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Undesired biofilm development is a major concern in many areas, especially in the medical field. The purpose of the present study was to comparatively investigate the antibiofilm efficacy of usnic acid, in soluble versus nanofluid formulation, in order to highlight the potential use of Fe3O4/oleic acid (FeOA) nanofluid as potential controlled release vehicle of this antibiofilm agent. The (+) -UA loaded into nanofluid exhibited an improved antibiofilm effect on S. aureus biofilm formation, revealed by the drastic decrease of the viable cell counts as well as by confocal laser scanning microscopy images. Our results demonstrate that FeOA nanoparticles could be used as successful coating agents for obtaining antibiofilm pellicles on different medical devices, opening a new perspective for obtaining new antimicrobial and antibiofilm surfaces, based on hybrid functionalized nanostructured biomaterials.