A bilayer strategy for wound healing: In vitro assessment of Alginate-PVA-BG hydrogel and AgNPs-loaded chitosan scaffolds

Objective(s): Developing multifunctional wound dressings requires integrating antimicrobial barriers with regenerative hydrogel matrices. This study engineered a bilayer scaffold comprising an upper electrospun chitosan (Cs)-polyethylene oxide (PEO) layer loaded with green-synthesized silver nanoparticles (AgNPs) and a lower lyophilized alginate (Alg)-polyvinyl alcohol (PVA) hydrogel layer enriched with bioactive glass (BG).Materials and Methods: AgNPs were synthesized using plant-derived proteolytic enzymes and characterized by DLS and UV-Vis spectroscopy. The electrospun upper layer was fabricated via electrospinning, while the lower hydrogel layer was prepared by freeze-drying. The Alg-PVA-BG hydrogel network was characterized for swelling behavior (gravimetrically at ۳۷ °C), degradation kinetics, porosity, and mechanical integrity. Biological evaluations included an MTT cytocompatibility assay using L۹۲۹ murine fibroblasts over ۱, ۳, and ۵ days, a wound closure assay using HaCaT human keratinocytes over ۴۸ hours, and antibacterial activity against Staphylococcus aureus and Escherichia coli using MIC۹۰ determination.Results: BG incorporation (۲۰% w/w) modulated the hydrogel structure, resulting in optimized porosity (۵۸-۶۷%), controlled swelling ratios (۱۴۷.۵-۱۸۹.۰%), and enhanced mechanical stability. The AgNP-loaded top layer (۳۲ µg/ml) exhibited potent broad-spectrum antibacterial activity with MIC۹۰ values of ۸.۲ µg/ml against S. aureus and ۱۲.۵ µg/ml against E. coli. In vitro biological evaluations demonstrated excellent cytocompatibility (cell viability >۸۵% for all groups) and significantly enhanced HaCaT cell migration (۴۲.۶۴±۵.۸۴%) for the Cs-PEO-AgNPs/Alg-PVA-BG group compared to the control group (P<۰.۰۵)..Conclusion: These results demonstrate the synergistic efficacy of combining an AgNP-mediated antimicrobial barrier with a bioactive alginate-based hydrogel matrix, offering a robust strategy for advanced wound-healing applications.

کلیدواژه ها:

Bioglass ، Hydrogel ، Nanoparticles ، Scaffold ، Wound healing