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PLoS One ; 10(9): e0137186, 2015.
Artículo en Inglés | MEDLINE | ID: mdl-26334533

RESUMEN

PURPOSE: To evaluate the ocular surface change and the inflammatory response in a rabbit model of short-term exposure keratopathy. METHODS: Short term exposure keratopathy by continuous eyelid opening was induced in New Zealand white rabbits for up to 4 hours. Ultrasound pachymetry was used to detect central total corneal thickness. In vivo confocal microscopy and impression cytology were performed to evaluate the morphology of ocular surface epithelium and the infiltration of inflammatory cells. Immunohistochemistry for macrophage,neutrophil, CD4(+) T cells, and CD8(+) T cells were performed to classify the inflammatory cells. Scanning electron microscopy(SEM) was performed to detect ocular surface change.The concentrations of IL-8, IL-17, Line and TNF-αwere analyzed by multiplex immunobead assay. TUNEL staining was performed to detect cellular apoptosis. RESULTS: Significant decrease ofcentral total cornealthickness were found within the first 5 minutes and remained stable thereafter, while there were no changes of corneal epithelial thickness.No significant change of corneal, limbal and conjunctival epithelial morphology was found by in vivo confocal microscopy except the time dependent increase of superficial cellular defects in the central cornea. Impression cytology also demonstrated time dependent increase of sloughing superficial cells of the central cornea. Aggregations ofinflammatory cells were found at 1 hour in the limbal epithelium, 2 hours in the perilimbal conjunctival epithelium, and 3 hours in the peripheral corneal epithelium.In eyes receiving exposure for 4 hours, the infiltration of the inflammatory cells can still be detected at 8 hours after closing eyes.Immunohistochemical study demonstrated the cells to be macrophages, neutrophils, CD4-T cells and CD-8 T cells.SEM demonstrated time-depending increase of intercellular border and sloughing of superficial epithelial cells in corneal surface. Time dependent increase of IL-8, IL-17 and TNF-α in tear was found.TUNEL staining revealed some apoptotic cells in the corneal epithelium and superficial stroma at 3 hours after exposure. CONCLUSIONS: Short term exposure keratopathy can cause significant changes to the ocular surface and inflammatory response. Decrease of central total corneal thickness, aggregation of inflammatory cells, and cornea epithelial cell and superficial keratocyte apoptosis were found no less than 4 hours following the insult.


Asunto(s)
Córnea/patología , Queratoconjuntivitis Seca/patología , Animales , Apoptosis , Parpadeo , Linfocitos T CD4-Positivos/patología , Linfocitos T CD8-positivos/patología , Queratocitos de la Córnea/química , Queratocitos de la Córnea/ultraestructura , Desecación , Modelos Animales de Enfermedad , Epitelio/química , Epitelio/patología , Femenino , Inflamación , Interleucina-17/análisis , Interleucina-8/análisis , Limbo de la Córnea/química , Limbo de la Córnea/ultraestructura , Macrófagos/patología , Microscopía Confocal , Microscopía Electrónica de Rastreo , Neutrófilos/patología , Conejos , Propiedades de Superficie , Factor de Necrosis Tumoral alfa/análisis
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