Corneal epithelial wound closure in tissue culture: an in vitro model of ocular irritancy

The influence of 13 test agents on the ability of cultured rabbit corneal epithelial cells to migrate and re-cover a wound has been utilized to evaluate an in vitro model of ocular irritancy. Cells were grown under standard conditions and monitored for adequate cell density. Seven days after subculture, replicate wounds were produced and cultures were exposed to varying concentrations of a test agent in culture medium for 24 hr. Following exposure, cultures were fixed and stained to reveal remaining wound areas which were quantitated by computerized planimetry and compared to evaluate the deleterious effects of the test agents. The test ranks irritants in an order similar to that described in the literature for both in vivo and in vitro tests. This tissue culture model is conceptually simple, quantitative, and an alternative to the corneal component of whole animal testing for ocular irritancy.