Effects of [3H]glucosamine concentration on [3H]chondroitin sulphate formation by cultured chondrocytes

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Abstract
GlcN (glucosamine) is now promoted over the counter for implied treatment of osteoarthritis, ostensibly by stimulating biosynthesis of cartilage chondroitin sulphate. In order to evaluate whether exogenous GlcN has any stimulatory effect, we have incubated mouse chondrocytes with [35S]sulphate and various amounts of GlcN, to determine whether any increment in chondroitin [35S]sulphate formation occurs. Similarly we have used varying concentrations of [3H]GlcN to determine the dilution of incorporation into [3H]chondroitin sulphate due to provision of endogenous GlcN by metabolism from glucose at two different glucose concentrations. The incorporation of both 35S and 3H was essentially linear over a 5 h time period. We found no stimulation of chondroitin [35S]sulphate synthesis at lower concentrations of GlcN, and a significant reduction at higher concentrations. Even at concentrations of [3H]GlcN that were greater than could be achieved with standard doses of oral GlcN, there was significant dilution of exogenous GlcN. Furthermore, an artificial acceptor for glycosaminoglycan synthesis in cell culture, 4-methylumbelliferyl β-d-xyloside, did not modify the provision of GlcN from endogenous sources, even though it stimulated chondroitin sulphate synthesis 4 –5-fold at each GlcN concentration. We conclude that the cells have excess capacity to form maximal amounts of GlcN from glucose so that exogenous GlcN does not stimulate chondroitin sulphate synthesis.

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