Maintenance of equine articular cartilage explants in serum-free and serum-supplemented media, compared with that in a commercial supplemented medium

Christopher E. Kawcak From the Orthopedic Research Laboratory, Department of Clinical Sciences, College of Veterinary Medicine and Biomedical Sciences, Colorado State University, Fort Collins, CO 80523.

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Gayle W. Trotter From the Orthopedic Research Laboratory, Department of Clinical Sciences, College of Veterinary Medicine and Biomedical Sciences, Colorado State University, Fort Collins, CO 80523.

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David D. Frisbie From the Orthopedic Research Laboratory, Department of Clinical Sciences, College of Veterinary Medicine and Biomedical Sciences, Colorado State University, Fort Collins, CO 80523.

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C. Wayne McIlwraith From the Orthopedic Research Laboratory, Department of Clinical Sciences, College of Veterinary Medicine and Biomedical Sciences, Colorado State University, Fort Collins, CO 80523.

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 BVSc, PhD

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 steadystate GAG synthesis, or lacks proper concentrations of these ingredients. (Am J Vet Res 1996;57:1261–1265)

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 steadystate GAG synthesis, or lacks proper concentrations of these ingredients. (Am J Vet Res 1996;57:1261–1265)

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