Fabrication of Nanofibrous/Xerogel Layer-by-Layer Biocomposite Scaffolds for Skin Tissue Regeneration: In Vitro Study

[Image: see text] Skin burn wounds are a crucial issue that could reduce life quality. Although numerous effective skin products have invaded the biomedical market, most of them still demonstrate some limitations regarding their porosity, swelling and degradation behaviors, antibacterial properties, and cytotoxicity. Thus, the aim of this study is to fabricate novel trilayered asymmetric porous scaffolds that can mimic the natural skin layers. In particular, the fabricated scaffold constitutes an upper electrospun chitosan–poly(vinyl alcohol) layer and a lower xerogel layer, which is made of effective skin extracellular matrix components. Both layers are fixed together using fibrin glue as a middle layer. The results of this study revealed promising scaffold swelling capability suitable for absorbing wound exudates, followed by a constant degradable weight over time, which is appropriate for a burn wound environment. Scanning electron microscopy images revealed an average pore diameter in the range of 138.39–170.18 nm for the cross-linked electrospun mats and an average pore size of 2.29–30.62 μm for the fabricated xerogel layers. This further provided an optimum environment for fibroblast migration and proliferation. The electrospun nanofibrous layer was examined for its antibacterial properties and showed expressive complete bacterial inhibition against Gram-positive (Staphylococcus aureus) and Gram-negative (Escherichia coli) bacterial strains (log reduction = 3 and 2.70, respectively). Next, mouse embryonic fibroblast cytotoxicity and migration rate were investigated against the developed asymmetrical composite to assess its biocompatibility. Tissue culture experiments demonstrated significant cell proliferation and migration in the presence of the constructed scaffold (P < 0.0001). A complete wound closure was observed in vitro in the presence of the three scaffold asymmetrical layers against the mouse embryonic fibroblast. The results of this study proved superior biological characteristics of the innovative asymmetrical composite that could further replace the burned or damaged skin layers with promising potential for clinical applications.