Improvement of Epidermal Barrier Properties in Cultured Skin Substitutes after Grafting onto Athymic Mice

Barrier function in cultured skin substitutes (CSS) prepared from human cell sources was measured by noninvasive (surface hydration, transepidermal water loss) and invasive methods (water permeation, niacinamide flux) before and after grafting onto athymic mice. In vitro measurements were made on days 7 and 14. Although three of the four measures of barrier function improved markedly from day 7 to 14, the values obtained were still far from those obtained with native human skin controls. Additional CSS were grafted onto athymic mice on day 14, and skin was harvested 2 and 6 weeks after grafting. Grafting brought about a substantial decrease in all measurements by 2 weeks and almost complete normalization of barrier function after 6 weeks. The most sensitive measure of this recovery was niacinamide permeability, which decreased from (280 ± 40) × 10–4 cm/h in vitro to (17 ± 30) × 10–4 cm/h 2 weeks after grafting and (5 ± 2) × 10–4 cm/h 6 weeks after grafting, versus control values of (2 ± 2) × 10–4 cm/h in human cadaver skin and (0.6 ± 0.4) × 10–4 cm/h in human epidermal membrane prepared from freshly excised breast skin. These results demonstrate the reformation of epidermal barrier function after transplantation and provide insights for the development of a functional epidermal barrier in CSS in vitro.

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