Objective To assess the pain profile of patients in the levelling/alignment phase of orthodontic treatment, as reported from randomized clinical trials. Materials and methods Five databases were searched in September 2022 for randomized clinical trials assessing pain during levelling/alignment with a visual analogue scale (VAS). After duplicate study selection, data extraction, and risk-of-bias assessment, random effects meta-analyses of mean differences (MDs) and their 95% confidence intervals (CIs) were performed, followed by subgroup/meta-regression, and certainty analyses. Results A total of 37 randomized trials including 2277 patients (40.3% male; mean age 17.5 years) were identified. Data indicated quick pain initiation after insertion of orthodontic appliances (n = 6; average = 12.4 mm VAS), a quick increase to a peak at day 1 (n = 29; average = 42.4 mm), and gradually daily decrease the first week until its end (n = 23; average = 9.0 mm). Every second patient reported analgesic use at least once this week (n = 8; 54.5%), with peak analgesic use at 6 h post-insertion (n = 2; 62.3%). Patients reported reduced pain in the evening compared to morning (n = 3; MD =  − 3.0 mm; 95%CI =  − 5.3, − 0.6; P = 0.01) and increased pain during chewing (n = 2; MD = 19.2 mm; 95% CI = 7.9, 30.4; P < 0.001) or occlusion of the back teeth (n = 2; MD = 12.4 mm; 95% CI = 1.4, 23.4; P = 0.3), while non-consistent effects were seen for patient age, sex, irregularity, or analgesic use. Subgroup analyses indicated increased pain among extraction cases and during treatment of the lower (rather than the upper) arch, while certainty around estimates was moderate to high. Conclusions Evidence indicated a specific pain profile during orthodontic levelling/alignment, without signs of consistent patient-related influencing factors.