In this research, as an alternative to chemical and physical methods, environmentally and cost-effective antimicrobial zinc oxide nanoparticles (ZnO NP) were produced by the green synthesis method. The current study focuses on the production of ZnO NP starting from adequate precursor and Zingiber officinale aqueous root extracts (ginger). The produced ZnO NP was loaded into electrospun nanofibers at different concentrations for various tissue engineering applications such as wound dressings. The produced ZnO NPs and ZnO NP-loaded nanofibers were examined by Scanning Electron Microscopy (SEM) for morphological assessments and Fourier-transform infrared spectrum (FT-IR) for chemical assessments. The disc diffusion method was used to test the antimicrobial activity of ZnO NP and ZnO NP-loaded nanofibers against three representatives strains, Escherichia coli (Gram-negative bacteria), Staphylococcus aureus (Gram-positive bacteria), and Candida albicans (fungi) microorganisms. The strength and stretching of the produced fibers were assessed using tensile tests. Since water absorption and weight loss behaviors are very important in tissue engineering applications, swelling and degradation analyses were applied to the produced nanofibers. Finally, the MTT test was applied to analyze biocompatibility. According to the findings, ZnO NP-loaded nanofibers were successfully synthesized using a green precipitation approach and can be employed in tissue engineering applications such as wound dressing.