| Project/Area Number |
02454438
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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 |
Conservative dentistry
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| Research Institution | Kyushu University |
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Principal Investigator |
NAGASAWA Hisashi Kyushu University, Professor, 歯学部, 教授 (10013848)
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| Co-Investigator(Kenkyū-buntansha) |
NAKASHIMA Misako Kyushu University., 歯学部, 助手 (20207773)
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| Project Period (FY) |
1990 – 1991
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| Project Status |
Completed (Fiscal Year 1991)
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| Budget Amount *help |
¥6,800,000 (Direct Cost: ¥6,800,000)
Fiscal Year 1991: ¥1,700,000 (Direct Cost: ¥1,700,000)
Fiscal Year 1990: ¥5,100,000 (Direct Cost: ¥5,100,000)
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| Keywords | pulp cell / odontoblast / growth factor / differentiation factor / phosphophoryn / osteocalcin / pulpotomy / Bone Morphogenetic Protein / 骨形成因子 / 歯髄培養細胞 / DNA合成 / アルカリフォスファタ-ゼ / 細胞増殖 / 細胞分化 / 蛍光抗体法 / III型コラ-ゲン / 歯髄 / 象牙質 / 初代培養 / 高速液体クロマトグラフィ- |
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| Research Abstract |
Crude bone morphogenetic protein (BMP) extracted from bovine demineralized dentine was gel filtrated by high performance liquid chromatography. The fractions which stimulated DNA synthesis, inhibited it and stimulated alkaline phosphatase activity on pulp cells in culture could be separated, suggesting that the growth factor and dentine inducible factor may be not identical. Insulin-like growth factor I, II which has known contained in dentine stimulated pulp cell growth. We, therefore, tried to isolate dentine inducible factor as follows. An effluent fraction by 0.15-0.50M NaCl in heparin sepharose affinity chromatography was implanted subcutaneously in rat. Large amount of bone with bone marrow which was high in alkaline phosphatase activity was induced. The effluent fraction increased dose-dependently immunofluorescent staining using phosphophoryn antibody on day 14 in pulp cell culture. Northern blot analysis revealed that the effluent fraction also increased the expression of osteocalcin mRNA compared with other fractions.
Heparin sepharose affinity chromatography might be effective to separate dentine inducible factor from crude BMP. We are now under way to develope more effective and convenient assay method.
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