ObjectiveThis randomized controlled clinical trial aimed to evaluate the effect of the casein phosphopeptide-amorphous calcium phosphate fluoride (CPP-ACPF) and photobiomodulation (PBM) in the treatment of dentin hypersensitivity (DH), and the impact of this on the health-related quality of life (HRQL).MethodsEighty teeth with DH were randomized into four groups and received three treatment sessions: PLACEBO = placebo + LASER application mimicking; CPP-ACPF = CPP-ACPF + LASER application mimicking; PBM = placebo + LASER active application; CPP-ACPF+PBM = CPP-ACPF + LASER active application. Tactile (exploratory probe) and evaporative (triple syringe) stimuli were used to measure DH and were recorded with the aid of a visual analogue scale (VAS) after the 1st, 2nd and 3rd treatment sessions and one-month follow-up. The HRQL was recorded in the DH experience questionnaire (DHEQ).ResultsThe intragroup comparison showed a significant reduction in DH (p < 0.05) with both stimuli after one-month follow-up. The intergroup comparison with the evaporative stimulus showed that CPP-ACPF+PBM significantly reduced DH when compared to the rest of treatments, after one-month follow-up. CPP-ACPF+PBM group statistically differed from the other treatment groups in the DHEQ evaluation after one-month follow-up.ConclusionAfter one-month follow-up, the association of CPP-ACPF with PBM was effective in the reduction of DH and promoted a positive impact on the HRQL of the participants of this study.