Development of tissue regeneration method applying mechanotransduction and stem cell migration mechanism

• Project/Area Number: 18K09635
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Review Section: Basic Section 57040:Regenerative dentistry and dental engineering-related
• Research Institution: Nagoya University
• Principal Investigator: FUJIO Masahito 名古屋大学, 医学系研究科, 助教 (90612804)
• Co-Investigator(Kenkyū-buntansha): 土屋 周平 名古屋大学, 医学部附属病院, 助教 (20569785)
• Project Period (FY): 2018-04-01 – 2021-03-31
• Project Status: Completed (Fiscal Year 2020)
• Budget Amount: ¥3,900,000 (Direct Cost: ¥3,000,000、Indirect Cost: ¥900,000); Fiscal Year 2020: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000); Fiscal Year 2019: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000); Fiscal Year 2018: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000)
• Keywords: 骨再生 / 幹細胞 / メカノトランスダクション / 骨延長

Outline of Final Research Achievements

Human bone marrow-derived mesenchymal stem cells and human periodontal ligament-derived fibroblasts were cultured by applying cyclic stretch stimulation. The conditioned media was collected and the secretome were analyzed. Angiogenesis-related factors (VEGF, PDGF-AA) and osteogenic factors (BMP-2, BMP-4) were released more than when cultured without applying cyclic stretch stimulation. An increase in angiogeneis and osteogenesis was observed in vitro and in vivo. It was suggested that by changing the cell culture environment, a disease-specific conditioned medium could be obtained.

Academic Significance and Societal Importance of the Research Achievements

失われた骨組織の再生方法として、細胞移植の研究が盛んに行われている。我々はこれまでヒト間葉系幹細胞（hMSC）を骨欠損部に移植し良好な結果を得てきたが，細胞採取時の侵襲や感染のリスク、細胞の腫瘍化などの問題点も指摘されている。細胞移植に代わる方法として幹細胞培養後の培養上清（conditioned medium以下CM）を用いる方法を考案した。本研究では、細胞に機械的刺激を加えることにより、CMをより骨再生に適したものにできる可能性を示した。

Research Products

• [Journal Article] Conditioned media from mesenchymal stromal cells and periodontal ligament fibroblasts under cyclic stretch stimulation promote bone healing in mouse calvarial defects (2020)
  • Author(s): Ogisu Kota、Fujio Masahito、Tsuchiya Shuhei、Tsuboi Makoto、Qi Chang、Toyama Naoto、Kamio Hisanobu、Hibi Hideharu
  • Journal Title: Cytotherapy Volume: 22 Issue: 10 Pages: 543-551
  • DOI: 10.1016/j.jcyt.2020.05.008
  • Peer Reviewed
• [Presentation] 伸展刺激下で得た培養細胞上清における骨形成・血管新生能の検討 (2019)
  • Author(s): 荻須宏太、藤尾正人、土屋周平、岡部一登、外山直人、日比英晴
  • Organizer: 第６４回公益社団法人 日本口腔外科学会総会・学術大会
• [Presentation] 伸展刺激下で得た細胞培養上清における骨形成・血管新生能の検討 (2019)
  • Author(s): 荻須宏太・藤尾正人・土屋周平・岡部一登・神尾尚伸・外山直人・日比英晴
  • Organizer: 日本口腔科学会