| Project/Area Number |
62480403
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| Research Category |
Grant-in-Aid for General Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Research Field |
外科・放射線系歯学
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| Research Institution | Tokyo Medical and Dental University |
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Principal Investigator |
ENOMOTO Syoji The 2nd Dept. of Oral and Maxillofacial Surgery Faculty of Dentistry, Professor, 歯学部, 教授 (40013940)
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| Co-Investigator(Kenkyū-buntansha) |
ISHIDA Satoshi Dept. of Clinic for Initial Diagnosis Faculty of Dentistry, Instructor, 歯学部, 助手 (00134734)
NAGURA Hideaki The 2nd Dept. of Oral and Maxillofacial Surgery Faculty of Dentistry, Assistant, 歯学部, 助教授 (80013960)
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| Project Period (FY) |
1987 – 1988
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| Project Status |
Completed (Fiscal Year 1988)
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| Budget Amount *help |
¥5,600,000 (Direct Cost: ¥5,600,000)
Fiscal Year 1988: ¥1,500,000 (Direct Cost: ¥1,500,000)
Fiscal Year 1987: ¥4,100,000 (Direct Cost: ¥4,100,000)
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| Keywords | BMP / Atelocollagen / Hydroxyapatite / Diffusion chamber / Bone marrow / Agarose gel / 骨誘導 / ディフュージョンチャンバー / 骨形成蛋白 / 人工骨 / 医療用コラーゲン |
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| Research Abstract |
1) Purification of BMP: A crude BMP, which was extracted from decalcified bovine bone, was separated with gel filtration (Sephacryl S-200 HR) and cation exchange (S-Sepharose Fast Flow) chromatography. Then additional separation was performed with heparin affinity chromatography (Shodex SF-Pack HR-894). The electrophoretic examination of the final BMP-fraction revealed a smear band appearing between 20 and 30 K of molecular weight. Therefore the molecular weight of BMP has not yet been able to be determined. We are now purifying the pure BMP by additional use of reverse phase HPLC.
2) Examination of the materials mixed with BMP: Since the yield of BMP is very small, we looked for a carrier which enhances the BMP activity. As far as we examined, atelocollagen was the best material as a carrier of BMP and the most suitable for clinical application.
3) Examination of the preparation of BMP-complex: Atelocollagen-BMP or Hydroxyapatite-atelocollagen-BMP complcx was implanted subcutancously in
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to rats. When these BMP-complex were lyophillzed and compressed, new bone formation was induced the best and we could give our desired form to the implant materials.
4) Study of BMP using diffusion chambers: When bone marrow cells were implanted intraperitoneally within diffusion chambers with BMP, cartilage and bone formation was observed. In addition, when marrowderived fibroblast-like cells, which were acquired by the cultivation of marrow cells and were considered to be derived from marrow stroma, were inplanted as described above, cartilage and bone formation was detected. Therefore it was suspected that the target cells of BMP were derived not from hemopoietic cells, but from marrow stroma.
5) Effects of BMP in vitro: When mesenchymal cells derived from skeletal muscle were cultured in agarose gel and treated with partially purified BMP, induction of chondrocyte-like colony-formation was detected. The similar chondrocyte-like colony-forming activity was present in a dentin extract. However, whether BMP is the same factor that induces chondrocyte-like colony formation in agarose gel is the problem under discussion. Therefore we are now checking the differences between cartilage-inducing activity in agarose gel culture and BMP besides the purification of BMP. Less
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