Predicting ocular irritancy and recovery from injury using Madin-Darby canine kidney cells

Two promising cell culture assays, using Madin-Darby canine kidney cells, for predicting eye irritancy, the fluorescein leakage assay and the neutral red release assay, have been adapted to try and assess the ability of damaged cells to recover from chemical-induced injury. The fluorescein leakage and neutral red release protocols are similar but measure injurious effects on different parts of the cells, namely the tight junctions and the cell membrane, respectively. Both endpoints have previously given equivalent rankings of chemicals in order of their eye irritancy potential. Sixteen compounds of varying irritancy potential and chemical nature were tested using the two assays. In both assays, little or no cell recovery was measured 72 hr after a mildly injurious exposure, although using the fluorescein leakage assay, five test agents displayed substantial recovery and two displayed significant deterioration of the cell layer after removal of the test material. Comparing these in vitro results with in vivo data suggests that the fluorescein leakage assay, in its current format, does not predict the likely recovery rate of ocular tissue after chemical damage.