Fabrication, characterization, and biocompatibility studies of formononetin-loaded chitosan-oxidized sodium alginate...

This study developed novel chitosan (CS)-oxidized sodium alginate (OSA) scaffolds, loaded with formononetin (FMNT), for wound healing applications. Scaffolds were fabricated using 10 % CS and varying OSA concentrations (20 %, 30 %, 50 %). Swelling and degradation studies identified CS-20 %OSA as optimal, exhibiting a moderate 180 % swelling ratio and the slowest degradation rate (65 % weight loss over 7 days), ensuring enhanced stability. Upon incorporating 0.5 mg of FMNT, the CS-20 %OSA-FMNT scaffolds were thoroughly characterized using FTIR, XRD, SEM, TGA, and DSC, confirming structural integrity, thermal stability, and favourable morphology. Biological evaluations demonstrated excellent hemocompatibility with minimal hemolysis (<1 %) and cell viability (>80 %) on HaCaT and NIH/3 T3 cell lines, confirming safety and biocompatibility. Crucially, the scaffolds exhibited significant antibacterial activity. In vitro wound healing assays showed remarkable efficacy, with CS-20 %OSA-FMNT achieving 82.1 % wound coverage at 24 h and near-complete closure by 48 h. In vivo excision wound models in Swiss albino mice further supported these findings, with CS-20 %OSA-FMNT scaffolds achieving faster wound contraction, shorter epithelialization time, and near-complete tissue regeneration compared with controls. Molecular docking and dynamics simulations revealed strong interactions between FMNT and key proteins, such as VEGFR1 and EGFR, elucidating its therapeutic mechanism. In conclusion, the CS-20 %OSA-FMNT scaffold represents a stable, biocompatible, and therapeutically effective biomaterial, paving the way for novel and promising applications in the field of wound healing.