Development of functional multilayered mesenchymal stem cells sheet for lumbar fusion surgery

• Project/Area Number: 17K10982
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Orthopaedic surgery
• Research Institution: Kitasato University
• Principal Investigator: Inoue Gen 北里大学, 医学部, 准教授 (80594209)
• Co-Investigator(Kenkyū-buntansha): 内田 健太郎 北里大学, 医学部, 講師 (50547578) ; 馬渕 洋 東京医科歯科大学, 大学院医歯学総合研究科, 助教 (50424172) ; 宮城 正行 北里大学, 医学部, 講師 (90627556) ; 松下 治 岡山大学, 医歯薬学総合研究科, 教授 (00209537)
• Project Period (FY): 2017-04-01 – 2020-03-31
• Project Status: Completed (Fiscal Year 2019)
• Budget Amount: ¥4,680,000 (Direct Cost: ¥3,600,000、Indirect Cost: ¥1,080,000); Fiscal Year 2019: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000); Fiscal Year 2018: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000); Fiscal Year 2017: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000)
• Keywords: 細胞シート / 積層 / 脊椎固定術 / 骨癒合 / 骨形成タンパク / 後側方固定術 / 移植・再生医療 / 細胞・組織

Outline of Final Research Achievements

Cell-based regenerative therapy has the potential to promote bone formation in spinal fusion. Here, we developed a multilayered cell-based bone formation system using cells coated with fibronectin-gelatin (FN-G) nanofilms. The multilayered human mesenchymal stem cells (hMSCs) were formed after two days of culture and were shown to express higher levels of BMP-2, TGF-β, bFGF and VEGF compared to monolayer cultures of hMSCs. The MLMCs were used as a graft material in combination with a collagen-binding bFGF and BMP-2. In grafted with the hMSCs, significantly higher levels of callus volume and bone mineral content were observed compared to the sham controls. The callus volume and bone mineral content were further increased in situ grafted with bFGF-PKD-CBD/BMP-2/hMSCs. Taken together, these results suggest that bFGF-PKD-CBD/BMP-2/hMSC may offer novel therapeutic approaches to augment spinal fusion.

Academic Significance and Societal Importance of the Research Achievements

高齢社会を迎えた我が国において、高齢者の脊椎固定術が確実に増加している。また、高齢者は若年者に比べ骨癒合が遷延することが知られている。治療の長期化に伴う患者の社会復帰の遅延や、続発する合併症は、患者に肉体的・精神的苦痛を与えるとともに医療費の増加と社会経済的損失に直結する。従って、本シーズは国民の健康寿命の延長に向けて重要な意義を持つ。

Research Products

• [Journal Article] Effect of single injection of recombinant human bone morphogenetic protein-2-loaded artificial collagen-like peptide in a mouse segmental bone transport model (2019)
  • Author(s): Tazawa R, Minehara H, Matsuura T, Kawamura T, Uchida K, Saito W, Inoue G, Takaso M.
  • Journal Title: Biomed Res Int Volume: - Pages: 1014594-1014594
  • DOI: 10.1155/2019/1014594
  • Peer Reviewed / Open Access
• [Journal Article] Wrapping with Basic Fibroblast Growth Factor-Impregnated Collagen Sheet Reduces Rat Sciatic Nerve Allodynia (2019)
  • Author(s): Mukai M, Uchida K, Hirosawa N, Murakami K, Kuniyoshi K, Inoue G, Miyagi M, Sekiguchi H, Shiga Y, Inage K, Orita S, Suzuki T, Matsuura Y, Takaso M, Ohtori S
  • Journal Title: J Orthop Res Volume: 37 Issue: 10 Pages: 2258-2263
  • DOI: 10.1002/jor.24349
  • Peer Reviewed
• [Journal Article] The impact of switching once-weekly teriparatide to denosumab in osteoporosis patients (2019)
  • Author(s): Miyagi M, Fujimaki H, Naruse K, Suto K, Inoue G, Nakazawa T, Imura T, Saito W, Uchida K, Shirasawa E, Takahira N, Takaso M
  • Journal Title: J Orthop Sci Volume: 24 Issue: 1 Pages: 153-158
  • DOI: 10.1016/j.jos.2018.08.001
  • Peer Reviewed / Open Access
• [Journal Article] Nerve growth factor continuously elevates in a rat rotator cuff tear model. (2019)
  • Author(s): Nagura N, Kenmoku T, Uchida K, Nakawaki M, Inoue G, Takaso M
  • Journal Title: J Shoulder Elbow Surg Volume: 28 Issue: 1 Pages: 143-148
  • DOI: 10.1016/j.jse.2018.06.030
  • Peer Reviewed / Open Access
• [Journal Article] Total en bloc spondylectomy of the eleventh thoracic vertebra following denosumab therapy for the treatment of a giant cell tumor. (2017)
  • Author(s): Inoue G, Imura T, Miyagi M, Saito W, Tazawa R, Nakazawa T, Takaso M.
  • Journal Title: Oncol Lett Volume: 14 Issue: 4 Pages: 4005-4010
  • DOI: 10.3892/ol.2017.6655
  • Peer Reviewed
• [Journal Article] Basic Fibroblast Growth Factor-Anchored Multilayered Mesenchymal Cell Sheets Accelerate Periosteal Bone Formation (2017)
  • Author(s): Uchida K, Inoue G, Matsushita O, Horikawa K, Sekiguchi H, Saito W, Takano S, Fujimaki H, Miyagi M, Takaso M.
  • Journal Title: Biomed Res Int Volume: 4371460 Pages: 1-8
  • DOI: 10.1155/2017/4371460
  • Peer Reviewed / Open Access
• [Journal Article] Effect of Freeze-Dried Allograft Bone with Human Basic Fibroblast Growth Factor Containing a Collagen-Binding Domain From Clostridium Histolyticum Collagenase on Bone Formation After Lumbar Posterolateral Fusion Surgery in Rats (2017)
  • Author(s): Inoue G, Uchida K, Matsushita O, Fujimaki H, Saito W, Miyagi M, Sekiguchi H, Nishi N, Ohtori S, Yogoro M, Takaso M.
  • Journal Title: Spine Volume: 印刷中 Issue: 17 Pages: 995-1001
  • DOI: 10.1097/brs.0000000000002074
  • Peer Reviewed / Acknowledgement Compliant
• [Presentation] コラーゲン材料とコラーゲン結合型成長因子を用いた運動器再生 (2019)
  • Author(s): 内田健太郎、松下 治、井上 玄、美間健彦、高相晶士
  • Organizer: 第51回 日本結合組織学会学術大会
• [Presentation] ラット坐骨神経損傷モデルに対する塩基性繊維芽細胞増殖因子添加コラーゲンシートの痛覚過敏抑制効果とHO-1誘導能の検討 (2019)
  • Author(s): 向井務晃、内田健太郎、廣澤直也、村上賢一、稲毛一秀、折田純久、松浦祐介、井上玄、國吉一樹、高相晶士、大鳥精司．
  • Organizer: 第34回日本整形外科学会基礎学術集会
• [Presentation] 骨形成蛋白（BMP-2）含有人工コラーゲンゲルはマウス難治性骨折モデルにおける骨形成と骨癒合を促進する (2019)
  • Author(s): 関口裕之、内田健太郎、井上玄、庄司真太郎、宮城正行、齋藤亘、大貫裕子、高相晶士
  • Organizer: 第34回日本整形外科学会基礎学術集会
• [Presentation] 塩基性線維芽細胞増殖因子と局所硬化デキストランゲルを用いた新規骨形成促進法の有用性 (2019)
  • Author(s): 庄司真太郎、内田健太郎、関口裕之、井上 玄、齋藤 亘、宮城正行、村田幸佑、川久保 歩、高相晶士
  • Organizer: 第33回日本整形外科学会基礎学術集会
• [Presentation] 難治性骨折に対する新規コラーゲン結合型塩基性線維芽細胞増殖因子/人工コラーゲン複合体の有用性の検討 (2017)
  • Author(s): 関口裕之、内田健太郎、井上玄、宮城正行、相川淳、高野昇太郎、名倉直重、中脇充章、庄司真太郎、大貫裕子、高相昌士
  • Organizer: 第32回日本整形外科学会基礎学術集会
• [Presentation] コラーゲン結合型塩基性線維芽細胞増殖因子を用いた末梢神経再生の試み (2017)
  • Author(s): 藤巻寿子、井上玄、内田健太郎、根本典子、宮城正行、大鳥精司、高相昌士
  • Organizer: 第32回日本整形外科学会基礎学術集会