Modern-day treatment demands scarless wound healing utilizing scaffolds in the form of nanofiber mats which are tissue and environment-friendly. Neem gum polysaccharide (NGP) in conjugation with Polyvinyl alcohol (PVA) in the form of nanofibers exhibits antimicrobial properties mimicking extracellular matrix for tissue growth. Different grades of nanofibers mats (NFM) were prepared by combining different ratios of NGP and PVA which were later crosslinked using glutaraldehyde vapors (25 % w/v in 0.5 M HCl), and optimized grade G14 exhibited maximum tensile strength with smooth surface morphology, hemocompatible properties, in-vitro biodegradability and antimicrobial action against S. aureus &E. coli. G14 was analytically characterized using different analytical techniques viz. Fourier-transform infrared spectroscopy (FTIR), Thermogravimetric analysis (TGA), which indicated polymer-polymer compatibility. The surface hydrophobicity as detected using Optical contact angle (OCA) confirmed the hydrophobicity of NFM with increased glutaraldehyde vapor for crosslinking when compared to non-crosslinked NFM. Histopathology slides indicated G14 CL-NFM accelerated the wound healing in mice with dense collagen and fibroblasts when compared to control mice suggesting the tissue engineering potential of the prepared device.