This work presents a novel nano-modified coating for wound dressings and other medical devices with anti-infective properties, based on functionalized zinc oxide nanostructures and orange oil (ZnO@OO). The obtained nanosurfaces were characterized by transmission electron microscopy (TEM), scanning electron microscopy (SEM), selected area electron diffraction (SAED), differential thermal analysis-thermogravimetry (DTA-TG), X-ray diffraction (XRD), and Fourier transform infrared (FT-IR) spectroscopy. The obtained nanocomposite coatings exhibited an antimicrobial activity superior to bare ZnO nanoparticles (NPs) and to the control antibiotic against Staphylococcus aureus and Escherichia coli, as revealed by the lower minimal inhibitory concentration values. For the quantitative measurement of biofilm-embedded microbial cells, a culture-based, viable cell count method was used. The coated wound dressings proved to be more resistant to S. aureus microbial colonization and biofilm formation compared to the uncoated controls. These results, correlated with the good in vivo biodistribution open new directions for the design of nanostructured bioactive coating and surfaces, which can find applications in the medical field, for obtaining improved bioactive wound dressings and prosthetic devices, but also in food packaging and cosmetic industry.