The periapical local anesthetic injection may be associated with fear of needles and pain administration. Dental topical anesthetic agents can help to reduce pain perception; however, adverse events can occur. To investigate the efficacy of 1064-nm photobiomodualtion (PBM) in inducing mucosal anesthesia delivered with a flat-top hand-piece compared to 20% Benzocaine topical anesthetic gel, sixty healthy patients were randomly allocated (1:1) to either 20% benzocaine topical gel + placebo laser (T group) or PBM + placebo gel (L group). The 1064-nm Nd:YAG laser was employed and is associated with a novel flat-top hand piece. The applied operational parameters were 0.5 W, 10 Hz, 100 µs pulse width, and 30 J/cm2 for one-minute single application time. The enrolled subjects were asked to assess pain intensity at the time of anesthetic injection with a Visual Analog Scale. Taking into consideration taste, undesirable numbness, and overall satisfaction, the patients were asked to rate their experiences according to a verbal rating scale. Statistical analysis showed no statistically significant difference between the T and L Groups for pain ratings (p = 0.0596). The L Group displayed significantly higher ratings than T Group for taste, undesirable numbness, and overall satisfaction (p < 0.001). The 1064-nm PBM delivered by flat-top hand piece is effective in inducing mucosal anesthesia, eliminating the adverse side-effects of the conventional topical anesthetic gel.