Abstract
Healthy skin moLEdels produced by tissue-engineering often present a suboptimal skin barrier function as compared with normal human skin. Moreover, skin substitutes reconstructed according to the self-assembly method were found to be deficient in polyunsaturated fatty acids (PUFAs). Therefore, in this study, we investigated the effects of a supplementation of the culture media with docosahexaenoic acid (DHA) on the barrier function of skin substitutes. To this end, 10 μM DHA-supplemented skin substitutes were produced (n = 3), analyzed, and compared with controls (substitutes without supplementation). A Franz cell diffusion system, followed by ultra-performance liquid chromatography, was used to perform a skin permeability to testosterone assay. We then used gas chromatography to quantify the PUFAs found in the epidermal phospholipid fraction of the skin substitutes, which showed successful DHA incorporation. The permeability to testosterone was decreased following DHA supplementation and the lipid profile was improved. Differences in the expression of the tight junction (TJ) proteins claudin-1, claudin-4, occludin, and TJ protein-1 were observed, principally a significant increase in claudin-1 expression, which was furthermore confirmed by Western blot analyses. In conclusion, these results confirm that the DHA supplementation of cell culture media modulates different aspects of skin barrier function in vitro and reflects the importance of n-3 PUFAs regarding the lipid metabolism in keratinocytes.
Highlights
The skin forms the interface between the body and the external environment
tight junction (TJ) control the flow of molecules through the intercellular space between the cells of the epidermis, so that compounds must enter the cells by diffusion or active transport in order to pass through the tissue [6]
In a previous study by our team, we have shown that the supplementation of the culture media with alpha-linolenic acid (ALA) decreased the percutaneous absorption of testosterone through the skin substitutes [18]
Summary
The skin forms the interface between the body and the external environment. Its main function is to protect the organism against extraneous aggressions by forming a vital and waterproof barrier, preventing the onset of an immune response [1]. The barrier function is mainly provided by the outermost layer of the skin, namely the epidermis, and more precisely by the stratum corneum (SC) [2]. The corneocytes are embedded in a lipid matrix, which ensures the maintenance of an optimal paracellular skin barrier function. The three main lipids that dominate the SC lipid matrix are cholesterol, free fatty acids, and ceramides, which adopt a highly ordered structure. They are arranged perpendicularly to the surface of the corneocytes, parallel to each other, on the extracellular side of the cornified envelope [4].
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