The Center for Alternatives to Animal Testing is an academic center affiliated with the Division of Toxicological Sciences in the Department of Environmental Health Sciences of the Johns Hopkins University Bloomberg School of Public Health.

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Research Grants 2000-2001

Bovine Corneal Organ Culture: An Ex Vivo Model for Chemical Toxicity Tests

Fu-Shin Yu, PhD
The Schepens Eye Research Institute, Boston, Massachusetts

For five decades, the Draize test has remained the accepted method for evaluating the potential of test material to cause eye irritation or injury. Criticisms of this method center on the inhumaned treatment of animals and the irreproducibility of the subjective scoring procedure. There is a great demand for a mechanistic based in vitro testing system that will minimize the use of animals in chemical toxicity tests. Recently, we have developed an ex vivo model for chemical toxicity tests using a simple, long-term corneal organ culture method and tested several chemicals and consumer products. Our results showed that this system that closely resembles an in vivo testing, is an appropriate model for chemical safety tests. The corneas we use are prepared from the bovine eyes, economical and resourceful by-products of meat industry; no live animals are euthanized for testing. To perfect his system, our aims are:

To further elucidate how epithelial cells in organ culture respond to diverse classes of chemicals by assessing alteration of AP-1 and NF-B DNA-binding activity and normalize the data against known tonicities of test chemicals.
To investigate the changes in corneal function including impermeability and transparency and to correlate these with in vivo toxicity of known eye irritants.
To evaluate if recovery of epithelial functions after exposure of the corneal organ culture to a test chemical correlates with its ocular irritancy.

These ex vivo evaluations can be used for accurate prediction of irritation potential in vivo and offer a reliable alternative to the use of live animals.