S Abadie1, P Bedos1, J Rouquette2. 1. Syntivia, Centre Pierre Potier, 1 place Pierre Potier, Entrée B, BP 50624, 31106, Toulouse, France. 2. ITAV, Centre Pierre Potier, Université de Toulouse, CNRS, UPS, 1 place Pierre Potier, Entrée B, BP 50624, 31106, Toulouse, France.
Abstract
OBJECTIVE: The main function of skin is to protect the body from external aggressions. Over time, normal skin ageing is accelerated by external stresses such as smoking, pollution, chemical products and radiation. UV light, in particular UVA, causes DNA damage, apoptosis and morphological modifications, which are responsible for both premature ageing and cancer. The aim of this study was to establish a discriminatory and sufficiently reproducible cutaneous model for evaluating UVA damage, to enable testing for effectiveness of potentially protective compounds. METHODS: The cutaneous model is based on Human skin explants irradiated with UVA. Deleterious effects on epidermis were observed and quantified by haematoxylin-eosin staining and by immunofluorescence of ɣ-H2Ax, cytokeratin 5, involucrin and loricrin protein. Dermis deterioration was evaluated by transmission electronic microscopy and zymography in situ. RESULTS: We were able to observe and quantify deleterious effects associated with UVA irradiation: epidermal and dermal disruption, appearance of Sunburn cells, increased DNA damage and induced apoptosis. The use of this model in the evaluation of protective compounds was first confirmed using sunscreens, then further validated with a panel of active ingredients which showed beneficial effects on epidermis morphology and DNA integrity after UVA exposure. CONCLUSION: We have developed a model and a standardized protocol, based on the use of human skin explants, which allows us to explore the protective effect of active ingredients to environmental stresses such as UVA.
OBJECTIVE: The main function of skin is to protect the body from external aggressions. Over time, normal skin ageing is accelerated by external stresses such as smoking, pollution, chemical products and radiation. UV light, in particular UVA, causes DNA damage, apoptosis and morphological modifications, which are responsible for both premature ageing and cancer. The aim of this study was to establish a discriminatory and sufficiently reproducible cutaneous model for evaluating UVA damage, to enable testing for effectiveness of potentially protective compounds. METHODS: The cutaneous model is based on Human skin explants irradiated with UVA. Deleterious effects on epidermis were observed and quantified by haematoxylin-eosin staining and by immunofluorescence of ɣ-H2Ax, cytokeratin 5, involucrin and loricrin protein. Dermis deterioration was evaluated by transmission electronic microscopy and zymography in situ. RESULTS: We were able to observe and quantify deleterious effects associated with UVA irradiation: epidermal and dermal disruption, appearance of Sunburn cells, increased DNA damage and induced apoptosis. The use of this model in the evaluation of protective compounds was first confirmed using sunscreens, then further validated with a panel of active ingredients which showed beneficial effects on epidermis morphology and DNA integrity after UVA exposure. CONCLUSION: We have developed a model and a standardized protocol, based on the use of human skin explants, which allows us to explore the protective effect of active ingredients to environmental stresses such as UVA.