Immunofluorescent Study on Basement Membrane Proteins and Epithelial Survival Following Brief Ethanol Exposure Like Lasek

Abstract

Abstract: : Purpose:To determine the viability of cultured human corneal epithelial cells and the anatomical cleavage plane after 20 sec exposure to 20% ethanol to create an epithelial flap in a manner similar to LASEK. Methods:Tryptan blue staining was used to determine the viability of cultured corneal cultured cells. Immunofluorescent staining using monoclonal antibodies against laminin 5 and integrins α3ß1 and α6ß4 was performed to determine the anatomical location of the cleavage plane in an epithelial flap created in donor human eyes. Results:There was no difference in cell viability between the exposed and non-exposed cultured human corneal epithelial cells (p=0.422), indicating that a brief exposure to ethanol was not toxic. Immnunofluorescence staining revealed positive patchy staining for laminin 5 and integrins in both the lifted flap and the remaining corneal basement membrane, indicating that the cleavage plane was within the corneal basement membrane. Conclusion:Brief exposure to ethanol does not produce immediate death of corneal epithelial cells. The cleavage plane of a corneal epithelial flap located within the epithelial basement membrane is important to the understanding of LASEK in refractive surgery.

Keywords: 372 cornea: epithelium • 549 refractive surgery: other technologies