Since the clock of antimicrobial resistance was set, modern medicine has shed light on a new cornerstone in technology to overcome the worldwide dread of the post-antimicrobial era. Research organizations are exploring the use of nanotechnology to modify metallic crystals from macro to nanoscale size, demonstrating significant interest in the field of antimicrobials. Herein, the antimicrobial activities of aluminum oxide (Al2O3), cobalt aluminum oxide (CoAl2O4), and aluminum doped zinc oxide (Zn0.9Al0.1O) nanoparticles were examined against some nosocomial pathogens. The study confirmed the formation and characterization of Al2O3, CoAl2O4, and Zn0.9Al0.1O nanoparticles using various techniques, revealing the generation of pure nanoscale nanoparticles. With inhibition zones ranging from 9 to 14 mm and minimum inhibitory concentrations varying from 4 mg/mL to 16 mg/mL, the produced nanoparticles showed strong antibacterial activity against Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa, and Staphylococcus aureus. Meanwhile, the bactericidal concentrations ranged from 8 mg/mL to 40 mg/mL. In culture, Zn0.9Al0.1O NPs demonstrated a unique ability to inhibit the development of nosocomial infections with high bactericidal activity (8 mg/mL). Transmission electron microscope images revealed changes in cell shape, bacterial cell wall morphology, cytoplasmic membrane, and protoplasm due to the introduction of tested nanoparticles. These results pave the way for the use of these easily bacterial wall-piercing nanoparticles in combination with potent antibiotics to overcome the majority of bacterial strains' resistance.