Belarusian Medical Journal (in press)

APPLICATION OF 3-DIMENSIONAL MATRIX WITH CHONDROCYTES FOR TREATMENT OF ARTICULAR CARTILAGE DEFECTS
V.A. Goranov, Y.A. Haranava, A.A. Arabey, S.I. Tretyak, O.L. Eismont, and E.D. Belaenko

Chondrocytes isolated from tissue possess higher rates of proliferation, differentiation and cooperation when forming mature tissue than the cells obtained by puncture biopsy. The latter cells are rather stable during their growth in clustered culture.

Using of collagen matrix provides rapid chondrocytes growth. Specially stabilized polypeptide matrix used to confine chondrocytes is optimal to form chondroid structure as it provides uniform 3D-distribution of the cells and maintains proper organotypical structure. Moreover, chondrocytes turned to produce proper extracellular matrix.

The use of growth factors and other supplements helps to maintain culture stability. To avoid cell dedifferentiation and stimulate their proliferation we tested several substances like ascorbic acid, insulin, transferrin, FGF, TGF, and others. Combination of TGF, ascorbic acid and transferrin proved to be the best choice.

After first 6 days of cultivation in polypeptide matrix we observed small hyaline-like clusters, and by 10-14 days of cultivation chondrocytes differentiation was in fact complete, with hyaline-like clusters being of up to 2mm in size. Alcian blue and Safranin O staining demonstrated high level of proteoglycans production.