Purchase this article with an account.
R. Cantalapiedra, T. Blanco, J. Hincapie, P. Perez-Merino, S. Mar, L. Ibares-Frias, P. Gallego, S. del Olmo-Aguado, I. Alcalde, J. Merayo-Lloves; Collagen Cross-Linking Procedure Induces Severe Damage in Corneas Without Bowman’s Layer, in Hens. Invest. Ophthalmol. Vis. Sci. 2009;50(13):914.
Download citation file:
© ARVO (1962-2015); The Authors (2016-present)
In a 6,00 mm diameter area, the Bowman layer of both eyes of 28 hens was removed with an excimer laser. After two months the right corneas, were UVA-irradiated (360-380 nm, 3 mW/cm2), in a 6,00 mm diameter deepithelized area, during 30 minutes. A 0,1% riboflavin-20% dextran solution was administered 5 minutes before irradiation and then every 5 minutes during the UVA irradiation. The left fellow control corneas were only debridement. Then, animals were allowed to heal for up to 1 month and euthanized in different after 6, 12 and 24 hours, 3, 5, 15 and 30 days, being the corneas fixed for pathology. Cell death was evaluated with TUNEL assay. Anti BrdU for cell proliferation and expression of -SMA indicative of myofibroblast differentiation were evaluated by indirect immunofluorescence microscopy.
Differences in the time of epithelial closure (48±8 hours) were not observed although a great grade of edema was observed in the CXL corneas (p<0.05). The corneal thickness remains higher in the right corneas (p<0.05). Endothelial damage was observed in 100% of CXL corneas and in none of the control. Although subepithelial haze was not obserbed in any cornea, endothelial haze was observed in 100% of right corneas. Presence of inflammatory response was observed higher in the anterior chamber of irradiated eyes (p<0,05).Keratocytes and endothelial cells were full disappeared in the irradiated area in the CXL corneas and only in the anterior stromal third in the controls. Proliferation of new endothelial cells and keratocytes was observed at the third day after treatment. At the fifth day presence of myofibroblast was observed closing the endothelial holds. At the fifteenth day the endothelium was full restored again with new endothelial cells. At day thirtieth all the corneas appears with a normal cell density and subepithelial fibrosis was not seen.
This PDF is available to Subscribers Only