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Background and Aims: Dental caries is one of the most common infectious diseases which could be prevented in various ways. The aim of the present study was to evaluate the effect of a toothpaste containing different concentrations of nano-hydroxyapatite on the remineralization of incipient caries. Materials and Methods: 9 sound first premolar teeth of maxilla and mandible were sectioned to four pieces. 6 specimens were stored in distilled water as control group. The remaining 30 specimens were demineralized for 72 hours and then their microhardness was measured. 0, 0.5, 1, 2 and 5% wt nano-hydroxyapatite were added to the solution of distilled water and toothpaste. Specimens were divided into 5 groups (N=6) and after demineralization, each group were randomly immersed in the above-mentioned solutions for 15 minutes. Next, specimens were kept in artificial saliva for 1 hour followed by immersion in the related remineralizing solution for another 15 minutes. This procedure was repeated for 5 days. Finally, the microhardness of the teeth was measured. Data were analyzed using repeated measures analysis of variance (ANOVA). Results: The microhardness of demineralized teeth was increased following exposure to different concentrations of nano-hydroxyapatite, but this increase was not statistically significant (P=0.62). Conclusion: Nano-hydroxy apatite can enhance remineralization and increase the tooth microhardness although this increase was not significant.