Attention on phyto-synthesized nanoparticles (NPs) has recently increased due to their low chemical toxicity, which gaining significance in nanotechnology. Our research synergizes ginger extract and nickel oxide NPs to make a new composite of antibacterial potential against bacterial strains, namely: P. aeruginosa, S. aureus, E. coli, and K. pneumonia. The prepared NPs were filled into Polymethacrylic acid (PMMA) films, where they displayed notable bactericidal effects, particularly against S. aureus and E. coli. Three examination techniques were applied to characterize the produced sheets, namely: scanning electron microscopy, atomic force microscopy, and X-ray diffraction. Simultaneously, NiO nanoparticles induced the generation of reactive oxygen species, disrupting bacterial functions, and altering cellular morphology. This study highlights the need for continuous research into effective and ecologically friendly antimicrobial treatments.