title: Keratinocyte-derived proinflammatory key mediators and cell viability as in vitro parameters of irritancy: a possible alternative to the Draize skin irritation test.
abstract: This study is aimed at the development of a cell culture assay which may supplement or replace the animal Draize skin irritancy test. Using human keratinocytes, the measurement of proinflammatory eicosanoid and interleukin-1 alpha release and of the impairment of cell viability have provided a suitable in vitro/in vivo correlation for at least three surfactants. The in vitro study has been extended using structurally unrelated, pharmacologically relevant compounds including ethanol, glycerol, cyclohexanol, acetone, benzoic acid, phenol, acrylamide, triethanolamine, Tween 80, sodium dodecyl sulfate, benzalkonium chloride, NiSO4, SnCl2, and ZnCl2. Time- and dose-response studies were used to establish half-maximal stimulatory (SC50) and inhibitory (IC50) as well as 10-fold stimulatory (ED10) concentrations for arachidonic acid release, cytotoxicity, and IL-1 alpha release, respectively. Based upon these values a similar ranking has been obtained for the mildly acting acetone, ethanol, glycerol, and Tween 80 and for the severely acting A23187, benzalkonium chloride, and sodium dodecyl sulfate. With respect to all other test compounds, substantial variation occurred indicating that all three test parameters provide a more complete characterization of the test compound's potency than a single endpoint. These data are ready to be validated by a controlled clinical study aimed at a qualitative and quantitative evaluation of the symptoms of skin inflammation in volunteers.
authors.affiliation: German Cancer Research Center, Research Program Tumor Cell Regulation, Heidelberg.
authors.forename: K
authors.initials: K
authors.lastname: Müller-Decker
authors.forename: G
authors.initials: G
authors.lastname: Fürstenberger
authors.forename: F
authors.initials: F
authors.lastname: Marks
mesh_terms: D000826:Animal Testing Alternatives; D000818:Animals; D016718:Arachidonic Acid / Q000378:metabolism*; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D002851:Chromatography, High Pressure Liquid; D004305:Dose-Response Relationship, Drug; D006801:Humans; D007375:Interleukin-1 / Q000378:metabolism*; D007509:Irritants / Q000633:toxicity*; D015603:Keratinocytes / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D050356:Lipid Metabolism; D008928:Mitochondria / Q000201:enzymology; D013385:Succinate Dehydrogenase / Q000378:metabolism
chemical_list: D007375:Interleukin-1; D007509:Irritants; D016718:Arachidonic Acid; D013385:Succinate Dehydrogenase