This study is related to the biosynthesis of copper nanoparticles using plant extracts of Ficus religiosa (peepal vriksh) and its potential as an effective antimicrobial agent, with special reference to wound-associated pathogens that are reported to form biofilms in chronic wounds. Wound biofilm-forming microorganisms hinder the healing process, and the clinical samples taken from wounds were processed. The identification and pathogenicity of the isolated microbes were tested using biochemical and hemolysis tests. The biosynthesis of copper nanoparticles was performed using Ficus religiosa plant leaf extract and was characterized using UV-visible, XRD, and FTIR techniques, and their efficacy was evaluated using the well-diffusion method. According to the observed hemolytic patterns, the microbes isolated from the wound samples were pathogenic.