An in Vitro Method for Assessing Corneal Opacification Potential Using a Rabbit Corneal Cell Line

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Abstract
We report an in vitro method for identifying materials with the potential to cause corneal opacity. Monolayers obtained from an established line of rabbit corneal cell epithelium were exposed to test materials and the increase in light absorbance at 360 nm was assessed after 10 min. Responses obtained proved to be concentration-dependent and the curves generated were of sufficiently different character to allow differentiation among the test materials. Because trichloroacetic acid (2%) caused a linear response over the entire range of initial monolayer densities used and provided the widest degree of response of any of the materials tested, it was chosen as a reference compound. The effects of all other test materials were expressed in relation to that observed for 2% trichloroacetic acid to provide a basis for comparative effects and to control for the effects of intertrial variation in monolayer density. The increased absorbance caused by 20% acetaldehyde and 2% silver nitrate was 30% and 60%, respectively, of that produced by 2% trichloroacetic acid. Formaldehyde in concentrations up to 20% produced no acute change in monolayer absorbance in this test system. When tricholoracetic acid was used as the test material (when compared to itself), the effect of a 2% concentration was similar to that predicted. Results obtained were related to published reports of the in vivo effects of the test materials. However, no in vivo or in vitro studies reported have specifically ranked the effects of the test materials used herein.
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