Development of a multiparametric in vitro model of skin sensitization

• Published in: Journal of applied toxicology 2015-01, Vol.35 (1), p.48-58
• Main Authors: Guyard-Nicodème, Muriel, Gerault, Eloise, Platteel, Marion, Peschard, Olivier, Veron, Wilfried, Mondon, Philippe, Pascal, Svinareff, Feuilloley, Marc G. J.
• Format: Article
• Language: English
• Subjects: Allergens - chemistry; Allergens - toxicity; Animal Testing Alternatives; Bacteriology; Biocompatibility; Biomedical materials; Cell Behavior; Cell Survival - drug effects; Cells, Cultured; Cellular Biology; Correlation; cosmetic compounds; Cosmetics; Cosmetics - chemistry; Cosmetics - toxicity; Cytokines; Dermatitis, Allergic Contact - etiology; Dermatitis, Allergic Contact - immunology; Diffusion Chambers, Culture; Haptens - metabolism; Humans; in vitro models; In Vitro Techniques; In vitro testing; interleukin 18; Interleukin-18 - biosynthesis; Interleukin-18 - immunology; Irritants - chemistry; Irritants - toxicity; Keratinocytes - drug effects; Keratinocytes - immunology; Keratinocytes - pathology; Life Sciences; Mathematical models; Microbiology and Parasitology; Models, Statistical; MTT Test; Multivariate Analysis; Skin - drug effects; Skin - immunology; Skin - pathology; Skin Absorption - drug effects; skin permeation; skin sensitization; Toxicity; Toxicity Tests, Acute - methods; Toxicology; xenobiotic haptenation
• ISSN: 0260-437X; 1099-1263
• DOI: 10.1002/jat.2986

ABSTRACT Most animal experiments on cosmetics safety are prohibited and since March 2013, this obligation includes sensitization tests. However, until now there has been no validated alternative in vitro method. In this work, 400 compounds used in the cosmetic industry were selected to cover the greatest diversity of structures, biological activities and sensitizing potential. These molecules were submitted to a series of tests aimed at reproducing essential steps in sensitization and to distinguish between sensitization and irritations, i.e., transcutaneous permeation (factor A), haptenation (factor B), sensitization cytokines production (factor C) and acute toxicity (factor D). The transcutaneous diffusion was measured on human skin explants using Franz cells. Haptenation was tested in solution on human serum albumin. Sensitization cytokine production was investigated by measurement of interleukin‐18 release by keratinocytes. Acute toxicity was determined using an 3‐(4,5‐Dimethylthiazol‐2‐yl)‐2,5‐diphenyltetrazolium bromide75 cell viability test. As only sufficiently stable, soluble and detectable compounds are usable, 33, 72, 68 and 68 molecules were finally tested on factors A, B, C and D, respectively, and 32 were completely screened by the four factors. The individual correlation of the four factors with the reference in vivo tests was limited but the combination of these factors led to a correlation between in vivo and in vitro assays of 81.2% and the safety of the test (risk of false negative) reached 96.8%. The techniques employed are simple and inexpensive and this model of four tests appears as a promising technique to evaluate in vitro the skin sensitization potential of unknown molecules. Copyright © 2014 John Wiley & Sons, Ltd. A new in vitro model of skin sensitizing potential based on the measure of 4 parameters, i.e. trans‐cutaneous permeation (Factor A), haptenation (Factor B), sensitization cytokines production (Factor C) and acute toxicity (Factor D) was described. A total of 32 compounds regularly employed in cosmetics were tested in this model. The correlation with in vivo assays reached 81.2% and the safety of the test (risk of false negative) reached 96.8 %.