Effect of corneal epithelium on ultraviolet-A and riboflavin absorption

PURPOSE: To determine if the corneal epithelium prevents the collagen cross-linking effect. Using immunofluorescence microscopy after CXL, we indirectly analyzed the role of the epithelium as ultraviolet-A (UVA) shield as well as a barrier to riboflavin penetration. METHODS: Fifteen freshly enucleat...

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Detalles Bibliográficos
Autores: Bottós, Katia Mantovani [UNIFESP], Schor, Paulo [UNIFESP], Dreyfuss, Juliana Luporini [UNIFESP], Nader, Helena Bonciani [UNIFESP], Chamon, Wallace [UNIFESP]
Tipo de recurso: artículo
Estado:Versión publicada
Fecha de publicación:2011
País:Brasil
Institución:Universidade Federal de São Paulo (UNIFESP)
Repositorio:Repositório Institucional da UNIFESP
Idioma:inglés
OAI Identifier:oai:repositorio.unifesp.br:11600/6620
Acceso en línea:http://dx.doi.org/10.1590/S0004-27492011000500008
http://repositorio.unifesp.br/handle/11600/6620
Access Level:acceso abierto
Palabra clave:Cross-linking reagents
Riboflavin
Epithelium, corneal
Microscopy, immunofluorescence
Photosensitizing agents
Ultraviolet rays
Swine
Reagentes para ligações cruzadas
Riboflavina
Epitélio corneano
Microscopia de imunofluorescência
Agentes fotossensibilizantes
Raios ultravioleta
Suíno
Descripción
Sumario:PURPOSE: To determine if the corneal epithelium prevents the collagen cross-linking effect. Using immunofluorescence microscopy after CXL, we indirectly analyzed the role of the epithelium as ultraviolet-A (UVA) shield as well as a barrier to riboflavin penetration. METHODS: Fifteen freshly enucleated porcine eyes were divided into 3 groups. The corneal epithelium was kept intact in all groups. Five eyes served as control (Group 1). On group 2, eyes received tetracaine anesthetic drops and topical 0.1% riboflavin solution (10 mg riboflavin-5-phosphate in 10 mL 20% dextran-T-500). On Group 3, riboflavin was injected into the anterior chamber to allow penetration of the drug through the endothelium. Groups 2 and 3 were exposed to UVA (365 nm, 3 mW/cm²) for 30 minutes. Ultra-thin sections (8 µm) of the corneas were stained with anti-collagen type I and DAPI (4,6-diamidino-2-fenilindole dihydrocloride) and analyzed with fluorescence microscopy. RESULTS: Corneas treated with UVA irradiation and intracameral injection of riboflavin (Group 3) showed greater pattern of collagen organization compared to groups 1 (Control) and 2 (riboflavin and tetracaine eye drops). A yellow stromal staining, which represents the riboflavin diffusion into the stroma, was only observed in eyes injected with riboflavin into the anterior chamber. CONCLUSION: Using immunofluorescence microscopy in porcine corneas, we demonstrated that the corneal epithelium reduces the effectiveness of CXL by preventing the penetration of the drug and not by limiting the UVA transmittance. An inadequate intrastromal concentration of riboflavin may impair CXL effect.