲, 坂口 晃平, 田口 慧, 森 仁美, 小林 里奈, 足立 充, 伊藤 裕, 田口 望

    日本顎関節学会雑誌   Vol. 27 ( Suppl. ) page: 154 - 154   2015.7

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  35. ラットBRONJ様モデルにおける幹細胞培養上清による治療効果の検討

    緒方謙一, 片桐渉, 大杉将嗣, 河合孝真, 椙村有紀子, 坂口晃平, 日比英晴, 中村誠司, 上田実

    再生医療   Vol. 14   page: 310   2015.2

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  36. 末梢神経損傷に対する乳歯歯髄幹細胞由来培養上清局所投与の有効性の検討

    椙村有紀子, 片桐渉, 大杉将嗣, 河合孝真, 緒方謙一, 坂口晃平

    再生医療   Vol. 14   page: 233   2015.2

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  37. 成長因子混合剤による新たな骨再生法

    坂口晃平, 片桐渉, 大杉将嗣, 河合孝真, 椙村友紀子, 緒方謙一, 日比英晴

    再生医療   Vol. 14   page: 273   2015.2

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  38. 顎関節症に対する運動療法の臨床効果に関する検討 第1報:復位性関節円板障害に対する短期的効果

    遠渡将輝, 佐久間重光, 小菅哲, 坂口晃平, 山口賀大, 小林里奈, 森仁美, 安藤清文, 伊藤裕, 田口望

    日本顎関節学会雑誌   Vol. 26   page: 128   2014.7

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  39. 顎関節症に対する運動療法の臨床効果に関する検討(第1報) 復位性関節円板障害に対する短期的効果

    遠渡 将輝, 佐久間 重光, 小菅 哲, 坂口 晃平, 山口 賀大, 小林 里奈, 森 仁美, 安藤 清文, 伊藤 裕, 田口 望

    日本顎関節学会雑誌   Vol. 26 ( Suppl. ) page: 128 - 128   2014.7

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  40. ラットBRONJ様モデルにおける幹細胞培養上清由来液性因子を用いた治療法の検討

    緒方謙一, 片桐渉, 大杉将嗣, 河合孝真, 椙村有紀子, 坂口晃平, 日比英晴, 中村誠司, 上田実

    再生医療   Vol. 13   page: 234   2014.1

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  41. 複数のサイトカインを組み合わせた新規骨再生法

    坂口晃平, 片桐渉, 大杉将嗣, 河合孝真, 椙村有紀子, 緒方謙一, 日比英晴, 上田実

    再生医療   Vol. 13   page: 235   2014.1

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  42. 脱落乳歯歯髄幹細胞由来液性因子を用いた末梢神経再生治療の可能性

    椙村有紀子, 片桐渉, 大杉将嗣, 河合孝真, 緒方謙一, 坂口晃平, 日比英晴, 上田実

    再生医療   Vol. 13   page: 333   2014.1

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Presentations 22

  1. Exercise therapy in patients with degenerative disease of the temporomandibular joint International conference

    Kohei Sakaguchi, Kiyoshi Sakai, Hiroshi Maruyama, Nanami Takano, Miku Imura, Nozomu Taguchi, Hideharu Hibi

    2022.6.23  The International Association for Dental Research

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    Event date: 2022.6

    Language:English   Presentation type:Oral presentation (general)  

    Country:China  

  2. Near Infrared Phot-Antimicrobial Targeting Therapy for Periodontal Disease International coauthorship International conference

    Hiroshi Maruyama, Kiyoshi Sakai, Kazuhide Sato, Kohei Sakaguchi, Yoshiro Koma, Junna Watanabe, Dong Jiao, Liu Kehong, Hideharu Hibi

    100th General Session and exhibition of the IADR  2022.6.23  The International Association for Dental Research

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    Event date: 2022.6

    Language:English   Presentation type:Oral presentation (general)  

    Country:China  

  3. 顎関節症患者に対するホームケアとしての自己牽引療法の有効性の検証

    髙野 菜名実、坂口 晃平、加藤 みなみ、丸山 裕、 井村 美孔、 酒井 陽、小林 里奈、田口 慧、田口 望、日比 英晴

    第35回一般社団法人日本顎関節学会学術大会  2022.7.2  日本顎関節学会

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    Event date: 2022.7

    Venue:北海道   Country:Japan  

  4. Exosomes derived from mesenchymal stem cells stimulated by hypoxia accelerated osteogenesis International conference

    Kohei Sakaguchi, Kiyoshi Sakai, Yukiko Sugimura-Wakayama, Yoshiro Koma, Junna Watanabe, Hiroshi Maruyama, Liu, Kehong, Dong Jiao, Hideharu Hibi

    99th General Session and exhibition of the IADR  2021.7  The International Association for Dental Research

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    Event date: 2021.7

    Language:English   Presentation type:Poster presentation  

    Venue:Boston   Country:United States  

  5. 非復位性顎関節円板障害に対する運動療法の有効性の検証とロック解除を困難にする要因の探索

    坂口晃平,田口望,小林里奈,田口慧,日比英晴

    第34回日本顎関節学会  2021.10  日本顎関節学会

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    Event date: 2021.10 - 2021.11

    Language:Japanese   Presentation type:Poster presentation  

    Venue:ウェビナー   Country:Japan  

  6. シェーグレン症候群の唾液腺細胞老化に幹細胞由来細胞外小胞が与える影響

    小間義朗,酒井陽,渡邊純奈,椙村有紀子,坂口晃平,丸山裕,日比英晴

    第75回日本口腔科学会  2021.5.13  日本口腔科学会

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    Event date: 2021.5

    Language:Japanese   Presentation type:Poster presentation  

    Venue:大阪   Country:Japan  

  7. Bone regeneration using exosomes derived from hMSCs stimulated by hypoxia International conference

    Kohei Sakaguchi, Kiyoshi Sakai, Yukiko Sugimura-Wakayama, Junna Watanabe, Hideharu Hibi.

    European Association for Osseointegration 28th Annual Scientific Meeting  2019.9  European Association for Osseointegration

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    Event date: 2019.9

    Language:English   Presentation type:Poster presentation  

    Venue:Lisbon   Country:Portugal  

  8. Effects of Mesenchymal Stem Cell-derived Exosomes for Rat BRONJ Model. International conference

    Junna Watanabe, Kiyoshi Sakai, Kazuto Okabe, Yukiko Sugimura, Kohei Sakaguchi, Takeshi Tsuruta, Hideharu Hibi.

    96th General Session and exhibition of the IADR  2018.7  The International Association for Dental Research

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    Event date: 2018.7

    Language:English   Presentation type:Poster presentation  

    Venue:London   Country:United Kingdom  

  9. Exosomes derived from mesenchymal stem cells enhanced bone regeneration. International conference

    Kohei Sakaguchi, Kiyoshi Sakai, Wataru Katagiri, Yukiko Sugimura-Wakayama, Takeshi Tsuruta, Junna Watanabe, Hideharu Hibi.

    96th General Session and exhibition of the IADR  2018.7  The International Association for Dental Research

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    Event date: 2018.7

    Language:English   Presentation type:Poster presentation  

    Venue:London   Country:United Kingdom  

  10. Conditioned media from SHED enhanced peripheral nerve regeneration. International conference

    Yukiko Sugimura, Kiyoshi Sakai, Hirotaka Wakayama, Kohei Sakaguchi, Takeshi Tsuruta, Junna Watanabe, Wataru Katagiri, Hideharu Hibi.

    96th General Session and exhibition of the IADR  2018.7  The International Association for Dental Research

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    Event date: 2018.7

    Language:English   Presentation type:Poster presentation  

    Venue:London   Country:United Kingdom  

  11. Conditioned media from bone marrow derived mesenchymal stem cells enhances bone regeneration in rat calvarial bone defects. International conference

    Masashi Osugi, Wataru Katagiri, Kohei Sakaguchi, Takamasa Kawai, Hideharu Hibi.

    31st Annual meeting of Academy of Osseointegration  2016.2  Academy of Osseointegration

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    Event date: 2016.2

    Language:English   Presentation type:Poster presentation  

    Venue:San Diego   Country:United States  

  12. Clinical Study for Bone Regeneration after Maxillary Sinus Floor Elevation by Secretomes from Mesenchymal Stem Cells. International conference

    Wataru Katagiri, Masashi Osugi, Kohei Sakaguchi, Takamasa Kawai, Hideharu Hibi.

    31st Annual meeting of Academy of Osseointegration  2016.2  Academy of Osseointegration

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    Event date: 2016.2

    Language:English   Presentation type:Poster presentation  

    Venue:San Diego   Country:United States  

  13. Novel bone regenerative medicine using the secretomes from bone marrow-derived mesenchymal stem cells. International conference

    Wataru Katagiri, Masashi Osugi, Takamasa Kawai, Yukiko Sugimura, Kennichi Ogata, Kohei Sakaguchi, Hideharu Hibi.

    93rd General Session and exhibition of the IADR  2015.3 

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    Event date: 2015.3

    Language:English   Presentation type:Poster presentation  

    Country:United States  

  14. Conditioned media from SHED enhanced peripheral nerve regeneration. International conference

    Yukiko Sugimura, Wataru Katagiri, Masashi Osugi, Takamasa Kawai, Kennichi Ogata, Kohei Sakaguchi, Hideharu Hibi.

    93rd General Session and exhibition of the IADR  2015.3  The International Association for Dental Research

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    Event date: 2015.3

    Language:English  

    Country:United States  

  15. Novel strategy of bone regeneration with the cocktail of recombinant cytokines mimicking secretomes derived from mesenchymal stem cells. International conference

    Kohei Sakaguchi, Wataru Katagiri, Masashi Osugi, Takamasa Kawai, Yukiko Sugimura, Kennichi Ogata, Hideharu Hibi, Minoru Ueda.

    European Association for Osseointegration 23rd Annual Scientific Meeting  2014.9.25  European Association for Osseointegration

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    Event date: 2014.9

    Language:English   Presentation type:Poster presentation  

    Country:Italy  

  16. Evaluation of the therapeutic effects of conditioned media from mesenchymal stem cells in rats’ bisphosphonate-related osteonecrosis of the jaw model. International conference

    Kennichi Ogata, Wataru Katagiri, Masashi Osugi, Takamasa Kawai, Yukiko Sugimura, Kohei Sakaguchi, Hideharu Hibi, Minoru Ueda.

    European Association for Osseointegration 23rd Annual Scientific Meeting  2014.9.25  European Association for Osseointegration

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    Event date: 2014.9

    Language:English   Presentation type:Poster presentation  

    Country:Italy  

  17. Conditioned media from Stem cells from human exfoliated deciduous teeth (SHED) enhanced peripheral nerve regeneration. International conference

    Yukiko Sugimura, Wataru Katagiri, Masashi Osugi, Takamasa Kawai, Kennichi Ogata, Kohei Sakaguchi, Hideharu Hibi, Minoru Ueda.

    European Association for Osseointegration 23rd Annual Scientific Meeting  European Association for Osseointegration

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    Event date: 2014.9

    Language:English   Presentation type:Poster presentation  

    Country:Italy  

  18. Secretomes from bone marrow derived mesenchymal stem cells enhanced Periodontal Tissue Regeneration. International conference

    Takamasa Kawai, Wataru Katagiri, Masashi Osugi, Yukiko Sugimura, Kennichi Ogata, Kohei Sakaguchi, Hideharu Hibi, Minoru Ueda.

    European Association for Osseointegration 23rd Annual Scientific Meeting  2014.9.25  European Association for Osseointegration

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    Event date: 2014.9

    Language:English   Presentation type:Poster presentation  

    Country:Italy  

  19. 汎発性帯状疱疹を併発した不完全型Ramsay-Hunt症候群の1例

    宮坂紗季, 岡部一登, 酒井陽, 若山有紀子, 大森正裕, 坂口晃平, 日比英晴

    第63回日本口腔科学会中部地方部会 2020年10月11日  2920.10.11 

  20. 骨造成後の骨補填材に感染が波及した1例

    渡邊純奈, 岡部一登, 酒井陽, 藤尾正人, 椙村有紀子, 坂口晃平, 佐世曉, 日比英晴

    第50回日本口腔インプラント学会記念学術大会  2020.9.19 

  21. 骨再生についてのわれわれの取り組みの変遷

    岡部一登, 藤尾正人, 酒井陽, 椙村有紀子, 坂口晃平, 渡邊純奈, 日比英晴

    第50回日本口腔インプラント学会記念学術大会  2020.9.19 

  22. シリコーンアレルギー患者における周術期管理の経験

    坂口晃平, 岡部一登, 佐藤康太郎, 澤木廉, 日比英晴

    第29回日本有病者歯科医療学会学術大会  2020 

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KAKENHI (Grants-in-Aid for Scientific Research) 2

  1. 環境負荷に応答した幹細胞が分泌する細胞外小胞による機能的血管網構築と歯周組織再生

    2022.4 - 2026.3

    日本学術振興会  若手研究 

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    Authorship:Other 

    Grant amount:\3500000

  2. 低酸素環境で刺激された幹細胞由来エクソソームを用いた新たな骨再生

    Grant number:19K19050  2019.4 - 2022.3

    科学研究費助成事業  若手研究

    坂口 晃平

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    Authorship:Principal investigator 

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

    歯を喪失した患者が歯科インプラント治療を希望するも骨量が不足している場合、骨髄間葉系幹細胞の移植などによる骨再生治療が試みられてきた。研究代表者が所属するグループでは、移植された幹細胞が分泌する液性因子とくにエクソソーム(MSCs-Exo)が骨再生に大きな役割を有していることを明らかにした。さらに予備実験で低酸素条件下で培養回収したMSCs-Exoは通常条件と比し骨再生能を増強する傾向が観察された。
    本研究では複数の低酸素条件下で培養回収したMSCs-Exoをラット頭蓋骨骨欠損モデルへ投与し低酸素条件が及ぼす影響を検証し、組織再生効果が高まる理想的な低酸素条件を明らかにし骨再生に有用なmiRNAを同定する。
    ①幹細胞から分泌されるエクソソームへ低酸素があたえる影響を評価した。21%と1%の酸素濃度で培養したMSCsからエクソソームを回収したところ、1%で培養した場合の方が有意に回収されるタンパク質量(エクソソーム量)が多かった。透過型電子顕微鏡を用いてそれらを観察したところ、1%で培養した場合の方が、直径100nmほどの大きな球体としてエクソソームが観察された。またナノトラッキングアッセイを行ったところ、1%条件の方が大きな粒径にピークをむかえる分布を示した。これらの結果から、低酸素で培養した場合、大きな粒径の小胞が分泌されるようになり、分泌量も増加することが明らかとなった。
    ②低酸素条件のMSCsから回収したエクソソームが細胞動態にあたえる影響を検証した。transwell migration assayを行ったところ、21%酸素濃度の場合と比べて、1%酸素濃度で培養したMSCsから回収されたエクソソームの方が、ラットMSCsおよびヒトMSCsの遊走細胞数が多かった。また、tube formation assayを行ったところ、21%酸素濃度の場合と比べて、1%酸素濃度で培養したMSCsから回収されたエクソソームの方が、HUVECsの管腔形成が亢進し、分岐部が増加した。
    ③幹細胞由来エクソソーム中に含まれる核酸へ低酸素がおよぼす影響を検証した。21%と1%のそれぞれの酸素濃度で培養したMSCs由来エクソソームは、小胞内に含有している核酸のプロファイリングが異なっていた。
    これらin vitro実験の研究成果をまとめて、国際学会(EAO Congress 2019 - 28th Annual Scientific Meeting:2019年9月葡国)で発表した。
    初年度に計画していた研究内容は計画どおりおおむね実施できた。初年度の実験結果により低酸素環境によって、エクソソームの分泌量が増大し、粒径も増大することがあきらかとなった。低酸素条件のエクソソームを投与されたラット骨髄由来間葉系幹細胞は増殖能が亢進し、ヒト臍帯静脈内皮細胞の管腔形成が亢進した。今後は、エクソソーム内の核酸のプロファイリング、クラスター解析をさらにすすめ、低酸素条件があたえる影響を評価する。ラット頭蓋骨骨欠損モデルを用いた動物実験で骨再生能を評価することで、低酸素条件が幹細胞の分泌物に与える影響を評価していく。
    ■複数の低酸素培養条件で回収したMSCs-Exo内の核酸の変化を検証する。MSCs-Exoを投与した培地で培養したMSCsからRNAを回収した後、マイクロアレイを行いmiRNA発現プロファイリングを行う。作成されたmiRNAプロファイルどうしを比較して、3倍以上高く発現していたものを骨再生に有効なmiRNA候補として選定する。
    ■複数の低酸素培養条件を設定してMSCs-Exoを回収し、ラット頭蓋骨骨欠損モデルへ投与して骨再生能を比較する。Wistar/ST ラット8 週齡雄にトレフィンバーを用いて頭蓋骨骨欠損を作成しMSCs-Exo をアテロコラーゲンスポンジ(テルダーミス)を足場として欠損作成部位に投与する。対照群にはPBS を足場に含浸させて投与する。投与後8 週にて検体を回収し評価する。μCT を撮影しX 線学的に、組織切片を作成して組織学的に骨形成量を評価する。川本法により凍結切片を作成し、骨再生機序に関して免疫組織化学的に評価する。CD44 やCD90、CD31 に対する抗体を用い内在性幹細胞の遊走や血管新生などを評価する。MSCs-Exo の投与部位からRNA を回収して、Realtime RT-PCR を行い骨形成関連遺伝子、血管新生関連遺伝子の発現を評価し骨再生機序を検証する。
    ■高い再生能を有するMSCs-Exo内で高発現であったmiRNAの機能解析を行う。上記で選定されたmiRNAに関してmiRNAの機能解析として、miRNAインヒビターを用いmiRNAの機能を抑制、他方でpCMV-MIRプラスミドを用いmiRNAを過剰発現させて、骨形成関連遺伝子発現や細胞動態の変化、ラット頭蓋骨欠損モデルで骨再生能を検証する。