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Rangan Gupta1,2, Benjamin W. Yarnall1,3, Elizabeth A. Giuliano1,3, Jagat R. Kanwar4, Dylan G. Buss1,3, Rajiv R. Mohan1,2,3
Article first published online: 18 APR 2011
© 2011 American College of Veterinary Ophthalmologists
Keywords: canine; cornea; fibroblasts; mitomycin C
Objective To evaluate the safety and efficacy of mitomycin C (MMC) in prevention of canine corneal scarring.
Methods With an in vitro approach using healthy canine corneas, cultures of primary canine corneal fibroblasts or myofibroblasts were generated. Primary canine corneal fibroblasts were obtained by growing corneal buttons in minimal essential medium supplemented with 10% fetal bovine serum. Canine corneal myofibroblasts were produced by growing cultures in serum-free medium containing transforming growth factor β1 (1 ng/mL). Trypan blue assay and phase-contrast microscopy were used to evaluate the toxicity of three doses of MMC (0.002%, 0.02% and 0.04%). Real-time PCR, immunoblot, and immunocytochemistry techniques were used to determine MMC efficacy to inhibit markers of canine corneal scarring.
Results A single 2-min treatment of 0.02% or less MMC did not alter canine corneal fibroblast or keratocyte phenotype, viability, or growth. The 0.02% dose substantially reduced myofibroblast formation (up to 67%; P < 0.001), as measured by the change in RNA and protein expression of fibrosis biomarkers (α-smooth muscle actin and F-actin).
Conclusion This in vitro study suggests that a single 2-min 0.02% MMC treatment to the canine corneal keratocytes is safe and may be useful in decreasing canine corneal fibrous metaplasia. In vivo studies are warranted.
View the full article from Veterinary Ophthalmology Vol. 14 Issue 3 in HTML or PDF
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