Background: The treatment of transversal maxillary deficiency usually aims at skeletal expansion. The treatment option highly depends on the maturation stage of the midpalatal suture (MPS), which may vary between individuals at the same chronological age. Therefore, the individual determination of the MPS maturation is crucial. Aims: Our aim was to investigate the applicability of fractal analysis for the quantitative evaluation of MPS maturation. Methods: Nine experienced orthodontists were asked to evaluate the MPS maturation on 51 cone beam computed tomography (CBCT) scans according to the Angelieri classification method. Intra- and inter-examiner reliability was measured using Cohen’s Kappa coefficient. The stages were agreed upon according to the results of the examiners with the highest strength of agreement. Fractal analysis was then performed on the CBCTs and the correlation between the fractal dimension values and maturation stages was then evaluated. Optimal fractal dimension cut-off values were determined using a receiver operating characteristic curve. Results: The cut-off point was found at 1.073, at which the use of fractal dimension for predicting MPS maturation showed 100% sensitivity, 93.7% specificity, 9.5% false positive, 0% false negative rate. Conclusion: Our results provided further evidence that fractal analysis is a reliable tool to determine MPS maturation stage.