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ToxSci Advance Access originally published online on February 18, 2003
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Toxicological Sciences 72, 122-129 (2003)
Copyright © 2003 by the Society of Toxicology

IN VITRO TOXICOLOGY AND ALTERNATIVE TESTING

Resazurin Metabolism Assay Is a New Sensitive Alternative Test in Isolated Pig Cornea

Sébastien Perrot*,{ddagger}, Hélène Dutertre-Catella*,{dagger}, Chantal Martin*, Patrice Rat*,{ddagger} and Jean-Michel Warnet*,{ddagger},1

* Laboratoire de Toxicologie, Faculté des Sciences Pharmaceutiques et Biologiques, Université Paris V, 75270 Paris cedex 06, France; {dagger} Laboratoire de Toxicologie, Faculté des Sciences Pharmaceutiques, Université de Tours, 37200 Tours, France; and {ddagger} Unité de Pharmaco-Toxicologie Cellulaire, Centre Hospitalier National d’Ophtalmologie des XV-XX, 75571 Paris cedex 12, France

The main object of our study was to investigate whether the resazurin metabolism assay is a sensitive surfactant and alcohol toxicity test in isolated pig cornea and to compare this recently developed fluorometric assay with the data collected in the eye irritation reference chemical data bank. Resazurin is a substrate that changes color in response to metabolic activity. Isolated pig corneas were immersed for 10 min in surfactants and alcohol irritant solutions. After incubation, resorufin fluorescence was read and corneal viability was assessed. This corneal viability was compared with the maximal modified average score published in the report of ECETOC. This assay highlighted different concentration-dependent irritation potentials of the three surfactants tested, and the same results were obtained with corneas treated with the alcohols. We observed that the degree of surfactant- and alcohol-induced decrease in corneal viability, using the resazurin reduction test, was correlated with the in vivo irritancy measurements as determined by the Draize test and scored with the Modified Maximum Average Score (MMAS). This assay allowed us to classify the ocular irritancy of the tested surfactants and alcohols in the same ranking order as the Draize classification. Corneal viability measurement can be used as a potential alternative for the toxicological assessment of surfactants and alcohols. The nontoxic, nonradioactive resazurin metabolism assay allows rapid assessment of many samples with simple equipment and at reduced cost for continuous monitoring of corneal viability. This assay seems to be suitable as a toxicological screening test for eye irritation determination.

Key Words: isolated corneal model; toxicological assessment; resazurin; surfactants; alcohols.


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