| Project/Area Number |
61440082
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| Research Category |
Grant-in-Aid for General Scientific Research (A)
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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 | Hokkaido University |
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Principal Investigator |
ASAAKI Kawamura Hokkaido University, School of Dentistry, 歯学部, 教授 (80001014)
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| Co-Investigator(Kenkyū-buntansha) |
ADASHI Mikoya Hokkaido University, School of Dentistry, 歯学部, 助手 (10181869)
OHSIKATSU Fujiwara Hokkaido University, School of Dentistry, 歯学部, 助手 (50142740)
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| Project Period (FY) |
1986 – 1987
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| Project Status |
Completed (Fiscal Year 1987)
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| Budget Amount *help |
¥20,800,000 (Direct Cost: ¥20,800,000)
Fiscal Year 1987: ¥4,800,000 (Direct Cost: ¥4,800,000)
Fiscal Year 1986: ¥16,000,000 (Direct Cost: ¥16,000,000)
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| Keywords | Calcification / Protease of Matrix Vesicles / Culture System / Calcifying Matrices / コラーゲン / 石灰化 / 基質小胞 / タンパク分解酵素 / プロテオグリカン |
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| Research Abstract |
In this study, in order to analyse the proteases of matrix vesicles in calcification, we aimed to establish the culture system in which the process just like in vivo calcification was expressed.
For its purpose, we used the established osteogenic cells, MC3T3-El which possessed high collagen synthesis activity, matrix vesicle-producing activity and calcification activity.
Primarily we examined definite culture condition and compatibility of FCS to cells. Secondarily we confirmed that calcified tissues produced by El cells were composed of hydroxyapatite by electron diffraction analysis and energy disperse x-ray analysis. Finally we analysed biochemical changes in this culture system. Consequently we observed the increase of AL-P activity, the decrease of hexosamines and the synthesis of type 1 collagen. It was markedly important that crosslining of collagen in this culture was indicated as same as in vivo and El cells produced matrix vesicle-like substances. It was suggested to be of capability to analyse proteases of matrix vesicle enzymologically in this culture system.
Now we are planning to establish the culture system of chondrocytes in epiphyseal cartilage and investigate in detail the qualitative changes of collagen, interaction of noncollagenous components including proteoglycan with it, and significance of proteases of matrix versicles in calcification.
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