Development of a membrane and a bilayer of chitosan, gelatin, and polyhydroxybutyrate to be used as wound dressing for the regeneration of rat excisional wounds.

The skin is the largest organ in the human body that acts as a protective barrier from the outside environment. Certain dermatological pathologies or significant skin lesions can result in serious complications. Several studies have focused on the development of tissue-engineered skin substitutes. In this study, a new bilayer scaffold composed of a chitosan-gelatin membrane and a chitosan-polyhydroxybutyrate (PHB) porous matrix was synthesized and populated with human adipose-derived mesenchymal stem cells (hASCs) to be potentially used for wound dressing applications. By combining this membrane and porous matrix with the stem cells, we aimed to provide immunomodulation and differentiation capabilities for the wound environment, as well as mechanical strength and biocompatibility for the underlying tissue. The membrane was prepared from the mixture of chitosan and gelatin in a 2:1 ratio and the porous matrix was prepared from the mixture of chitosan and PHB, in equal proportions to form a final solution at 2.5% (m/v). Fourier transform infrared spectroscopy analysis showed the formation of blends, and micro-computed tomography, scanning electron microscopy and atomic force microscopy images demonstrated membrane roughness and matrix porosity. The MTT assay showed that the scaffolds were biocompatible with hASC. The membrane and the bilayer were used as dressing and support for cell migration in the dorsal excisional wound model in Wistar rats. Histological and gene transcriptional analyses showed that the animals that received the scaffolds regenerated the hair follicles in the deep dermis in the central region of the wound. Our results demonstrate the potential of these new biomaterials as dressings in wound healing studies, favoring tissue regeneration.