Hydroxyapatite is the main component of enamel that gives the tooth a bright white appearance and eliminates the diffused reflection of light by filling up the fine pores of the tooth surface. Accordingly, remineralization of the teeth can be expected to some extent if hydroxyapatite is used to treat an incipient caries lesion (early white spot lesion). In addition, the remineralization effect will be increased if the particle size of hydroxyapatite can be reduced to less than that of the micron-size in existing toothpaste preparations. The aim of this in vitro study was to evaluate the effect of nano-hydroxyapatite toothpaste, which was produced by nano-technology, on the remineralization of human enamel. A tooth specimen, on which artificial incipient caries had been induced, was immersed into two toothpaste slurries for remineralization. One contains nano-sized hydroxyapatite and fluoride, and the other contains nano-sized hydroxyapatite excluding fluoride. In order to evaluate the remineralization effect, the Vickers Hardness Number & SEM image of the enamel surface was evaluated at each step. There were significant differences in VHN values between those obtained before and after the remineralization steps. The results showed that the remineralization effect increased with increasing immersing time (P<0.05). However, there were no significant differences in VHN values between the two groups (P>0.05). SEM also demonstrated differences the in micro surface at each step. In conclusion, a toothpaste containing nano-sized hydroxyapatite has the potential to remineralize an incipient caries lesion. In addition, the addition of fluoride had no synergistic effect on remineralization.