Studies on the Mechanism of Calcification - On a Protease Associated with Matrix Vesicles -

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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.