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Effects of triamcinolone acetonide, sodium hyaluronate, amikacin sulfate, and mepivacaine hydrochloride, alone and in combination, on morphology and matrix composition of lipopolysaccharide-challenged and unchallenged equine articular cartilage explants

David M. Bolt Dr med vet, MS1, Akikazu Ishihara BVSc2, Stephen E. Weisbrode VMD, PhD3, and Alicia L. Bertone DVM, PhD4
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  • 1 Comparative Orthopaedic Research Laboratories, Department of Veterinary Clinical Sciences, The Ohio State University, Columbus, OH 43210.
  • | 2 Comparative Orthopaedic Research Laboratories, Department of Veterinary Biosciences, The Ohio State University, Columbus, OH 43210.
  • | 3 Comparative Orthopaedic Research Laboratories, Department of Veterinary Biosciences, The Ohio State University, Columbus, OH 43210.
  • | 4 Comparative Orthopaedic Research Laboratories, Department of Veterinary Clinical Sciences, The Ohio State University, Columbus, OH 43210.

Abstract

Objective—To evaluate the effects of triamcinolone acetonide (TA), sodium hyaluronate (HA), amikacin sulfate (AS), and mepivacaine hydrochloride (MC) on articular cartilage morphology and matrix composition in lipopolysaccharide (LPS)-challenged and unchallenged equine articular cartilage explants.

Sample Population—96 articular cartilage explants from 4 femoropatellar joints of 2 adult horses.

Procedures—Articular cartilage explants were challenged with LPS (100 ng/mL) or unchallenged for 48 hours, then treated with TA, HA, AS, and MC alone or in combination for 96 hours or left untreated. Cartilage extracts were analyzed for glycosaminoglycan (GAG) content by dimethyl-methylene blue assay (ng/mg of dry wt). Histomorphometric quantification of total lacunae, empty lacunae, and lacunae with pyknotic nuclei was recorded for superficial, middle, and deep cartilage zones.

Results—LPS induced a significant increase in pyknotic nuclei and empty lacunae. Treatment with TA or HA significantly decreased empty lacunae (TA and HA), compared with groups without TA or HA, and significantly decreased empty lacunae of LPS-challenged explants, compared with untreated explants. Treatment with AS or MC significantly increased empty lacunae in unchallenged explants, and these effects were attenuated by TA. Treatment with MC significantly increased empty lacunae and pyknotic nuclei and, in combination with LPS, could not be attenuated by TA. Content of GAG did not differ between unchallenged and LPS-challenged explants or among treatments.

Conclusions and Clinical Relevance—Treatment with TA or HA supported chondrocyte morphology in culture and protected chondrocytes from toxic effects exerted by LPS, AS, and MC.

Contributor Notes

Dr. Bolt's present address is Department of Veterinary Clinical Sciences, Royal Veterinary College, University of London, Hatfield, Hertfordshire AL9 7TA, England.

The authors thank David Spencer Smith and Dr. Kelly Santangelo for technical assistance.

Address correspondence to Dr. Bertone.