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Two on the apical third (one medial and one distal). One on the middle third (on the center of the root). Laser application was done on 1st day, 3rd, 7th, 14th, 21th, 28th, 35th, 42nd, 49th, 56th day respectively during the canine retraction phase. Distance was measured on 1st day, 35th day and 63rd day. Appliance activation was done on 1st and 35th day. The extent of canine movement was considered buy Ulixertinib as the decrease of the distance between the distal slot of the canine bracket and the mesial opening of the buccal tube of the first molar, measured with a digital electronic caliper (Digimatic Caliper, Mitutoyo, China, with a measuring range of 150 mm, resolution of 0.01 mm and instrumental error of ��0.03 mm) [Figure 4]. Figure 4 Digital electronic caliper The involved area from both groups received radiographic documentation to verify any occurrence of damage to the adjacent periodontal and dental tissues. Conventional intraoral periapicals (IOPA's) were taken on 1st and 63rd day to check qualitatively any damage to the canine root, adjacent alveolar bone, or periodontal tissue. Random selection was done by one operator, who conveyed to the person doing laser application as to which quadrant of the patient belongs to the LG. The distance measurement was done by third operator who GSK3B had no knowledge about which quadrant had been considered in the LG. The two groups were subjected to statistical analysis by the statistician without knowing as to which group was laser and which one control. Hence, it was a triple blind study. The identities of the groups were revealed only after the results of the statistical analysis were obtained. Statistical analysis The comparison of CG and LG with respect to the rate of tooth movement from 1st to 35th day, 35th to 63rd day and from 1st to 63rd day movement was done using unpaired t-test. The mean value of the LG divided by CG was calculated using paired t-test. All the analysis were carried out using Statistical Package for the Social Sciences 2-Methoxyestradiol software version 16(SPSS Inc., Chicago, IL, USA) (P