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1From the Evelyn F. and William L. McKnight Vision Research Center, Bascom Palmer Eye Institute, University of Miami School of Medicine, Miami Florida; the 2Schepens Eye Research Institute and Departments of Ophthalmology and Cell Biology, Harvard Medical School, Boston Massachusetts; and the 3Department of Biology, College of Natural Sciences, Kyungpook National University, Taegu, Korea.
PURPOSE. Incisional or ablation injury to the corneal stroma is repaired by deposition of a fibrotic tissue produced by activated keratocytes, whereas cells lost from the underlying stroma after epithelial abrasion are simply replaced by keratocyte replication without expression of fibrotic markers. The purpose of this study was to investigate mechanisms that determine this differential keratocyte response.
METHODS. A penetrating keratectomy rabbit model was adapted for mice to study the fibrotic repair response. A mouse epithelial abrasion model was applied to study the stromal cell replacement response. A primary rabbit corneal cell culture model and an organotypic culture model were also used.
RESULTS. When the epithelium was prevented from resurfacing the cornea after penetrating keratectomy, expression of fibrotic markers was considerably reduced. TGF-ß2 was determined to be a major substance produced by corneal epithelial cells capable of inducing the fibrotic phenotype. In the intact mouse cornea, TGF-ß2 was confined to the uninjured epithelium, but was released into the stroma during fibrotic repair. By contrast, TGF-ß1 was never found in the epithelium. When epithelial cells were cultured on a basement-membrane–like gel or allowed to deposit their own basement membrane in organotypic culture, TGF-ß2 production was reduced. Return of a basement membrane after wounding in vivo correlated with loss of the fibrotic phenotype. In the epithelial debridement injury model in which the basement membrane was left intact, TGF-ß2 remained confined to the corneal epithelium, consistent with the absence of a fibrotic phenotype.
CONCLUSIONS. These data suggest that integrity of the basement membrane is a deciding factor in determining the regenerative character of corneal repair.
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