The goal of this study is to improve the qualities of Saudi cotton fabrics to be used in a variety of industrial and medicinal applications. Cotton fabrics were treated by dipping in chitosan/ZnO-NPs solutions with various ZnO-NPs concentrations. XRD and SEM of crystal and morphological structure, thermal analysis (TG and DSC) of fabric stability for heat assessment, were used to characterize the fabrics. The produced textiles were tested for antimicrobial activity against Gram-positive Staphylococcus aureus, Gram-negative Escherichia coli (bacteria), and Aspergillus niger, Candida albicans (fungus) using agar diffusion method. According to the XRD data, the fabrics treated with chitosan and by increasing the concentration of ZnO-NPs had increased crystallinity. The successful deposition and strong binding of chitosan and ZnO-NPs to the fabric surface, and the obvious change in fabric morphology, were confirmed by SEM micrographs. The pyrolysis peak positions were changed to a higher temperature. The creation of the inhibitory zone indicated that the treated fabrics had bactericidal effects. The results could be useful in an open innovation approach to improving the qualities of treated textiles that point to their use in medical applications.