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This work focuses on the obtaining and the bactericidal properties study, in vitro, of hybrid films as potential coating materials to inhibit bacteria proliferation. In consequence, hybrid films from nickel oxide (NiO) and nickel ferrite (NiFe2O4) nanoparticles (NPs) embedded in poly-3-hydroxybutyrate (P3HB) were obtained by the solvent casting method. P3HB@NiO and P3HB@NiFe2O4 hybrid films and P3HB film were characterized by X-ray diffraction (XRD), Raman scattering, and scanning electron microscopy (SEM). The XRD of the hybrid films showed that NiO and NiFe2O4 NPs incorporated in the P3HB conserved their nanometric size, and by Energy-dispersive X-ray spectroscopy (EDS) were observed that NPs are homogeneously distributed in the films. The bactericidal effect of the obtained films was evaluated in vitro from the broth surface method against two opportunistic and nosocomial pathogens, Staphylococcus aureus and Pseudomonas aeruginosa. The results showed that P3HB film, P3HB@NiO, and P3HB@NiFe2O4 hybrid films reduced 90%, 98%, and 97% of the growth of S. aureus, respectively. For P. aeruginosa, their growth was reduced by 90%, 94%, and 96%, respectively. In addition, the cytotoxic effect of NiO and NiFe2O4 NPs, as well as P3HB film, and P3HB@NiO, and P3HB@NiFe2O4 hybrid films was evaluated using human skin cells; keratinocytes and fibroblast, being the NPs less cytotoxic than films. Although P3HB is known as a biocompatible polymer, here is demonstrated that in our work conditions, their films have bactericidal properties and are cytotoxic to keratinocytes and fibroblasts, the first barrier of the human skin. However, the P3HB@NiO and P3HB@NiFe2O4 hybrid films synergize the bactericidal effect between the P3HB and the NPs. On the other hand, the NPs decrease the P3HB cytotoxicity to keratinocytes. The methodology used in this work is particularly suitable for producing hybrid films with antibacterial activity against Gram-positive and Gram-negative bacterial strains. [ABSTRACT FROM AUTHOR]