合成微小環境を用いた軟骨組織のｉｎ ｖｉｔｒｏ構築

• Project/Area Number: 14F04106
• Research Category: Grant-in-Aid for JSPS Fellows
• Allocation Type: Single-year Grants
• Section: 外国
• Research Field: Dental engineering/Regenerative dentistry
• Research Institution: Okayama University
• Principal Investigator: 松本 卓也 岡山大学, 医歯(薬)学総合研究科, 教授 (40324793)
• Co-Investigator(Kenkyū-buntansha): HARA EMILIO 岡山大学, 医歯(薬)学総合研究科, 外国人特別研究員 ; HARA Emilio 岡山大学, 医歯(薬)学総合研究科, 外国人特別研究員
• Project Period (FY): 2014-04-25 – 2016-03-31
• Project Status: Completed (Fiscal Year 2015)
• Budget Amount: ¥2,400,000 (Direct Cost: ¥2,400,000); Fiscal Year 2015: ¥1,200,000 (Direct Cost: ¥1,200,000); Fiscal Year 2014: ¥1,200,000 (Direct Cost: ¥1,200,000)
• Keywords: 軟骨 / 合成微小環境 / DNAメチル化 / 石灰化

Outline of Annual Research Achievements

The purpose of this study was to identify new methods and culture conditions to enhance chondrogenic differentiation of human bone marrow-derived mesenchymal stem/progenitor cells (hBMSCs) by applying chemical, physical and biological cues, including overexpression of DNA methyltransferases (DNMTs) in cells. We attempted to fabricate a chemically- and physically-controlled microenvironment using materials (e.g., hydrogels) that recapitulate native cell niche characteristics to optimally modulate the chondrogenic differentiation of BMSCs. Besides the physical and chemical stimulations, we also evaluated the chondrogenic differentiation of hBMSCs after overexpression of DNMTs that eventually induce DNA methylation at CPG rich regions of key genes. To study the effect of DNMTs on chondrogenesis of hBMSCs, we transfected overexpression vectors of DNMT3A, DNMT3B and the pcDNA control in hBMSCs, and culture the cells in a 3D micromass culture system with growth factors and glucocorticoids.
Additionally, since in vitro synthesized cartilage tissue generally undergoes through mineralization, we also attempted to understand the mechanisms involved in the chondrocyte death associated with initial mineralization process by using the secondary ossification center of mouse femur as in vivo model.

Research Progress Status

27年度が最終年度であるため、記入しない。

Strategy for Future Research Activity

27年度が最終年度であるため、記入しない。

Research Products

• [Int'l Joint Research] Harvard Univerisity(米国)
• [Journal Article] Antagonistic Effects of Insulin and TGF-β3 during Chondrogenic Differentiation of Human BMSCs under a Minimal Amount of Factors. (2016)
  • Author(s): Emilio Satoshi Hara, Ono Mistuaki, Yoshioka Yuya, Ueda Junji, Hazehara Yuri, Hai Thanh Pham, Matsumoto Takuya, Kuboki Takuo.
  • Journal Title: Cells Tissues Organs Volume: 201 Issue: 2 Pages: 88-96
  • DOI: 10.1159/000442411
  • Peer Reviewed / Open Access / Int'l Joint Research / Acknowledgement Compliant
• [Journal Article] Fluocinolone acetonide is a potent synergistic factor of TGF-β3-associated chondrogenesis of bone marrow-derived mesenchymal stem cells for articular surface regeneration (2015)
  • Author(s): Emilio Satoshi Hara, Mitsuaki Ono, Pham Thanh Hai, Wataru Sonoyama, Satoshi Kubota, Masaharu Takigawa, Takuya Matsumoto, Marian F. Young, Bjorn R. Olsen, Takuo Kuboki
  • Journal Title: J. Bone Miner. Res. Volume: 未定 Issue: 9 Pages: 1585-1596
  • DOI: 10.1002/jbmr.2502
  • Peer Reviewed / Open Access
• [Journal Article] Mesenchymal stem/progenitor cell isolation from tooth extraction sockets (2014)
  • Author(s): Ryu Nakajima, Mitsuaki Ono, Emilio Satoshi Hara, Yasutaka Oida, Shigehiko Shinkawa, Hai Thanh Pham, Kentaro Akiyama, Wataru Sonoyama, Kenji Maekawa, Takuo Kuboki
  • Journal Title: Journal of Dental Research Volume: 93 Issue: 11 Pages: 1133-1140
  • DOI: 10.1177/0022034514549377
  • Peer Reviewed
• [Journal Article] Comparison of platelet serotonin transporter activity in subjects with severe sleep bruxism and control (2014)
  • Author(s): Hajime Minakuchi, Chiharu Sogawa C, Emilio Satoshi Hara, Haruna Miki, Kenji Maekawa, Norio Sogawa, Shigeo Kitayama, Yoshizo Matsuka, Glenn T Clark, Takuo Kuboki
  • Journal Title: Journal of Prosthodontic Research Volume: 58 Issue: 4 Pages: 217-222
  • DOI: 10.1016/j.jpor.2014.06.003
  • NAID: 130004714111
  • ISSN: 1883-1958, 1883-9207
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] A short-term treatment with tumor necrosis factor-alpha enhances stem cell phenotype of human dental pulp cells (2014)
  • Author(s): Mayu Ueda, Takuo Fujisawa, Mitsuaki Ono, Emilio Satoshi Hara, Hai Thanh Pham, Ryu Nakajima, Wataru Sonoyama, Takuo Kuboki
  • Journal Title: Stem Cell Research & Therapy Volume: 5 Issue: 1 Pages: 31-31
  • DOI: 10.1186/scrt420
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] The regenerative effects of CCN2 independent modules on chondrocytes in vitro and osteoarthritis models in vitro (2014)
  • Author(s): Tarek Abd El Kader, et al.
  • Journal Title: Bone Volume: 59 Pages: 180-188
  • DOI: 10.1016/j.bone.2013.11.010
  • Peer Reviewed / Open Access
• [Journal Article] Efficient bone formation in a swine socket lift model using Escherichia coli-derived recombinant human bone morphogenetic protein-2 adsorbed in β-tricalcium phosphate. (2014)
  • Author(s): Mitsuaki Ono, Wataru Sonoyama, Katushi Yamamoto, Yasutaka Oida, Kentaro Akiyama, Shigehiko Shinkawa, Ryu Nakajima, Hai Than Pham, Emilio Satoshi Hara, Takuo Kuboki
  • Journal Title: Cells, Tissues, Organs Volume: 199 Issue: 4 Pages: 249-255
  • DOI: 10.1159/000369061
  • Peer Reviewed / Open Access
• [Presentation] Fluocinolone acetonide is a potent synergistic factor of TGF-β3-associated chondrogenesis of bone marrow-derived mesenchymal stem cells for articular surface regeneration. (2016)
  • Author(s): Hara ES, Ono M, Pham HT, Sonoyama W, Kubota S, Takigawa M, Matsumoto T, Young MF, Olsen BR, Kuboki T.
  • Organizer: 第29回日本軟骨代謝学会
  • Place of Presentation: 広島
  • Year and Date: 2016-02-20
• [Presentation] Potent cartilage regeneration with fluocinolone acetonide and TGF-β3. (2015)
  • Author(s): Hara ES, Ono M, Pham HT, Kubota S, Takigawa M, Young MF, Olsen BR, Matsumoto T, Kuboki T.
  • Organizer: 第37回日本バイオマテリアル学会大会
  • Place of Presentation: 京都
  • Year and Date: 2015-11-09
• [Presentation] A high-throughput screening identified fluocinolone acetonide as a potent synergistic factor of TGF-β3-mediated chondrogenesis of BMSCs for articular surface repair. (2015)
  • Author(s): Hara ES, Ono M, Pham HT, Sonoyama W, Kubota S, Takigawa M, Young MF, Olsen BR, Matsumoto T, Kuboki T.
  • Organizer: American Society for Bone and Mineral Research Annual Meeting
  • Place of Presentation: シアトル, USA
  • Year and Date: 2015-10-12
  • Int'l Joint Research
• [Presentation] A Novel Method to Induce Strong Chondrogenesis of BMSCs and Promote Articular Surface Repair using Fluocinolone Acetonide and TGF-β3. (2015)
  • Author(s): Hara ES, Ono M, Pham HT, Sonoyama W, Kubota S, Takigawa M, Young MF, Olsen BR, Matsumoto T, Kuboki T.
  • Organizer: Biomedical Engineering Society Annual Meeting
  • Place of Presentation: タンパ, USA
  • Year and Date: 2015-10-08
  • Int'l Joint Research
• [Presentation] Qualitative analysis of tissue mineralization during secondary ossification of mouse femur (2014)
  • Author(s): Emilio Satoshi Hara, Masahiro Okada, Takako Hattori, Satoshi Kubota, Takuo Kuboki, TakuyaMatsumoto
  • Organizer: 第36回日本バイオマテリアル学会大会
  • Place of Presentation: 東京
  • Year and Date: 2014-11-18 – 2014-11-19
• [Presentation] Novel insights into the mineralization process during secondary ossification of the cartilage (2014)
  • Author(s): Emilio Satoshi Hara, Masahiro Okada, Takako Hattori, Satoshi Kubota, Takuo Kuboki, Takuya Matsumoto
  • Organizer: International Symposium 2014 - Foresight Program "Nano-Biomaterials and Regenerative Medicine"
  • Place of Presentation: 東京
  • Year and Date: 2014-10-08 – 2014-10-09
• [Presentation] Quantitative analysis of mineralized tissue formation during cartilage secondary ossification of mouse femur (2014)
  • Author(s): Emilio Satoshi Hara, Masahiro Okada, Takuya Matsumoto
  • Organizer: 第64回日本歯科理工学会学術講演会
  • Place of Presentation: 広島
  • Year and Date: 2014-10-05 – 2014-10-06