Background: The present study aimed to investigate the effects of low-level laser therapy (LLLT) on orthodontic tooth movement and its correlation with the levels of interleukin-1 beta (IL-1 beta), receptor activator of nuclear factor kappa B ligand (RANKL) and osteoprotegerin (OPG) in gingival crevicular fluid (GCF). Methods: This split-mouth design study included 12 patients scheduled for the extraction of both upper first premolars. Patients were randomly selected for experimental group that received left- or right-side radiation with a diode laser (810 nm wavelength, 100 mW power output, 6.29 J/cm(2) energy density). Laser treatment was applied on days 0, 7, 14, and 21, after loading the canine retraction forces. GCF concentrations of IL-1 beta, RANKL, and OPG were analyzed. The upper arch of each patient was scanned with an intraoral scanner to assess tooth movement. Results: The cumulative tooth movement over 28 days was significantly higher in the laser group than in the control group. We observed significant reductions in OPG levels and increases in IL-1 beta and RANKL levels in GCF samples on the experimental sides. Conclusion: With the parameter settings used in this study, LLLT could, to some extent, lead to changes in bone metabolism, which could accelerate orthodontic tooth movement.