Perkins, M. A., D. A. Roberts, and R. Osborne. Development of In Vitro Methods for Use of Human Skin Cell Cultures for Skin and Eye Irritancy Assessments of Aqueous Incompatible Materials. J. Invest. Dermatol. 1992. 98(4): 638 (Abstract #517). [Reprinted by permission of Blackwell Science, Inc.].

In order to address the pressing need to reduce or replace the use of rabbits for prediction of human skin and eye irritancy potential, cultured human skin cells were evaluated. Currently available submerged cultures have limitations due to the aqueous buffered medium used for test material dilution. We have previously shown that these models are limited in their ability to predict irritancy potential for aqueous incompatible materials, product formulations, and acids/bases. To overcome these limitations, we developed a unique protocol utilizing Marrow-Tech Skin2 cultures for topical application of aqueous incompatible test materials as an alternative model for skin and eye irritancy testing. Methods were developed for topical application of and removal of neat test materials for acute damage evaluations. Materials tested in this model included a wide array of product formulation types, including liquids, gels, creams, foams, pastes, solids, powders and granulars. A battery of endpoints (MTT cell viability assay, lactate dehydrogenase enzyme release, and inflammatory mediator-PGE2 release,) was measured, representing final common pathways of skin or eye response to diverse test materials. Preliminary results indicate that, with this unique protocol, in vitro skin or eye irritancy assessments correlate well with the in vivo irritancy of these materials.