Background: Nanotechnology represents a new science that promises to provide a broad range of uses and improved technologies for biological and biomedical applications. One of the reasons behind the intense interest is that nanotechnology permits synthesis of materials that have structure is less than 100 nanometers. The present work revealed the effect of zinc oxide nanoparticles (ZnO NPs) on Streptococcus mutans of Human Saliva in comparison to de-ionized water. Materials and methods: Streptococcus mutans were isolated from saliva of forty eight volunteers of both sexes their age range between 18-22 years and then purified and diagnosed according to morphological characteristic and biochemical tests. Different concentrations of ZnO NPs were prepared from the stock solution; all the experiments were conducted in vitro. Disk diffusion method was used to study the sensitivity of Streptococcus mutans to different concentrations of zinc oxide nanoparticles in comparison to effect of de-ionized water. Results: Streptococcus mutans were sensitive to concentrations (0.05, 0.1, 0.5, 1, 3 and 5.8) mg / ml of the zinc oxide nanoparticles solution in comparison to de-ionized water, revealing a highly significant difference in all concentrations except for concentration (0.01) mg / ml which was showed no significant difference in comparison to de-ionized water. Conclusion: This study revealed that zinc oxide nanoparticles were effective against Streptococcus mutans.