Enzymatic and oxidative stresses are implicated in cartilage degradation occurring in most joint diseases. The aim of this study was to investigate MMP-2, TIMP-1 and -2 activities and F2-Isoprostane concentration in supernatants of cultured chondrocytes (from ponies supplemented with glucosamine and chondroitin sulphate (G/CS) or with placebo) with or without IL-1-beta stimulation. Ponies were orally supplemented during 6 weeks with G/CS (group A) or a placebo (group B). They were then euthanised for teaching purposes allowing us to sample cartilage. Chondrocytes were cultured with or without IL-1-beta stimulation during 6 or 24 h. MMP-2, TIMP-1 and -2 were determined in cell supernatant by zymography and reverse zymography while F2-Isoprostanes concentration was determined by EIA kit (Cayman). Results for MMP-2, TIMP-1 and -2 showed an increasing profile after IL-1-beta stimulation particularly in group B (placebo) although it did not reach significance except for MMP-2 after 24h stimulation (p=0.43, 0.63 and 0.09 respectively for MMP-2, TIMP-1 and TIMP-2 after 6h stimulation and 0.01, 0.36 and 0.30 respectively after 24h stimulation) . A significant time effect was observed, MMP-2, TIMP-1 and -2 activities being higher after 24h IL-1-beta stimulation than after 6h (p<0.05). Differences between group A and B were not significant. In non-stimulated chondrocytes, F2-Isoprostanes concentration was lower in group A than in group B (p<0.05). After IL-1-beta stimulation during 24h, F-2-Isoprostanes increased in both groups but concentrations of group A remained lower than in group B (p<0.05). In conclusion we can say that oral supplementation with glucosamine and chondroitin sulphate can affect F2-Isoprostanes concentration in IL-1-beta stimulated equine chondrocytes and could perhaps affect MMP-2, TIMP-1 and -2 activities but this aspect should be more investigated in further studies.