Background <p>Pain control during needle insertion reduces stress and increases collaboration between the patient and the dentist. The aim of this study was to compare the effect of photobiostimulation, pressure, and topical anesthetic gel on reducing pain during infiltration injection.</p> Materials &amp; methods <p>A participant- and operator-blinded crossover clinical trial was conducted using 16 healthy volunteers ranging from 20 to 40 years of age. Each of the four interventions was done for each person at intervals of one week: (1) Photobiostimulation (photobiomodulation therapy, PBMT) using low-power diode laser with 808&#xa0;nm, 400 mW continuous wave at an applied energy density of 2&#xa0;J/cm². (2) 20% benzocaine topical anesthetic gel. (3) Pressure and (4) Control group. Then injection of 0.6 mL 2% lidocaine into the buccal mucosa of the maxillary canine was done. All subjects recorded pain using the visual analog scale (VAS). The Friedman and Dunn tests were used for data analysis.</p> Results <p>The mean scores of VAS for the laser, gel, pressure, and control groups were 1.43, 2.90, 1.78, and 3.15, respectively. The mean pain level in the laser group (<i>p</i> = 0.02) and pressure (<i>p</i> = 0.03) was lower than the control group.</p> Conclusion <p>Within the limitations of this small split-mouth trial, photobiomodulation therapy (PBMT) and local pressure appear to reduce pain during maxillary infiltration injection, whereas 20% benzocaine gel did not differ significantly from the control.</p> Trial registration <p>Iranian Registry of Clinical Trials, IRCT20141222020396N2; registered 21 January 2018.</p>