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J. Echegaray, M. Senchyna, D. L. Meadows, H. A. Ketelson, V. L. Perez; Real Time in vivo Model to Quantify the Time Course and Magnitude of Inflammatory Cell Recruitment to the Cornea Following Topical Exposure to Ophthalmic Preservatives. Invest. Ophthalmol. Vis. Sci. 2010;51(13):1571.
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To utilize in vivo fluorescence stereomicroscopy to track the recruitment of fluorescently labeled inflammatory cells of Enhanced Green Fluorescent Protein (EGFP) chimeric mice during topical ocular treatment with preservatives such as benzalkonium chloride (BAC).
EGFP mice were generated as previously described. One eye of each EGFP mouse was dosed with 0.5 % BAC, where as the contra lateral eye was dosed with saline (control eye). Two different topical application strategies were tested: one group of mice (n=3) was dosed QID daily for 15 days and a second group of mice (n=3) was dosed with a single drop at days 0, 3, 5, 7, 10 and 15 while under anesthesia. Recruitment of inflammatory cells was monitored by in vivo fluorescent microscopy and digital photography at days 1, 3, 5, 7, 10, 15 and 25. Corneal haze and scaring was qualitatively evaluated at similar time points.
Recruitment of EGFP labeled inflammatory cells was detected in BAC treated corneas as early as day 3 and continued to increase in magnitude over the course of the dosing period in both topical application models. Animals dosed under anesthesia displayed a higher number of EGFP cells in the cornea at earlier time points. Significant infiltration was not detected in contra lateral control eyes of EGFP mice in either dosing model at any timepoint. In both groups, the magnitude of EGFP cell corneal infiltration appeared to correlate with the degree of corneal haze and scarring.
We have demonstrated that corneal infiltration of immune cells occurs in response to exposure to preservatives such as BAC. Ongoing efforts are aimed at identifying individual populations of cells involved in the recruitment process. Clinically, the degree of corneal infiltration appears related to signs of ocular surface damage. This model will provide significant insight into the inflammatory processes associated with ocular surface toxicity and enable the development of alternative preservatives for use in ophthalmic products.
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