Burn injuries are common in both civilian and military settings and cause severe morbidity and mortality. The present study aimed to develop microporous bioactive scaffolds from the natural polysaccharide, pullulan, combined individually with Gelatin (Gel)/ Alginate (Alg)/ Chitosan (Ch)/ Carboxy-methyl cellulose (CMC), and crosslinked with glutaraldehyde to form a polymeric network. The objective was to evaluate and compare their efficacy in healing second degree partial-thickness burn wounds. The scaffolds were also impregnated with peppermint oil (PEO) to provide potential analgesic and cooling effects. Scaffolds were characterized using FTIR, TGA and SEM analysis to understand the intra-and intermolecular interactions and morphological characteristics. In vitro parameters like fluid uptake ability, biodegradation and hemolytic index were also assessed. Biocompatibility studies, including cell adhesion, migration and proliferation, were conducted using L929 mouse fibroblast cell lines. The in vivo efficacy was evaluated in Sprague Dawley rats inflicted with second degree burns to assess and compare wound retraction across different experimental groups. Histology, immunohistochemistry along with gram and picrosirius staining studies were also performed. The average pore size of all biopolymeric scaffold combinations ranged between 25.6-298μm. All four test combinations were non-hemolytic, with hemolytic indices between 0.3-2.9. Wound retraction studies in rats clearly indicated that among the four test combinations studied, Pu/Gel/PEO scaffold not only reduced local inflammation faster but also accelerated dermal regeneration in comparison to positive control as well as other test combinations (viz., Pu/Alg/PEO, Pu/Ch/PEO, and Pu/CMC/PEO). Immunohistochemistry results corroborated animal efficacy findings and demonstrated enhanced expression level of epidermal growth factor and fibroblast growth factor-2 in Pu/Gel/PEO scaffold treated group. Picrosirius staining also revealed enhanced collagen deposition in Pu/Gel/PEO scaffold treated group. The study findings suggest that biopolymeric scaffolds made from pullulan combined with gelatin and PEO (Pu/Gel/PEO) could serve as a promising substrate for skin regeneration template for second degree burns.
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