Possible involvement of RGD (Arg-Gly-Asp)-containing extracellular matrix proteins in rat growth plate chondrocyte differentiation in culture.

Abstract

RGD (arg-gly-asp)-containing proteins have been shown to be components of cartilage matrix. In the present study, the role of interactions of cells with RGD-containing cartilage matrix proteins in rat costal epiphyseal chondrocyte differentiation was examined using a pelleted culture system as an in vitro model of endochondral ossification. Cell attachment assays showed the presence of integrins which mediated the binding of chondrocytes to fibronectin, a member of RGD-containing cartilage matrix proteins, in an RGD-dependent manner. In the early culture period, when chondrocytes had nonhypertrophic morphology with low levels of alkaline phosphatase, the exogenous addition of synthetic peptide GRGDSP (gly-arg-gly-asp-ser-pro) caused an increase in alkaline phosphatase levels and enlargement of chondrocytes in pelleted cultures. Treatment with GRGDSP from the early to late culture periods in association with the transition of chondrocytes from prehypertrophic to hypertrophic phenotypes followed by matrix mineralization resulted in suppression of mineral growth without significant effects on alkaline phosphatase levels or cellular morphology in the cultures. Similarly, addition of the synthetic peptide during the late culture period with the advance of cartilage mineralization suppressed mineral growth in pelleted cultures. These data indicate an important role of interactions of chondrocytes with RGD-containing cartilage matrix proteins through integrins in the regulation of epiphyseal chondrocyte differentiation in pelleted cultures.