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The antimicrobial properties of two-dimensional materials such as graphene-based surfaces are vital for environmental and biomedical applications. Here, the improvement of the antibacterial property of reduced graphene oxide by the preparation of rGO-CuO nanocomposite films was reported. The rGO-CuO nanocomposites were synthesized via a simple hydrothermal method, and the nanocomposite films were fabricated by filtering through a polytetrafluoroethylene (PTFE) filter with the assistance of a vacuum filtration unit. After characterization of the nanocomposite films, the antibacterial properties were tested against Pseudomonas aeruginosa PAO1. The fabricated rGO-CuO nanocomposite films exhibited excellent antibacterial activity, leading to complete bacterial inactivation upon contact. The antibacterial properties were closely linked to the reactive oxygen species (ROS) independent pathway rather than …