A novel absorbable scaffold composed of chitosan and gelatin was fabricated by freezing and lyophilizing methods, resulting in an asymmetric structure. This bilaminar texture is suitable for preparing a bilayer skin substitute. The methods employed to confirm the applicability of this chitosan-gelatin scaffold as an ideal skin substitute were a water uptake ability test, in vitro fibroblast proliferation, and scaffold tests in which fibroblasts were co-cultured with keratinocytes. The chitosan-gelatin scaffolds were more wettable and adsorbed more water than did chitosan alone. In static cell culture the thinner scaffold is better than the thicker one, and because of diffusion limitations in the scaffold, culture time must be within 3 weeks before transplantation to living tissues. Keratinocytes were co-cultured with fibroblasts in chitosan-gelatin scaffolds to construct an artificial bilayer skin in vitro. The artificial skin obtained was flexible and had good mechanical properties. Moreover, there was no contraction observed in the in vitro cell culture tests. The data from this study suggest that chitosan-gelatin scaffolds are suitable for skin tissue engineering goals.
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