Nanoparticles of zinc oxide (ZnO) were added to poly(ε-caprolactone) (PCL), and PCL/ZnO casting films were produced, afterwards films were characterized using Fourier Transform Infrared Spectroscopy (FTIR), x-ray Diffraction (XRD), Atomic Force Microscopy (AFM) and Scanning Electron Microscopy (SEM). Additionally, antimicrobial and cytotoxic parameters were determined using microbial adhesion testing according to JIS Z 2801: 2000 (E) and agar diffusion method according to ISO 10993-5 2009, respectively. From collected data, the chemical identity of individual components was kept and the surface control could be achieved changing the composition. According to FTIR spectra and using the Lambert-Beer law higher interaction ratios were met for higher ZnO content which is linked to antimicrobial action. As revealed by AFM analysis, at 5% of ZnO, nanoparticles were well dispersed in PCL matrix with uniform surface film. Analyses from antibacterial activity and cytotoxicity suggested pathogenic Staphylococcus aureus growth was hindered in ZnO films; specifically for PCL/ZnO 5% optimal antibacterial activity and toxicity absent were reached. Summing up, PCL/ZnO5% nanocomposites films offer great potential for commercial applications as active food packaging.