New green synthesis of Ag-nanoparticles (Ag-NPs) using O-Carboxymethyl Chitosan (O-CMCs) as stabilizing agent and ascorbic acid as reducing agent was achieved. The reaction was carried out in an autoclave at a pressure of 0.12 MPa and a temperature of 120 degrees C with varying concentrations of solution precursors and different reaction times. The size, shape and structure of Ag-NPs were measured using Transmission Electron Microscope (TEM), X-Ray Diffraction (XRD), FT-IR and UV Spectrophotometers. The Ag-NPs stabilized in O-CMCs were blended with polyvinyl alcohol (PVA) polymer solution and then electrospun to produce nanofibers with high antimicrobial activity. The morphological study of O-CMCs/PVA/Ag-NPs nanofibers was characterized using SEM. Finally, the release behavior of Ag-NPs from these nanofibers was examined and the antimicrobial activity was measured against some skin pathogenic bacteria and fungi using the agar diffusion method. The newly developed membranes show a unique antimicrobial activity against the tested strains and were presented as promising active wound dressing materials in medical applications.