Bacterial infection is a significant barrier to the natural wound healing process. Silver nanoparticles have antibacterial activity through the destruction of DNA and bacterial membranes. In the present study, a green method was introduced to synthesize silver nanoparticles produced by the Schizophyllum commune. The antibacterial mat loaded with 0.5, 1, and 3% (w/w) of bioactive silver nanoparticles were produced in polyvinyl alcohol (PVA) and schizophyllan (SPG) solution (20:80) with electrospun (PVA/SPG). Then the physicochemical properties of silver nanoparticles and the mats PVA/SPG containing silver nanoparticles were investigated. An animal model also examined antimicrobial activity, cytotoxicity, adhesion, and grade II burn ulcers. The results showed that spherical silver nanoparticles with a diameter of about 40 nm were produced and completely scattered on the nanofibers according to TEM images. A non-covalent interaction was revealed between SPG and AgNPs; the nanofibers were hydrophilic. Nanofibers containing silver nanoparticles had intense antimicrobial activity against E. coli and S. aureus. This inhibition increases with increasing concentration. The cytotoxicity and adhesion results showed that the PVA/SPG-AgNPs 1% sample had the best effect on fibroblast cells. The 1% sample was tested for the animal model, and after 14 days, it had a practical effect on the control. At 14 days post-treatment, a skinny epithelial layer was formed and covered the wound area, and these nanofibers were suitable for wound dressing. Consequently, PVA/SPG-AgNPs nanofibers could be a good option to cover burn wounds by speeding up the wound healing process and treatment costs.