Effect of sulfate concentration on glycosaminoglycan synthesis in explant cultures of bovine articular cartilage

Abstract

The effects of the sulfate- and FCS concentration on the rate of synthesis and the biochemical properties of glycosaminoglycans, synthesized in bovine articular cartilage in vitro, were studied. 20% FCS in the culture medium stimulated the rate of synthesis. In media without FCS, the rate of synthesis decends from day 0 on. The differences in incorporation rates of [35S]-sodium sulfate and 1,6-[3H]-glucosamine-HCl into glycosaminoglycans in serum free media containing 9 microM and 900 microM sulfate were used to discuss the inorganic sulfate concentration in cartilage. In 9 microM sulfate medium, the newly synthesized glycosaminoglycans contain higher levels of unsulfated disaccharides than the endogenous glycosaminoglycans. In each culture medium, the ratio 6-sulfated disaccharides to 4-sulfated disaccharides of the newly synthesized glycosaminoglycans becomes higher after 3 days in culture. The glycosaminoglycan synthesis is underestimated, when chondrocytes are cultured in media containing less than 200 microM sulfate.