Biofilms represent an increasing challenge in the medical practice worldwide, imposing a serious threat to public health. As bacterial strains have developed antibiotic resistance, researcher's attention has been extensively focused on developing more efficient antimicrobial strategies. In this context, the present study reports the synthesis, physicochemical characterization, ex vivo biodistribution, and in vitro evaluation of the capacity of nanostructured surfaces based on zinc oxide (ZnO) and biologically active molecules to modulate clinically relevant microbial biofilms. ZnO nanoparticles (NPs) were synthesized through a co-precipitation method without thermal treatment. The matrix-assisted pulsed laser evaporation (MAPLE) was applied for preparing nanostructured coatings based on ZnO NPs surface modified with linalool that were further characterized by X-ray diffraction (XRD), thermogravimetric analysis with differential scanning calorimetry (TGA-DSC), scanning electron microscopy (SEM), transmission electron microscopy with selected area electron diffraction (TEM-SAED), Fourier-transform infrared spectroscopy (FT-IR), and infrared microscopy (IRM). Histological analyses carried out at 7 days and 14 days after the intraperitoneal administration of linalool modified ZnO NPs revealed the absence of the latter from the brain, kidney, liver, lung, myocardium, and pancreas. Through in vitro assays on prokaryotic cells, it was proven that ZnO coatings hinder microbial biofilm formation of both Gram-positive and Gram-negative bacteria strains.