A bstract Aim and objective Evaluation of the effectiveness of laser biostimulation (LBS), ice, and local anesthetic (LA) gel in reducing the injection pain during administration of local anesthesia in children. Materials and methods A 3-arm, crossover randomized controlled trial included 30 children of age 9–12 years requiring extraction of primary maxillary posterior teeth. Children were randomly allocated to 3 groups of 10 each. After proper isolation and drying of the buccal mucosa, one of the three techniques, i.e., either LBS or ice or LA gel was applied for 1 minute followed by administration of the LA solution. The pain response was assessed using Wong-Baker Faces Pain Rating Scale and the Sound Eyes Motor scale (SEM). Kruskal–Wallis ANOVA and Mann–Whitney U tests were performed for intragroup and intergroup comparisons, respectively. Results Lower pain score of zero suggesting no hurt was given by more children in the ice group, followed by LA gel and LBS groups in both the scales. The differences in pain scores recorded were found to be statistically significant. Conclusion Ice is found to be equally effective as LA gel, whereas low-level laser therapy is less effective compared to the other two techniques in reducing the injection pain during administration of maxillary posterior buccal infiltration in children. Clinical significance Pain management during LA injection is a critical step in gaining initial trust and during the subsequent treatment visits. The present study suggests that simple methods like pre-cooling the injection site with ice can be used as an effective non-pharmacological technique to reduce injection pain. How to cite this article AmruthaVarshini I, Vinay C, Uloopi KS, et al. Effectiveness of Pre-cooling the Injection Site, Laser Biostimulation, and Topical Local Anesthetic Gel in Reduction of Local Anesthesia Injection Pain in Children. Int J Clin Pediatr Dent 2021;14(1):81–83.