Live or Let Die: Epithelial Flap Vitality and Keratocyte Proliferation Following LASEK and Epi-LASIK in Human Donor and Porcine Eyes
Romesh I. Angunawela, C. Winkler von Mohrenfels, Anupma Kumar, David O’Brart, John Marshall
Abstract
Romesh I. Angunawela, C. Winkler von Mohrenfels, Anupma Kumar, David O’Brart, John Marshall
Abstract
PURPOSE: To determine the relationship between epithelial flap vitality and stromal keratocyte proliferation following two epithelial refractive techniques: epi-LASIK and laser epithelial keratomileusis (LASEK). METHODS: Human corneas were maintained in organ culture and underwent standard -6.00-diopter ablation. Rates of stromal keratocyte proliferation were detected 1 week postoperative using a Ki67 antibody specific to proliferating cells. Images were captured with a laser scanning confocal microscope and analyzed by a masked observer. Epithelial flap vitality was determined with propidium iodide using fresh porcine corneas. Epithelial flaps were created with Gebauer Epikeratome epi-LASIK or alcohol-assisted LASEK method. Flaps treated with 100% alcohol and uninjured corneas were used as controls. RESULTS: The number of proliferating keratocytes was greatest at 1 week in the epi-LASIK corneas (P<.001). Cell vitality was greatest in the epi-LASIK flaps and declined in the LASEK and 100% alcohol flaps (P<.001). CONCLUSIONS: In this in vitro setting, epi-LASIK results in an epithelial flap with significantly more live cells. There is also a greater number of proliferating stromal cells following epi-LASIK at 1 week. Based on these in vitro observations, epi-LASIK may result in greater levels of haze compared to LASEK.
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PURPOSE: To determine the relationship between epithelial flap vitality and stromal keratocyte proliferation following two epithelial refractive techniques: epi-LASIK and laser epithelial keratomileusis (LASEK). METHODS: Human corneas were maintained in organ culture and underwent standard -6.00-diopter ablation. Rates of stromal keratocyte proliferation were detected 1 week postoperative using a Ki67 antibody specific to proliferating cells. Images were captured with a laser scanning confocal microscope and analyzed by a masked observer. Epithelial flap vitality was determined with propidium iodide using fresh porcine corneas. Epithelial flaps were created with Gebauer Epikeratome epi-LASIK or alcohol-assisted LASEK method. Flaps treated with 100% alcohol and uninjured corneas were used as controls. RESULTS: The number of proliferating keratocytes was greatest at 1 week in the epi-LASIK corneas (P<.001). Cell vitality was greatest in the epi-LASIK flaps and declined in the LASEK and 100% alcohol flaps (P<.001). CONCLUSIONS: In this in vitro setting, epi-LASIK results in an epithelial flap with significantly more live cells. There is also a greater number of proliferating stromal cells following epi-LASIK at 1 week. Based on these in vitro observations, epi-LASIK may result in greater levels of haze compared to LASEK.
Key concepts: Keratomileusis, Medicine, LASIK, Stromal cell, Dioptre, Ophthalmology, Surgery, Cornea