Muller-Decker, K., G. Furstenberger, and F. Marks. Development of an In Vitro Alternative Assay to the Draize Skin Irritancy Test Using Human Keratinocyte-derived Proinflammatory Key Mediators and Cell Viability as Test Parameters. In Vitro Toxicol. 1992. 5(4): 191-209.

Tween 80 - 9005-65-6; acetone - 67-64-1; bradykinin - 58-82-2; benzalkonium chloride - 8001-54-5; sodium dodecyl sulfate - 151-21-3; ionophore A23187 - 52665-69-7

The aim of this study is to replace, at least partially, the animal Draize skin irritancy test by an in vitro cell culture test. Acute inflammation is known to be induced by the concerted release of various proinflammatory lipid metabolites and peptides. Keratinocytes representing the major cell type in skin are regarded to be the primary target for skin irritant action. The introduction of an in vitro alternative test critically depends on the selection of an appropriate battery of test parameters. Here, we propose to use three mechanistically independent in vitro parameters of irritancy which reflect major characteristics of acute inflammation i.e. the concentration of (I). key mediators of the eicosanoid cascade and (II). of interleukin-1a released upon challenging cells in vitro with test substances and (III). the toxic insult on these cells. The permanent human foreskin-derived keratinocyte cell line HPKII was used for the establishment of the routine test. In initial studies the test conditions were optimized with regard to the serum concentration and cell density. Upon challenge with exogenous irritant compounds HPKII are shown to release arachidonic acid and PGE2 and the proinflammatory cytokine interleukin-1a. Measuring proinflammatory mediator release and cytotoxicity in these cells may prove, therefore to be suitable as test system for the prediction of the irritant potential of chemicals.