Effects of glucosamine and chondroitin sulfate on mediators of osteoarthritis in cultured equine chondrocytes stimulated by use of recombinant equine interleukin-1β

Kirsten M. Neil Department of Large Animal Clinical Sciences, College of Veterinary Medicine, Michigan State University, East Lansing, MI 48824.
Present address is Goulburn Valley Equine Hospital, 905 Goulburn Valley Hwy, Congupna Victoria 3633, Australia.

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Michael W. Orth Department of Animal Science, College of Agriculture and Natural Resources, Michigan State University, East Lansing, MI 48824.

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Paul M. Coussens Department of Animal Science, College of Agriculture and Natural Resources, Michigan State University, East Lansing, MI 48824.

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Pooi-See Chan Department of Animal Science, College of Agriculture and Natural Resources, Michigan State University, East Lansing, MI 48824.

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John P. Caron Department of Large Animal Clinical Sciences, College of Veterinary Medicine, Michigan State University, East Lansing, MI 48824.

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Abstract

Objective—To determine whether glucosamine and chondroitin sulfate (CS) at concentrations approximating those achieved in plasma by oral administration would influence gene expression of selected mediators of osteoarthritis in cytokine-stimulated equine articular chondrocytes.

Sample Population—Samples of grossly normal articular cartilage obtained from the metacarpophalangeal joint of 13 horses.

Procedure—Equine chondrocytes in pellet culture were stimulated with a subsaturating dose of recombinant equine interleukin (reIL)-1β. Effects of prior incubation with glucosamine (2.5 to 10.0 µg/mL) and CS (5.0 to 50.0 µg/mL) on gene expression of matrix metalloproteinase (MMP)-1, -2, -3, -9, and -13; aggrecanase 1 and 2; inducible nitric oxide synthase (iNOS); cyclooxygenase (COX)-2; nuclear factor κB; and c-Jun- N-terminal kinase (JNK) were assessed by use of a quantitative real-time polymerase chain reaction assay.

Results—Glucosamine at a concentration of 10 µg/mL significantly reduced reIL-1β–induced mRNA expression of MMP-13, aggrecanase 1, and JNK. Reductions in cytokine-induced expression were also observed for iNOS and COX-2. Chondroitin sulfate had no effect on gene expression at the concentrations tested.

Conclusions and Clinical Relevance—Concentrations of glucosamine similar to those achieved in plasma after oral administration in horses exerted pretranslational regulation of some mediators of osteoarthritis, an effect that may contribute to the cartilage- sparing properties of this aminomonosaccharide. Analysis of results of this study indicated that the influence of CS on pretranslational regulation of these selected genes is limited or lacking. (Am J Vet Res 2005;66:1861–1869)

Abstract

Objective—To determine whether glucosamine and chondroitin sulfate (CS) at concentrations approximating those achieved in plasma by oral administration would influence gene expression of selected mediators of osteoarthritis in cytokine-stimulated equine articular chondrocytes.

Sample Population—Samples of grossly normal articular cartilage obtained from the metacarpophalangeal joint of 13 horses.

Procedure—Equine chondrocytes in pellet culture were stimulated with a subsaturating dose of recombinant equine interleukin (reIL)-1β. Effects of prior incubation with glucosamine (2.5 to 10.0 µg/mL) and CS (5.0 to 50.0 µg/mL) on gene expression of matrix metalloproteinase (MMP)-1, -2, -3, -9, and -13; aggrecanase 1 and 2; inducible nitric oxide synthase (iNOS); cyclooxygenase (COX)-2; nuclear factor κB; and c-Jun- N-terminal kinase (JNK) were assessed by use of a quantitative real-time polymerase chain reaction assay.

Results—Glucosamine at a concentration of 10 µg/mL significantly reduced reIL-1β–induced mRNA expression of MMP-13, aggrecanase 1, and JNK. Reductions in cytokine-induced expression were also observed for iNOS and COX-2. Chondroitin sulfate had no effect on gene expression at the concentrations tested.

Conclusions and Clinical Relevance—Concentrations of glucosamine similar to those achieved in plasma after oral administration in horses exerted pretranslational regulation of some mediators of osteoarthritis, an effect that may contribute to the cartilage- sparing properties of this aminomonosaccharide. Analysis of results of this study indicated that the influence of CS on pretranslational regulation of these selected genes is limited or lacking. (Am J Vet Res 2005;66:1861–1869)

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