Effects of human recombinant interleukin-1beta on canine articular chondrocytes in three-dimensional culture
- PMID: 10895897
- DOI: 10.2460/ajvr.2000.61.766
Effects of human recombinant interleukin-1beta on canine articular chondrocytes in three-dimensional culture
Abstract
Objective: To determine the effects of interleukin (IL)-1beta on matrix synthesis and degradation by chondrocytes cultured in a 3-dimensional (3-D) gel medium.
Sample population: Chondrocytes from 7 dogs.
Procedure: Articular chondrocytes were harvested and cultured in 3-D gel medium alone or with 10 or 20 ng IL-1beta/ml that was added beginning on day 0, 3, 6, or 9. On days 3, 6, 12, and 20 of 3-D culture, samples of the liquid medium were evaluated for glycosaminoglycan (GAG), prostaglandin E2 (PGE2), and matrix metalloprotease (MMP)-3 content. The 3-D plug in each well was evaluated for histologic characteristics of viability, cell morphology, and proteoglycan staining, immunohistochemically stained for collagen type II, and spectrophotometrically analyzed for GAG content.
Results: Significant differences for all variables were detected between controls and each IL-1beta group, among groups with different IL-1beta concentrations, and among groups with IL-1beta added at various time points. Chondrocytes exposed to IL-1beta had loss of GAG, increased PGE2 and MMP-3 concentrations, and lack of collagen type-II synthesis. These IL-1beta effects appeared to be time and concentration dependent.
Conclusions: Addition of IL-1beta to chondrocytes in 3-D gel medium results in time- and concentration-dependent effects on matrix synthesis and degradation and provides an appropriate in vitro model for many of the pathophysiologic events associated with osteoarthritis.
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