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The packaging system is one of the most challenging researches, in which antibacterial agents were embedded. In this research activity, we used folic acid (FA), as a biosafe and biodegradable substance for the surface modification of zinc oxide nanoparticles (ZnO NPs). Then, the ZnO-FA NPs as nanofillers were inserted in the poly(vinyl alcohol) (PVA) with different weight percentages (2, 5, and 8 wt%, relative to polymer weight). The synthesized substances were characterized using various analyses like Ultraviolet-Visible (UV–Vis) spectroscopy, water contact angle, field emission scanning electron microscopy, and thermogravimetric analysis. The image of transmission electron microscopy analysis displayed the excellent dispersal of the ZnO-FA NPs within the polymeric matrix. By increasing the percentage of the modified ZnO NPs into the PVA, the intensity of the absorption peaks in the UV–Vis spectra was raised, and the thermal stability of NC films got better compared to the pure polymer. PVA/ZnO-FA NC film 8 wt% showed better antibacterial activity than the pure PVA and had the best resistance against Gram-positive bacteria (Staphylococcus aureus).

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