A microelectric cell sensing technique for in vitro assessment of ocular irritation

Abstract

The animal-based Draize test remains the gold standard for assessment of ocular irritation. However, subjective scoring methods, species differences, and animal welfare concerns have spurred development of alternative test methods. In this study, a novel in vitro method for assessing ocular irritancy was developed using a microelectric cell sensing technology, real-time cell analysis (RTCA). The cytotoxicity of sixteen compounds was assessed in two cell lines: ARPE-19 (human retina) and SIRC (rabbit cornea). In vitro inhibitory (IC50 and AUC50) values were determined at 6, 12, 24, 48, 72, and 96 h exposure, with a subset of values confirmed with MTT testing. The values displayed comparable predictivity of in vivo ocular irritation on the basis of a linear regression between the calculated values and each compounds' corresponding Draize-determined modified maximum average score (MMAS), but the ARPE-19 derived values were more strongly correlated than those from SIRC cells. Hence, IC50 values derived from ARPE-19 cells were used to predict the UN GHS/EU CLP classification of each test compound. The method was determined to have sensitivity of 90%, specificity of 50%, and overall concordance of 75%. Thus, RTCA testing may be best incorporated into a top-down tiered testing strategy for identification of ocular irritants in vitro.