Evaluation of species differences and the effects of storage duration and temperature on the anticollagenase efficacy of canine, feline, and equine serum on in vitro corneal degradation

Emily D. Conway Department of Veterinary Clinical Sciences, College of Veterinary Medicine, Purdue University, West Lafayette, IN 47907.

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 BVMS, MS
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Jean Stiles Department of Veterinary Clinical Sciences, College of Veterinary Medicine, Purdue University, West Lafayette, IN 47907.

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 DVM, MS
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Wendy M. Townsend Department of Veterinary Clinical Sciences, College of Veterinary Medicine, Purdue University, West Lafayette, IN 47907.

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Hsin-Yi Weng Department of Comparative Pathobiology, College of Veterinary Medicine, Purdue University, West Lafayette, IN 47907.

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 BVM, MPH, PhD

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Abstract

OBJECTIVE To evaluate species differences and effects of storage duration and temperature on the anticollagenase efficacy of canine, feline, and equine serum on in vitro corneal degradation.

SAMPLES Corneas and serum from dogs, cats, and horses.

PROCEDURES Clinically normal corneas from dogs, cats, and horses were harvested within 2 hours after euthanasia. Serum samples from dogs, cats, and horses were collected and pooled by species. Corneal specimens were incubated with collagenase derived from Clostridium histolyticum, 5mM calcium chloride in saline (0.9% NaCl) solution, and feline, canine, or equine serum that had been stored for 0, 30, 90, or 180 days at −20° or −80°C. Following incubation, the corneal weight loss percentage and hydroxyproline concentration in the incubation fluid were calculated and compared among experimental combinations.

RESULTS Feline serum was more effective than canine or equine serum for minimizing corneal weight loss. Incubation with feline or equine, but not canine, serum significantly reduced hydroxyproline production. Serum storage duration did not affect corneal weight loss, but the hydroxyproline concentration was greater for corneal specimens that were incubated with serum that was stored for 90 days, compared with that for corneal specimens incubated with serum that was stored for 0, 30, or 180 days. Serum storage temperature did not affect corneal weight loss or hydroxyproline concentration.

CONCLUSIONS AND CLINICAL RELEVANCE Results suggested that serum reduced corneal degradation in vitro, and the duration and temperature at which serum was stored did not affect its anticollagenase efficacy.

Abstract

OBJECTIVE To evaluate species differences and effects of storage duration and temperature on the anticollagenase efficacy of canine, feline, and equine serum on in vitro corneal degradation.

SAMPLES Corneas and serum from dogs, cats, and horses.

PROCEDURES Clinically normal corneas from dogs, cats, and horses were harvested within 2 hours after euthanasia. Serum samples from dogs, cats, and horses were collected and pooled by species. Corneal specimens were incubated with collagenase derived from Clostridium histolyticum, 5mM calcium chloride in saline (0.9% NaCl) solution, and feline, canine, or equine serum that had been stored for 0, 30, 90, or 180 days at −20° or −80°C. Following incubation, the corneal weight loss percentage and hydroxyproline concentration in the incubation fluid were calculated and compared among experimental combinations.

RESULTS Feline serum was more effective than canine or equine serum for minimizing corneal weight loss. Incubation with feline or equine, but not canine, serum significantly reduced hydroxyproline production. Serum storage duration did not affect corneal weight loss, but the hydroxyproline concentration was greater for corneal specimens that were incubated with serum that was stored for 90 days, compared with that for corneal specimens incubated with serum that was stored for 0, 30, or 180 days. Serum storage temperature did not affect corneal weight loss or hydroxyproline concentration.

CONCLUSIONS AND CLINICAL RELEVANCE Results suggested that serum reduced corneal degradation in vitro, and the duration and temperature at which serum was stored did not affect its anticollagenase efficacy.

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