High doses of glucosamine-HCl have detrimental effects on bovine articular cartilage explants cultured in vitro

Objective: To investigate both the biochemical and the potential morphological changes in bovine cartilage explants following treatment with glucosamine HCl, and to evaluate the capability of glucosamine to counteract the degradation of cartilage induced by catabolic agents such as interleukin-1beta (IL-1beta) and the bacterial lipopolysaccharide (LPS). Design: Bovine articular cartilage explants were treated with increasing doses of glucosamine HCl (0.25-25 mg/ml) in the absence or in the presence of IL-1beta or LPS. The release of matrix proteoglycans in the medium, as well as variations in nitric oxide and lactate production were evaluated by standard assays. Proteoglycan synthesis was determined by incorporation of (Na2SO4)-S-35. Ultrastructural analysis was performed by transmission electron microscopy. Results: Increasing doses of glucosamine (2.5, 6.5, 25 mg/ml) induced a dose-dependent decrease in proteoglycan synthesis and in lactate production after 24 h treatment. The biochemical changes induced by IL1-beta or LPS appeared to be inhibited by 6.5 and 25 mg/ml glucosamine. At these concentrations a decrease in cell viability was observed, which reached over 90% at 25 mg/ml. Conclusions: This study shows that pharmacological doses of glucosamine induce a broad impairment in the metabolic activity of bovine chondrocytes, leading to cell death. The inhibition of the catabolic effects induced by IL1-beta and LPS appears related to glucosamine toxicity. In other experimental models, the same or similar doses of glucosamine have previously been used, without showing any adverse effect. We conclude that, in studying the effects of glucosamine, particular attention should be addressed to the experimental model, the doses and the length of treatment. (C) 2002 OsteoArthritis Research Society International. Published by Elsevier Science Ltd. All rights reserved.