Abstract Introduction Dentin hypersensitivity is a frequent occurrence in dental practice. It is clinically characterized by acute, short, and temporary pain in response to mechanical, chemical, thermal, or osmotic stimuli resulting from dentin exposure. Objective To compare in vivo the effect of an experimental crystalline bioactive material and nanostructured desensitizing on the pain assessment among patients with cervical dentin hypersensitivity. Material and method Thirty patients were selected for this study, who were randomly assigned to two groups (n=15) in a split-mouth design. Each patient received two treatments: group 1 (fluoride gel and bioactive material) and group 2 (fluoride gel and nanostructured desensitizing). Pain analyses were performed using a visual analogue scale, ranging 0-10. Baseline pain measurement was performed prior to initial treatment (T0) and new measurements were carried out weekly for 3 weeks (T1, T2, and T3) before materials were reapplied. Final pain analysis was performed 3 months after initial treatment (T4). Degree of pain reduction was calculated using the formula T0 - Tperiod after treatment. The data of the pain measurements were analyzed using 2-way repeated measure ANOVA and Tukey’s test (α=0.05). Result Regardless of evaluation period, there was no statistical difference on pain reduction between the treatments (p>0.05). The degree of pain was reduced significantly in each evaluated period for all tested treatments (p<0.05), by approximately three degrees after 3 months. Conclusion The tested desensitizing materials were effective on dentin hypersensitivity reduction.