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J. Huang, D.E. Possin, S.E. Wilson; Monocyte Influx Into the Mouse Corneal Stroma Following Epithelial Scrape Injury . Invest. Ophthalmol. Vis. Sci. 2003;44(13):888.
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Purpose: To characterize monocyte infiltration into the mouse cornea after epithelial scrape injury. Methods: Six weeks old New Zealand black mice were sacrificed and eyes enucleated at 4, 8, 16 ,24, and 48 hours after scrape removal of the epithelium. Eyes were fixed with 4% paraformaldehyde fixative for 4 hours and embedded in paraffin. Ten µm sections were then cut and mounted on slides. Antigen unmasking was performed by heat treatment in citrate buffer. The sections were permeabilized with 1% Triton X-100 for 15 minutes. Biotin-conjugated rat anti-mouse CD11b (1 µg/ml) (PharMingen) or rat anti-mouse T2008X (1µg/ml) (Research Diagnostics Inc.) was used to label monocytes. The Avidine-Biotin Complex method was used to visualize label (VectastainTM Elite Kit). Results: No monocytes were identified in unwounded control corneas. Within 8 hours of wounding small numbers of monocytes were present in the stroma. Large numbers of monocytes were present throughout the stroma at 24 and 48 hours after wounding. Monocytes were identified in the anterior stroma that was devoid of keratocytes from epithelial scrape-induced apoptosis. Conclusions: Corneal epithelial scrape injury triggers influx of large numbers of monocytes into the stroma. These inflammatory cells are likely attracted by chemokines produced by the remaining keratocytes and epithelial cells in response to injury.
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