Preclinical study of regenerative therapy for bone defects using dental pulp stem cells

• Project/Area Number: 20K10017
• 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: Tokyo Women's Medical University
• Principal Investigator: Kawase-Koga Yoko 東京女子医科大学, 医学部, 教授 (10392408)
• Co-Investigator(Kenkyū-buntansha): 飛田 護邦 順天堂大学, 革新的医療技術開発研究センター, 先任准教授 (10599038) ; 近津 大地 東京医科大学, 医学部, 主任教授 (30343122)
• Project Period (FY): 2020-04-01 – 2023-03-31
• Project Status: Completed (Fiscal Year 2022)
• Budget Amount: ¥4,420,000 (Direct Cost: ¥3,400,000、Indirect Cost: ¥1,020,000); Fiscal Year 2022: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000); Fiscal Year 2021: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000); Fiscal Year 2020: ¥1,040,000 (Direct Cost: ¥800,000、Indirect Cost: ¥240,000)
• Keywords: 歯髄幹細胞 / ミニブタ / 三次元顎骨再生 / GFP / 三次元骨再生法 / 顎骨再生 / 前臨床試験

Outline of Research at the Start

応募者らは、ヒト歯髄幹細胞と生理活性物質を組み合わせることによりこれまでにない短期間かつ高効率な骨芽細胞誘導法を開発し、マウス頭頂骨臨海骨欠損モデルにてその有効性を報告した。しかし、移植細胞の生体内での作用機序および大動物におけるヒト歯髄幹細胞由来骨芽細胞の骨分化誘導能は不明である。本研究では、GFPミニブタ歯髄幹細胞由来骨芽細胞を免疫不全ミニブタに移植し、生体内での作用を明らかにする。また、前臨床試験としてミニブタにおける広範囲顎骨モデルを作製し、応募者らが開発した高効率なヒト歯髄由来骨芽細胞法を用いてその有効性の評価を行い、顎骨再生手法を確立することを目指す。

Outline of Final Research Achievements

We have developed an efficient osteoblast induction method using human dental pulp stem cells in combination with biological active substances, and reported its efficacy in a mouse model of parietal bone defect. However, the in vivo mechanism of action of transplanted cells and the ability of pulp stem cell-derived osteoblasts to induce bone differentiation in large animals were unknown. In this study, we transplanted GFP minipig dental pulp stem cell-derived osteoblasts into a GFP(-) minipig jaw bone defect model and performed large animal experiments to clarify their mode of action in vivo. We succeeded in isolating and culturing the dental pulp stem cells from the minipigs and culturing them in three-dimensional culture, and confirmed significant osteogenic potential in the TH-added group.

Academic Significance and Societal Importance of the Research Achievements

ヒト歯髄幹細胞は、2000年にGronthosらがヒト歯髄に分化能の高い間葉系幹細胞が存在することを報告し（Proc Natl Acad Sci U S A, 2000）、新たな再生医療の細胞源として着目されている。前臨床試験として大動物の顎骨欠損モデルにおけるヒト歯髄幹細胞の有効性の検討という課題が残されており本研究を遂行した。本研究の成果は、われわれがこれまで行ってきたヒト歯髄幹細胞の高効率な骨分化誘導法の確立をさらに発展させるのみならず、再生工学、口腔外科学等、広い領域に波及効果をもたらすものであると確信している。そして後の臨床試験に必ずや繋がる研究である。

Research Products

• [Journal Article] Evaluation of Facial Soft Tissue in Patients With Cleft Lip and Palate Corrected Using Maxillary Protraction Appliances (2021)
  • Author(s): Uehara S, Honda A, Kawase-Koga Y, Chikazu D.
  • Journal Title: J Craniofac Surg. Volume: in press Issue: 4 Pages: 1480-1482
  • DOI: 10.1097/scs.0000000000007406
  • Peer Reviewed
• [Journal Article] Cell therapy for medication-related osteonecrosis of the jaw: update on treatment strategies (2021)
  • Author(s): N Kaibuchi, T Iwata, T Okamoto, Y Kawase-Koga, M Yamato
  • Journal Title: European Cells & Materials Volume: 41 Pages: 31-39
  • DOI: 10.22203/ecm.v041a03
  • Peer Reviewed / Open Access
• [Journal Article] Effects of Helioxanthin Derivative-Treated Human Dental Pulp Stem Cells on Fracture Healing (2020)
  • Author(s): D. Yamakawa, Y. Kawase-Koga, Y. Fujii, Y. Kanno, M. Sato, S. Ohba, Y. Kitaura, M. Kashiwagi, D. Chikazu
  • Journal Title: International Journal of Molecular Sciences Volume: 21 Issue: 23 Pages: 9158-9171
  • DOI: 10.3390/ijms21239158
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Bone Regeneration Potential of Human Dental Pulp Stem Cells Derived from Elderly Patients and Osteo-Induced by a Helioxanthin Derivative (2020)
  • Author(s): M. Sato, Y. Kawase-Koga, D. Yamakawa, Y. Fujii, D. Chikazu
  • Journal Title: International Journal of Molecular Sciences Volume: 21 Issue: 20 Pages: 7731-7745
  • DOI: 10.3390/ijms21207731
  • Peer Reviewed / Open Access
• [Journal Article] Less invasive approach for bone reconstruction using three-dimensional formable titanium mesh after removal of osseous lesions in the mandibular angle (2020)
  • Author(s): Kono M, Watanabe M, Hasegawa O, Kawase-Koga Y, Chikazu D.
  • Journal Title: Br J Oral Maxillofac Surg. Volume: 58 Issue: 9 Pages: e127-e129
  • DOI: 10.1016/j.bjoms.2020.06.028
  • Peer Reviewed
• [Journal Article] The Usefulness of Saturated Salt Solution Embalming Method for Oral Surgical Skills Training: A New Cadaveric Training Model for Bone Harvesting (2020)
  • Author(s): Watanabe M, Yoneyama Y, Hamada H, Kohno M, Hasegawa O, Takahashi H, Kawase-Koga Y, Matsuo A, Chikazu D, Kawata S, Itoh M.
  • Journal Title: Anat Sci Educ. Volume: 13 Issue: 5 Pages: 628-635
  • DOI: 10.1002/ase.1925
  • Peer Reviewed