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Asadolah Movahedan, Neda Afshar, Medi Eslani, Sanaz Gidfar, Ali R Djalilian; The effect of regional limbal injury on mouse corneal epithelium. Invest. Ophthalmol. Vis. Sci. 2014;55(13):3123. doi: https://doi.org/.
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To define the effect of localized limbal injury in the development of regional limbal epithelial dysfunction in the mouse cornea
Adult 4-months-old C57BL6J mice were used to develop different models of limbal dysfunction according to the statement for the use of animals in Association for Research in Vision and Ophthalmology. Limbal injury was inflicted by corneal limbal epithelium ablation using Alger brush or fine electrode thermal cauterization in different limbal locations (superior nasal, superior temporal, inferior nasal, inferior temporal quadrants, upper, lower, temporal and nasal halves). Whole (360 degrees) limbal ablation was also performed. Slit-lamp examination was used to detect late fluorescein surface staining. Whole mount Immunofluorescent staining for Cytokeratins 12 and 8 were used to evaluate corneal surface cell types 2 months after the injuries.
Limbal injury in quadrants (90 degrees) did not cause any epitheliopathy (late fluorescein staining) in any location (superior nasal, superior temporal, inferior nasal or inferior temporal) in slit-lamp examination or whole mount staining compared to unwounded control 2 months after the injury. In contrast 180 degrees of limbal injury resulted in a mild epitheliopathy in the corresponding half of cornea. 360 degrees of limbal ablation using Alger brush resulted in partial limbal stem cell deficiency which was more severe with neovascularization and a more complete loss of Cytokeratin12 if an additional 360 degree round of ablation was performed at the time injury.
At least a 180 degrees of limabl injury is required in mouse model of limbal stem cell deficiency for development of epitheliopathy manifested by late fluorescein staining 2 months after the injury. By increasing the severity of limbal injury by performing an additional round of 360 degrees of mechanical limbal ablation a more complete model of limbal stem cell deficiency with severe neovascularization can be achieved.
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