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I. Song, C. Joo; Flap Reattachment of Rat Corneal Epithelium After 20% Ethanol Exposure . Invest. Ophthalmol. Vis. Sci. 2003;44(13):3841.
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Purpose: To study morphological changes of rat corneal epithelium, basement membrane and stromal keratocytes and flap reattachment followed by 20% ethanol-exposed flap. Methods: Sprague-Dawley Rats were anesthetized and the central cornea was trephined (2mm in diameter) and exposed (3mm in diameter) to 20% ethanol for 30 seconds. In Group I, corneal epithelial flap was lifted and then repositioned and in Group II, cornea was not treated additionally. The eyes were enucleated, fixed and processed for light microscopy and transmission electron microscopy at indicated time points (0hr, 4hrs, 8hrs, 12hrs, 1day, 3days and 7 days). TUNEL assay and PCNA stain were performed to detect apoptotic cell death and proliferation of ethanol-mediated epithelial flap. For identification of flap attachment, Y chromosome specific gene (SRY) was detected in genomic DNA obtained from female corneal epithelial cells with male epithelial flap using reverse transcription polymerase chain reaction. Results: Hemidesmosomes were not found and the majority of basal cells were damaged in Group I at day1 after reposition. In addition, a space between the basement membrane layer and epithelial flap was observed. Meanwhile ultrastructure of the basement membrane and the hemidesmosomes in basal layer were intact in Group II. Apoptotic cell death and proliferation of basal cells were significantly observed in Group I, while minimal increase was observed in Group II. SRY gene was detected in female corneal epithelial cells with male flap at day1, suggesting that reattachment of ethanol-exposed epithelial flap occurred. Conclusions: Although 20% ethanol exposure for 30 sec may induce some apoptosis of stromal keatocytes and corneal epithelial cell in the edge, it does not give a significant damage to the corneal epithelium. In contrast 20% ethanol-mediated epithelial flap caused a severe damage of the basal cell layer. However, some basal cells were still vital and proliferated and this damage was rapidly recovered.
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