In Vitro Functional and Structural Evaluation of Low-Complexity Artificial Human Epidermis for 3D Tissue Engineering

In recent times, with the need for a reduction, refinement, and replacement of in vivo animal testing, there has been an increasing demand for the use of relevant in vitro human cell systems in drug development. There is also a great demand for the replacement of skin tissue in various wounds and burns. Furthermore, human skin cell-based in vitro systems can be used to investigate the side effects (toxicity and irritation) and tissue penetration of topical preparations. In this study, exploratory experiments were performed to produce artificial epidermis using two hydrogel scaffolds, alginate and GelMA C. The amount of keratinocytes added to the matrix (10-50-100 × 10⁶/mL) and the duration of tissue maturation (fresh, 1-3-4 weeks) were optimized in an extensive study. The behavior and structure of the two hydrogels were functionally and morphologically assessed. The permeability order for caffeine in the tested barriers was the following: alginate > GelMA C > cellulose acetate membrane > rat skin. It was concluded that GelMA C matrix provides a more favorable environment for cell survival and tissue differentiation (as demonstrated by histology and immunohistochemistry) than alginate. The 3-week incubation and 50 × 10⁶/mL cell number proved to be the most beneficial in the given system. This study provides data for the first time on the multifactorial optimization of two potential skin substitutes for tissue manufacturing. In order to use these results in tissue engineering, the fabricated artificial epidermis preparations must also be optimized for biocompatibility and from physical and mechanical point of views.