C E Kawcak1, G W Trotter, D D Frisbie, C W McIlwraith. 1. Department of Clinical Sciences, College of Veterinary Medicine and Biomedical Sciences, Colorado State University, Fort Collins 80523, USA.
Abstract
OBJECTIVE: To evaluate the effects of a commercially defined, serum-free medium additive on equine articular cartilage explants, compared with effects of serum-free and serum-supplemented media. ANIMALS: Articular cartilage from a 3-year-old, mixed breed horse euthanatized for reasons other than musculoskeletal disease or sepsis. PROCEDURE: Media were changed every 48 hours, and the glycosaminoglycan (GAG) content was determined in media collected at each time point. Glycosaminoglycan synthesis by explant chondrocytes, and residual GAG content of articular cartilage (as a measure of explant GAG loss) were determined at the end of the study (day 8). RESULTS: Articular cartilage explants in serum-free medium and the commercial supplemented medium had significantly lower GAG synthesis and GAG content than did those incubated in serum-supplemented medium. There were no significant differences in GAG synthesis and content between serum-free and commercial supplemented medium groups. When comparing medium GAG content for all treatment groups, the GAG content in serum-free medium on day 8 was significantly greater than that in commercial supplemented medium, but significant differences were not evident in percentage of release of GAG (as an indicator of GAG degradation) among all 3 treatment groups. CONCLUSIONS: Commercial supplemented medium had effects on articular cartilage matrix GAG loss into medium equal to those of serum-supplemented medium (eg, both lost articular cartilage explant GAG to a similar degree). However, residual articular cartilage GAG content was higher in serum-supplemented medium, as was GAG synthesis. Commercial supplemented medium appears to either lack the proper ingredients to maintain steady-state GAG synthesis, or lacks proper concentrations of these ingredients.
OBJECTIVE: To evaluate the effects of a commercially defined, serum-free medium additive on equinearticular cartilage explants, compared with effects of serum-free and serum-supplemented media. ANIMALS: Articular cartilage from a 3-year-old, mixed breed horse euthanatized for reasons other than musculoskeletal disease or sepsis. PROCEDURE: Media were changed every 48 hours, and the glycosaminoglycan (GAG) content was determined in media collected at each time point. Glycosaminoglycan synthesis by explant chondrocytes, and residual GAG content of articular cartilage (as a measure of explant GAG loss) were determined at the end of the study (day 8). RESULTS:Articular cartilage explants in serum-free medium and the commercial supplemented medium had significantly lower GAG synthesis and GAG content than did those incubated in serum-supplemented medium. There were no significant differences in GAG synthesis and content between serum-free and commercial supplemented medium groups. When comparing medium GAG content for all treatment groups, the GAG content in serum-free medium on day 8 was significantly greater than that in commercial supplemented medium, but significant differences were not evident in percentage of release of GAG (as an indicator of GAG degradation) among all 3 treatment groups. CONCLUSIONS: Commercial supplemented medium had effects on articular cartilage matrix GAG loss into medium equal to those of serum-supplemented medium (eg, both lost articular cartilage explant GAG to a similar degree). However, residual articular cartilageGAG content was higher in serum-supplemented medium, as was GAG synthesis. Commercial supplemented medium appears to either lack the proper ingredients to maintain steady-state GAG synthesis, or lacks proper concentrations of these ingredients.