Abstract
An in vitro human reconstructed epidermis model (SkinEthic) used for screening acute and chronic skin irritation potential was validated against in vivo data from skin tolerability studies. The irritation potential of sodium lauryl sulfate (SLS), calcipotriol and trans-retinoic acid was investigated. The in vitro epidermis-like model consists of cultures of keratinocytes from human foreskin on a polycarbonate filter. The modulation of cell viability, the release and gene expression of proinflammatory cytokines, interleukins 1α and 8, and morphological changes were evaluated during 3 days as endpoints representative for an inflammatory reaction. The cumulative irritation potential of the topical products was evaluated in a human clinical study by visual scoring and biophysical measurement of inflammatory skin reaction after repeated 24 h applications over 3 weeks under Finn chamber patches. All topical products that were nonirritating in the human study were noncytotoxic and did not induce cytokine expression in the in vitro acute model (day 1 exposure). All irritating controls exhibited specific cell viability and cytokine patterns, which were predictive of the in vivo human data. The ranking of mild to moderate skin irritation potential was based on the lack of cytotoxicity and the presence of cytokine patterns including gene expression specific for each irritant, using the chronic in vitro model (up to 3 days exposure).
The human reconstructed epidermis model SkinEthic was shown to be a reliable preclinical tool predicting the irritation potential of topical products. Moreover, it is a useful model in a two-step tiered strategy for screening acute and chronic irritation potential for the selection of vehicles for new topical drugs.
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References
Basketter DA, Griffiths HA, Wang XM, Wilhem KP, Mac Fadden J. Individual, ethnic and seasonal variability in irritant susceptibility of skin: the implications for a predictive human patch test. Contact Dermatitis. 1996;35:208–13.
Barker JNWN, Jones ML, Mitra RS, et al. Modulation of keratinocyte-derived interleukin-8 which is chemotactic for neutrophils and T-lymphocytes. Am J Pathol. 1991;139:869–76.
Bell E, Parenteau N, Gay R, et al. The living skin equivalent: its manufacture, its organotypic properties and its responses to irritants. Toxicol in Vitro. 1991;5:591–6.
Boelsma E, Tanojo H, Boddé HE, Ponec M. Assessment of the potential irritancy of oleic acid on human skin: evaluation in vitro and in vivo. Toxicol in Vitro. 1996;10:729–42.
Boelsma E, Tanojo H, Boddé HE, Ponec M. An in vivo-in vitro study of the use of a human skin equivalent for irritancy screening of fatty acids. Toxicol in Vitro. 1997;11:365–76.
Bos JD, Kapsenberg ML. The skin immune system: progress in cutaneous biology. Immunol Today. 1993;14:75–8.
Botham PA, Earl LK, Fentem JH, Roguet R, van de Sandt JJM. Alternative methods for skin irritation testing: the current status. ATLA. 1998:195–211.
Boyce S, Michel S, Reichert U, Shroot B, Schmidt R. Reconstructed skin from cultured human keratinocytes and fibroblasts on a collagen glycosamine biopolymer substrate. Skin Pharmacol. 1990;3:136–43.
Braa SS, Triglia D. Predicting ocular irritation using 3-dimensional human fibroblast cultures. Cosmetics Toiletries 1991;106:55–60.
Cooper II JA, Saracci R, Cole P. Describing the validity of carcinogen screening tests. Br J Cancer. 1979;39:87–9.
Corsini E, Bruccoleri A, Marinovich M, Galli CL. Endogenous interleukin-1 alpha associated with skin irritation induced by tributylin. Toxicol Appl Pharmacol. 1996;138(2):268–74.
Draize JH, Woodard G, Avery HO. Methods for the study of irritation and toxicity of substances applied topically to the skin and mucous membranes. J Pharmacol Exp Ther. 1944;82:377–90.
Dinarello CA. Interleukin 1 and interleukin 1 antagonism. Blood. 1991;77:1627–52.
Doucet O, Robert C, Zastrow L. Use of a serum free reconstituted epidermis as a skin pharmacological model. Toxicol in Vitro. 1996;10:305–13.
Emmett EA. Toxic responses of the skin. In: Casarett and Doull's toxicology. New York: Macmillan; 1986:412–31.
Fullerton A, Benfeldt E, Petersen JR, Jensen SB and Serup J. The calcipotriol dose-irritation relationship: 48 hour occlusive testing in healthy volunteers using Finn chambers. Br J Dermatol. 1998;138:259–65.
Griffiths CEM, Barker JNWN, Kunkel S, Nickoloff BJ. Modulation of leucocyte adhesion molecule, a T cell chemotaxin (IL-8) and a regulatory cytokine (TNF-α) in allergic contact dermatitis (rhus dermatitis). Br J Dermatol. 1991;124:519–26.
Kligman AM. The identification of contact allergens by human assay. J Invest Dermatol. 1966;47:393–409.
Kligman AM, Grove GL, Hirose R, Leyden JL. Topical tretinoid for photoaged skin. J Am Acad Dermatol. 1986;15: 836–59.
Kubilus J, Cannon CL, Neal PJ, Ricker HA, Klausner M. Cytokine response of the Epiderm skin model to topically applied irritants and toxicants. In Vitro Toxicol. 1996;9:157–66.
Kupper TS, Groves RW. The interleukin-1 axis and cutaneous inflammation. J Invest Dermatol. 1995;105:62S-6S.
Naughton GK, Jacob L, Naughton BA. A physiological skin model for in vitro toxicity studies. In: Goldberg AM, ed. Alternative methods in toxicology. New York: Mary Ann Liebert. 1989;7:183–9.
Queille-Roussel C, Duteil L, Padilla JM, Poncet M, Czernielewski J. Objective assessment of topical anti-inflammatory drug activity on experimentally induced nickel contact dermatitis: comparison between visual scoring, colorimetry, laser Doppler velocimetry and transepidermal water loss. Skin Pharmacol. 1990;3:248–55.
Régnier M, Darmon M. Human epidermis reconstructed in vitro: a model to study keratinocyte differentiation and its modulation by retinoic acid. In Vitro Cell Dev Biol. 1989;25:1000–8.
Roguet R, Cohen C, Dossou KG, Rougier A. Episkin, a reconstituted human epidermis for assessing in vitro the irritancy of topically applied compounds. Toxicol in Vitro 1994;8:283–91.
Roguet R, Cohen C, Robles C et al. An interlaboratory study of the reproducibility and relevance of Episkin, a reconstructed human epidermis, in the assessments of cosmetics irritancy. Toxicol in Vitro 1998;12:295–304.
Rosdy M, Claus L-C. Terminal differentiation of human keratinocytes grown in chemically defined medium on inert filter substrates at the air-liquid interface. J Invest Dermatol. 1990;95:409–14.
Tinois E, Tiollier J, Gaucherand M, Dumas H, Tardy M, Thivolet J. In vitro and post-transplantation differentiation of human keratinocytes grown on human type IV collagen film of a bilayered dermal substitute. Exp Cell Res. 1991;193:310–19.
Van der Valk PGM, Maibach HI. The irritant contact dermatitis syndrome. Boca Raton, FL: CRC Press, 1995.
York M, Griffiths MA, Whittle E, Basketter DA. Evaluation of a human patch test for the identification and classification of skin irritation potential. Contact Dermatitis. 1996;34:204–12.
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de Brugerolle de Fraissinette, A., Picarles, V., Chibout, S. et al. Predictivity of an in vitro model for acute and chronic skin irritation (SkinEthic) applied to the testing of topical vehicles. Cell Biol Toxicol 15, 121–135 (1999). https://doi.org/10.1023/A:1007577515215
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DOI: https://doi.org/10.1023/A:1007577515215