Nonthermal atmospheric plasma has been studied for its many biomedical effects, such as tooth bleaching, wound healing, and coagulation. In this study, the effects of dentinal tubules occlusion were investigated using fluoride-carboxymethyl cellulose (F-CMC) gel, nano-sized hydroxyapatite (n-HA), and nonthermal atmospheric plasma. Human dentin specimens were divided to 5 groups (group C, HA, HAF, HAP, and HAFP). Group HA was treated with n-HA, group HAF was treated with n-HA after a F-CMC gel application, group HAP was treated with n-HA after a plasma treatment and group HAFP was treated with n-HA after a plasma and F-CMC gel treatment. The occlusion of dentinal tubules was investigated using scanning electron microscopy (SEM) and energy dispersive x-ray spectroscopy (EDS), which shows Ca/P ratio. In the EDS results, a higher Ca/P ratio was shown in the groups including n-HA than in the control group. The specimens of group HAP and HAFP had a higher Ca/P ratio in retentivity. In the SEM results, there was not a significant difference in the amount of times applied. Therefore, this study suggests F-CMC gel and n-HA treatment using nonthermal atmospheric plasma will be a new treatment method for decreasing hypersensitivity.