Background: Wound healing is a complex and overlapping process involving immune cells, cytokines, and growth factors. Objectives: This study aimed to design and evaluate a novel wound dressing based on postbiotic/chitosan in accelerating wound healing. Methods: Lactobacillus reuteri PTCC1655 was cultured, and the cell-free supernatant (postbiotic) was obtained by medium centrifugation. The films were prepared using the solvent casting method and evaluated in terms of water absorption index, water vapor transmission rate, and antimicrobial properties. Forty-five male Wistar rats were subjected to a full-thickness excisional wound to assess the wound healing potential. The rats were randomly divided into ctrl-, chitosan, and postbiotic groups. The time-course histological and gene expression analysis was performed to compare the dressing efficacy. Results: The films showed proper water absorption and water vapor transmission rate and inhibited the pathogens commonly associated with wound infection. The postbiotic film improved wound healing by modulating the inflammatory phase, increasing collagen and elastin deposition, and enhancing angiogenesis based on the histological results. The gene expression assay showed that the postbiotic film accelerated wound healing by improving the expression of inflammatory mediators (IL-6 and TNF-α) and anti-inflammatory mediators (TGF-β and VEGF). Conclusions: The cell-free supernatant/chitosan/polyethylene glycol (CFS/CS/PEG) biodegradable film could be introduced as a novel dressing for cutaneous wound healing. This transparent film enhances cutaneous wound healing by modulating infiltrated immunity cells and expressing inflammatory/anti-inflammatory cytokines.
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