Abstract
Antimicrobial resistance (AMR) and biofilm formation significantly hinder chronic wound management, necessitating safer and more effective therapeutic options. This study evaluates the antimicrobial, antibiofilm, cytocompatibility and anti-inflammatory properties of a novel antimicrobial hydrogel Formulation (#1) compared with commercially available wound hydrogel and cream Formulations (#2-5). Antimicrobial activity was assessed using minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) assays against Staphylococcus aureus, Pseudomonas aeruginosa, Candida albicans, and mixed-species cultures. Biofilm-related efficacy was evaluated via crystal violet (CV) staining and minimum biofilm inhibitory concentration (MBIC) assays. Cytotoxicity was examined using ISO-compliant MTT and confluency assays on L929 fibroblasts. In vivo anti-inflammatory effects were assessed using intra-cutaneous injection in New Zealand White rabbits with histological evaluation per ISO 10993-23. Formulation 1 showed the lowest MIC and MBC values across all pathogens, including polymicrobial cultures, indicating strong broad-spectrum efficacy. In biofilm assays, it reduced biofilm biomass by 50%-60% within 10 min and prevented new formation at lower MBIC values than Formulation 2, especially in mixed-species models. Cytotoxicity testing confirmed Formulation 1 maintained ≥ 81% cell viability across all concentrations, outperforming other products and meeting ISO and USP safety thresholds. In vivo, both Formulation 1 and 2 induced minimal inflammation, with Formulation 1 showing slightly milder tissue responses. Formulation 1 demonstrated strong antimicrobial efficacy, reliable biofilm control, and favourable cytocompatibility compared with the comparator formulations tested in this study. These findings support further evaluation of this formulation for chronic wounds complicated by biofilms and antimicrobial resistance.