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The capacity of integration of metallic nanoparticles in polymer matrices has improved the antimicrobial behavior, resulting in the search for composites with increased bactericidal properties. A polycaprolactone (PCL) film polymer with copper oxide nanoparticles (CuONPs) was prepared. Dynamic light scattering analysis showed the sizes from 88 to 97 nm of CuONPs. Scanning electron microscopy (SEM) revealed CuONPs with semispherical shapes with diameter 35 nm. The prepared PCL-CuONPs exhibited a nanoporous structure by SEM. The antibacterial applicability of the composite was evaluated to determine the minimum inhibitory concentration in 6 different bacteria and the experimental tests were carried by disk diffusion and spectrophotometric methods. The PCL-CuONPs exhibit a considerable antibacterial effect in gram-positive bacteria in contrast to gram-negative bacteria. 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