In vitro dose-dependent effects of enrofloxacin on equine articular cartilage

Lisa A. Beluche From the Orthopedic Research Laboratory, Departments of Veterinary Clinical Sciences (Beluche, Bertone, Anderson, Kohn) and Veterinary Biosciences (Weisbrode), The Ohio State University, Columbus, OH 43210.

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 DVM, MS
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Alicia L. Bertone From the Orthopedic Research Laboratory, Departments of Veterinary Clinical Sciences (Beluche, Bertone, Anderson, Kohn) and Veterinary Biosciences (Weisbrode), The Ohio State University, Columbus, OH 43210.

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David E. Anderson From the Orthopedic Research Laboratory, Departments of Veterinary Clinical Sciences (Beluche, Bertone, Anderson, Kohn) and Veterinary Biosciences (Weisbrode), The Ohio State University, Columbus, OH 43210.

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Catherine W. Kohn From the Orthopedic Research Laboratory, Departments of Veterinary Clinical Sciences (Beluche, Bertone, Anderson, Kohn) and Veterinary Biosciences (Weisbrode), The Ohio State University, Columbus, OH 43210.

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Steven E. Weisbrode From the Orthopedic Research Laboratory, Departments of Veterinary Clinical Sciences (Beluche, Bertone, Anderson, Kohn) and Veterinary Biosciences (Weisbrode), The Ohio State University, Columbus, OH 43210.

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 VMD, PhD

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Abstract

Objective

To determine whether enrofloxacin has detrimental, dose-dependent effects on equine articular cartilage in vitro.

Animals

Cartilage explants were developed from 6 healthy horses between 0 and 96 months old.

Procedure

Patellar cartilage explants were incubated in 5 concentrations of enrofloxacin (2 μg/ml, 10 μg/ml, 1,000 μg/ml, 10,000 μg/ml, and 50,000 μg/ml) for 72 hours. Proteoglycan synthesis (Na35SO4 incorporation for 24 hours), proteoglycan degradation (Na35SO4 release for 72 hours), endogenous proteoglycan content (dimethylmethlene blue assay), and total protein content were determined. Cartilage explants were evaluated by use of histomorphologic and histomorphometric techniques (toluidine blue stain) for cytologic and matrix characteristics. Quantitative data were analyzed with a one-way ANOVA to compare results among various enrofloxacin concentration groups and the control group. A general linear model was used to determine whether age had an effect.

Result

Proteoglycan synthesis was excellent in control specimens and in specimens incubated in low concentrations of enrofloxacin (2 μg/ml and 10 μg/ml). High concentrations of enrofloxacin (> 1,000 μg/ml) effectively eliminated proteoglycan synthesis regardless of horse age. Proteoglycan degradation at low concentrations (2 μg/ml and 10 μg/ml) was not different than control. High concentrations of enrofloxacin (> 1,000 μg/ml) caused significant degradation. Different concentrations of enrofloxacin did not affect endogenous proteoglycan. High concentrations of enrofloxacin were associated with a significant increase in number of pyknotic nuclei.

Conclusion

Concentrations of enrofloxacin that might be achieved following systemic administration did not suppress chondrocyte metabolism in vitro. High concentrations of enrofloxacin (> 1,000 μg/ml) were toxic to chondrocytes. (Am J Vet Res 1999;60:577–582)

Abstract

Objective

To determine whether enrofloxacin has detrimental, dose-dependent effects on equine articular cartilage in vitro.

Animals

Cartilage explants were developed from 6 healthy horses between 0 and 96 months old.

Procedure

Patellar cartilage explants were incubated in 5 concentrations of enrofloxacin (2 μg/ml, 10 μg/ml, 1,000 μg/ml, 10,000 μg/ml, and 50,000 μg/ml) for 72 hours. Proteoglycan synthesis (Na35SO4 incorporation for 24 hours), proteoglycan degradation (Na35SO4 release for 72 hours), endogenous proteoglycan content (dimethylmethlene blue assay), and total protein content were determined. Cartilage explants were evaluated by use of histomorphologic and histomorphometric techniques (toluidine blue stain) for cytologic and matrix characteristics. Quantitative data were analyzed with a one-way ANOVA to compare results among various enrofloxacin concentration groups and the control group. A general linear model was used to determine whether age had an effect.

Result

Proteoglycan synthesis was excellent in control specimens and in specimens incubated in low concentrations of enrofloxacin (2 μg/ml and 10 μg/ml). High concentrations of enrofloxacin (> 1,000 μg/ml) effectively eliminated proteoglycan synthesis regardless of horse age. Proteoglycan degradation at low concentrations (2 μg/ml and 10 μg/ml) was not different than control. High concentrations of enrofloxacin (> 1,000 μg/ml) caused significant degradation. Different concentrations of enrofloxacin did not affect endogenous proteoglycan. High concentrations of enrofloxacin were associated with a significant increase in number of pyknotic nuclei.

Conclusion

Concentrations of enrofloxacin that might be achieved following systemic administration did not suppress chondrocyte metabolism in vitro. High concentrations of enrofloxacin (> 1,000 μg/ml) were toxic to chondrocytes. (Am J Vet Res 1999;60:577–582)

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