Development of osteoblast culture system for bone regenerative treatment for cell transplantation

• Project/Area Number: 16K20684
• Research Category: Grant-in-Aid for Young Scientists (B)
• Allocation Type: Multi-year Fund
• Research Field: Periodontology
• Research Institution: Aichi Gakuin University
• Principal Investigator: Aino Makoto 愛知学院大学, 歯学部, 講師 (20572811)
• Project Period (FY): 2016-04-01 – 2019-03-31
• Project Status: Completed (Fiscal Year 2018)
• Budget Amount: ¥2,600,000 (Direct Cost: ¥2,000,000、Indirect Cost: ¥600,000); Fiscal Year 2017: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000); Fiscal Year 2016: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000)
• Keywords: 歯周病 / 骨再生 / 骨芽細胞 / ３次元培養 / 細胞外環境 / 再生医療

Outline of Final Research Achievements

In recent years, three-dimensional culture with a hydrogel that is flexible and easily mimics a living environment for cell transplantation treatment has been actively studied in recent years. The research representative examined the influence of the culture environment on the cells by performing three-dimensional culture using Gelatin Metacryloyl (GelMA), which is a highly compatible and highly cell-adhesive biomaterial. Since GelMA changes hardness by concentration, we analyzed hardness optimized for osteoblasts, and as a result, 16 Kpa was optimum. As a result of culturing at this hardness, the cells aggregated and an increase in expression of bone matrix protein gene was observed. From these facts, it was found that three-dimensional culture by GelMA is suitable for culture toward bone tissue regeneration.

Academic Significance and Societal Importance of the Research Achievements

歯周病は日本国民が最も歯を失う原因となる疾患である。この歯周病で破壊された骨組織は一般的な歯科治療で回復できる症例が限られており、また患者自身が自覚症状を持つほどに進行した歯周病では回復させる事が非常に難しい。現在多くの分野の医療において回復させる事が難しいような損傷を負った場合、細胞移植治療による回復が行われつつあるが、組織によって培養法や条件が異なる。今回の研究成果から骨組織の再生に適した細胞培養の可能性を示し、将来的には細胞移植治療につなげる事が可能になると考えられる。

Research Products

• [Journal Article] IL-15 and RANKL Play a Synergistically Important Role in Osteoclastogenesis. (2017)
  • Author(s): Okabe I, Kikuchi T, Mogi M, Takeda H, Aino M, Kamiya Y, Fujimura T, Goto H, Okada K, Hasegawa Y, Noguchi T, Mitani A.
  • Journal Title: J Cell Biochem Volume: 118(4) Issue: 4 Pages: 739-747
  • DOI: 10.1002/jcb.25726
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] F-spondin negatively regulates dental follicle differentiation through the inhibition of TGF-beta activity. (2017)
  • Author(s): Orimoto A, Kurokawa M, Handa K, Ishikawa M, Nishida E, Aino M, Mitani A, Ogawa M, Tsuji T Saito M
  • Journal Title: Arch Oral Biol Volume: 79 Pages: 7-13
  • DOI: 10.1016/j.archoralbio.2017.02.019
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Effect of interleukin (IL)-35 on IL-17 expression and production by human CD4+ T cells. (2016)
  • Author(s): Okada K, Fujimura T, Kikuchi T, Aino M, Kamiya Y, Izawa A, Iwamura Y, Goto H, Okabe I, Miyake E, Hasegawa Y, Mogi M, Mitani A.
  • Journal Title: Peer J. Volume: e2999 Pages: e2999-e2999
  • DOI: 10.7717/peerj.2999
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Serum Amyloid A Promotes E-Selectin Expression via Toll-Like Receptor 2 in Human Aortic Endothelial Cells (2016)
  • Author(s): Nishida E, Aino M, Kobayashi SI, Okada K, Ohno T, Kikuchi T, Hayashi JI, Yamamoto G, Hasegawa Y, Mitani A.
  • Journal Title: Mediators of Inflammation Volume: 7150509 Pages: 7150509-7150509
  • DOI: 10.1155/2016/7150509
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Presentation] 再生治療におけるGelMA-RFの足場材料としての新たな可能性について (2018)
  • Author(s): 後藤亮真,西田英作,小林周一郎,相野 誠,黒須康成,三谷章雄
  • Organizer: 第149回日本歯科保存学会秋季学術大会
• [Presentation] GelMA-RFの足場材料としての可能性について (2018)
  • Author(s): 後藤亮真,西田英作,小林周一郎,相野 誠,三谷章雄
  • Organizer: 第16回日本再生歯科医学会学術大会
• [Presentation] 歯周病病態におけるTh17細胞に対するIL-35の役割についての基礎的検討. (2017)
  • Author(s): 岡田康佑, 藤村岳樹, 菊池毅, 相野誠, 神谷洋介, 伊澤有郎, 後藤久嗣, 岩村侑樹, 岡部猪一郎, 三宅永莉子, 長谷川義明, 茂木眞希雄, 三谷章雄
  • Organizer: 第60回春季日本歯周病学会学術大会
• [Presentation] アルゴンイオンボンバードメントを用いたポリ塩化ビニリデン(PVDC)フィルムの生体活性増強とGBRへの応用. (2017)
  • Author(s): 小林周一郎, 林達秀, 相野誠, 西田英作, 朝倉正紀, 岡部栄治郎, 河合逹志, 野口俊英, 三谷章雄
  • Organizer: 第17回日本外傷歯学会学術大会
• [Presentation] ヒト歯肉上皮細胞におけるIL-35の影響について (2016)
  • Author(s): 岡田康佑, 藤村岳樹, 菊池毅, 相野誠, 神谷洋介, 伊澤有郎, 後藤久嗣, 岡部猪一郎, 三宅永莉子, 三谷章雄
  • Organizer: 第145回日本歯科保存学会秋季学術大会
  • Place of Presentation: 長野県松本市
  • Year and Date: 2016-10-27
• [Presentation] ヒト口腔由来上皮細胞のジルコニアに対する付着特性 (2016)
  • Author(s): 岡部栄治郎, 菊池毅, 石原裕一, 後藤久嗣, 相野誠, 河合逹志, 三谷章雄, 村上弘
  • Organizer: 第46回秋季日本口腔インプラント学会
  • Place of Presentation: 愛知県名古屋市
• [Presentation] Fundamental assessment of irradiation method for antimicrobial photodynamic therapy (aPDT) using a laser transmitted through soft tissue. (2016)
  • Author(s): Iwamura Y, Hayashi J, Sasaki Y, Okada K, Fujimura T, Nishida E, Aino M, Kikuchi T, Yamamoto H, Mitani A, Fukuda M.
  • Organizer: 15ht Congress of the World Federation for Laser Dentistry
  • Place of Presentation: 愛知県名古屋市
  • Int'l Joint Research
• [Presentation] Antimicrobial photodynamic therapy using a laser transmitted through soft tissue. (2016)
  • Author(s): Sasaki Y, Hayashi J, Iwamura Y, Okada K, Fujimura T, Nishida E, Aino M, Kikuchi T, Yamamoto H, Mitani A, Fukuda M.
  • Organizer: 15ht Congress of the World Federation for Laser Dentistry
  • Place of Presentation: 名古屋
  • Int'l Joint Research