Assessment of the skin barrier function in the reconstructed human epidermis using a multimodal approach at molecular, tissue and functional levels
Résumé
Reconstructed human epidermis models are used as epidermis alternatives in skin researches. It is necessary to provide molecular and functional characterization in order to assess these models. Our aim is to make a link between the barrier function and the structure and the composition of the Stratum Corneum using several complementary techniques. Three studies were used on Reconstructed Human Epidermis during the keratinocyte differentiation process: i- caffeine percutaneous penetration kinetic. ii- epidermis thickness measurement, Stratum Corneum formation and lipids organization by Raman microspectroscopy. iii- lipid composition evolution by liquid chromatography coupled to high resolution mass spectrometry. Results demonstrated that the caffeine penetration decreased along the differentiation process. Raman in depth images demonstrated an increase in Stratum Corneum and RHE thickness accompanied with an evolution of lipid organization. Lipid analysis showed an increase of ceramide amount and an inverse relationship between ceramide and their precursor’s levels during the differentiation process. A different behavior between several ceramides subclasses is highlighted and relied to corresponding differentiation stage. The generation of the most important ceramides for the barrier function is closely followed. A period shift between lipids generation and their organization was found. Our analytical data allowed identifying 3 groups of maturation days: before day 15, between day 15 and 19, and after day 19. The chemical and physiological states of the barrier function for each group are described thanks to the multimodal approach.
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Assessment of akin barriere- analyst - 3ème soumission RK RJ JB AT-2-version Hal.pdf (1.14 Mo)
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