Background: Metal oxide nanoparticles (NPs) have shown promising efficacy for combating bacterial resistance due to their antibacterial properties. This research investigated the effect of zinc oxide NPs (ZnO-NPs) on the antibacterial activity of conventional antibiotics including ciprofloxacin (CIP), cefotaxime (CTX), and colistin (CST) against multidrug-resistant Acinetobacter isolates. Methods: The disc diffusion method was performed to detect the pattern of antibiotic resistance in isolates. The synthesized ZnO-NPs via the solvothermal method were characterized by field emission scanning electron microscopy (FESEM), X-ray diffraction (XRD), and energy-dispersive X-ray spectroscopy (EDS). Finally, the broth microdilution technique was conducted to demonstrate the antibacterial activity of CIP, CTX, and CST antibiotics with and without a sub-inhibitory concentration of ZnO-NPs. Results: XRD, EDS, and FESEM results confirmed the crystalline structure of ZnO-NPs, and the average size was 100±58.68 nm. All isolates were discovered to be of multidrug-resistant (MDR) type and fully susceptible to CST. The antibacterial activity of CTX and CIP was restored when combined with a sub-inhibitory level of ZnO-NPs (0.25 mg/L), and the highest activity was obtained at the concentrations of 32 µg/mL CTX and 8 µg/ mL CIP. Eventually, ZnO-NPs showed a synergistic effect on the antibacterial properties of CST against MDR Acinetobacter. Conclusions: This research indicated that the combination of ZnO-NPs with some common antibiotics can be considered as a novel strategy for reducing the spread of antibiotic-resistant bacteria.