title: Effect of skin barrier competence on SLS and water-induced IL-1alpha expression.
abstract: For screening of a potential irritant it is essential that an early marker for irritation should be chosen which could be detected before the physiological signs of irritation occur. Interleukin 1 alpha (IL-1alpha) is widely accepted as such a marker in both in vivo and in vitro test systems. In this study, we have determined the mRNA levels of IL-1alpha in the epidermis after topical application of sodium dodecyl sulphate (SLS) in both a commercially available epidermal kit (EpiDerm) and in excised skin. Furthermore, we have determined the effect of water, the vehicle for SLS, on IL-1alpha mRNA levels. Topical application of water to excised skin increases IL-1alpha mRNA levels sixfold in the epidermis whereas topical application of water to EpiDerm cultures did not alter IL-1alpha mRNA levels. This is explained by the finding that EpiDerm cultures have a sub-optimal barrier function when compared with excised skin - topical application of SLS was clearly toxic at much lower concentrations in EpiDerm cultures (0.2% SLS) than in excised skin (5% SLS). Also caffeine penetration was 10-fold higher through EpiDerm cultures than through the excised skin. Therefore, incubation of control EpiDerm cultures at 100% humidity effectively mimics topical exposure to water. An additional increase in IL-1alpha mRNA levels observed between topical application of water and SLS is similar (about threefold) in both experimental systems. In conclusion, in vitro reconstructed epidermis models, such as EpiDerm, can be used as a predictive model for irritancy screening. However, great care should be taken when interpreting the results due to the fact that EpiDerm cultures do not have a competent barrier function and therefore lower irritant concentrations are required than in in vivo or ex vivo studies in order to induce cytotoxic effects. Furthermore, the irritant effects of the vehicle should not be neglected. Our results show clearly that the topical application of water to excised skin results in increased levels of IL-1alpha mRNA in the epidermis. This is a cytokine that is widely used as an early marker for skin irritation.
authors.affiliation: Department of Dermatology, Leiden University Medical Centre, The Netherlands. S.Gibbs@VUMC.nl
authors.forename: Susan
authors.initials: S
authors.lastname: Gibbs
authors.forename: Helene
authors.initials: H
authors.lastname: Vietsch
authors.forename: Ursi
authors.initials: U
authors.lastname: Meier
authors.forename: Maria
authors.initials: M
authors.lastname: Ponec
mesh_terms: D001483:Base Sequence; D002463:Cell Membrane Permeability / Q000276:immunology*; D002470:Cell Survival / Q000187:drug effects; D017931:DNA Primers; D004817:Epidermis / Q000187:drug effects / Q000276:immunology; D005786:Gene Expression Regulation / Q000187:drug effects / Q000276:immunology*; D006801:Humans; D007375:Interleukin-1 / Q000235:genetics*; D012339:RNA, Ribosomal, 28S / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012879:Skin Physiological Phenomena; D012967:Sodium Dodecyl Sulfate / Q000494:pharmacology*; D014867:Water
chemical_list: D017931:DNA Primers; D007375:Interleukin-1; D012339:RNA, Ribosomal, 28S; D014867:Water; D012967:Sodium Dodecyl Sulfate