Multidrug resistance (MDR) infectious wounds are a major concern due to drug resistance, leading to increased patient morbidity. Lichenysin (LCN), a lipopeptide and biosurfactant obtained from certain strains of Bacillus licheniformis, has demonstrated an excellent antimicrobial property. The present study focuses on the fabrication and comprehensive evaluation of LCN-incorporated poly(vinyl alcohol) (PVA)/polycaprolactone (PCL)-based nanofiber scaffolds using an electrospinning technique as a potential wound healing biomaterial for the treatment of MDR infectious wounds in diabetic rats. The LCN-loaded PVA-PCL nanofiber scaffolds were characterized for their physicochemical, antimicrobial, in vitro cell line on L-929, hemocompatibility, flow cytometry, in vivo infectious wound healing, and enzyme-linked immuno sorbent assay (ELISA). Morphological analysis via scanning electron microscopy (SEM) images confirmed smooth and porous nanofibers with diameters in the range 200-300 nm. Fourier transform infrared and X-ray diffraction (XRD) results demonstrated the structural integrity, chemical compatibility, and amorphous nature of developed scaffolds. The scaffolds loaded with LCN demonstrated excellent water retention, moderate biodegradability, and sustained release of LCN for up to 72 h. Mechanical characterization demonstrated a robust tensile strength conducive to wound healing applications. Antimicrobial activity against Pseudomonas aeruginosa (P. aeruginosa) and Staphylococcus aureus (S. aureus) showed substantial antibacterial and antibiofilm activity. In vitro cell line studies showed enhanced cell adhesion, proliferation, migration, and viability, signifying the cytocompatibility of these scaffolds. In vivo studies demonstrated exceptional infectious wound healing potential in diabetic rats. These findings indicate that LCN-enriched PVA-PCL scaffolds hold significant potential as a therapeutic strategy for the treatment of MDR infectious wounds in diabetic rats through a multifaceted approach.