The aim of this study was to assess the accuracy of 3D rendering of the mandibular condylar region obtained from different semi-automatic segmentation methodology. A total of 10 Cone beam computed tomography (CBCT) were selected to perform semi-automatic segmentation of the condyles by using three free-source software (Invesalius, version 3.0.0, Centro de Tecnologia da Informação Renato Archer, Campinas, SP, Brazil; ITK-Snap, version2.2.0; Slicer 3D, version 4.10.2) and one commercially available software Dolphin 3D (Dolphin Imaging, version 11.0, Chatsworth, CA, USA). The same models were also manually segmented (Mimics, version 17.01, Materialise, Leuven, Belgium) and set as ground truth. The accuracy of semi-automatic segmentation was evaluated by (1) comparing the volume of each semi-automatic 3D rendered condylar model with that obtained with manual segmentation, (2) deviation analysis of each 3D rendered mandibular models with those obtained from manual segmentation. No significant differences were found in the volumetric dimensions of the condylar models among the tested software (p > 0.05). However, the color-coded map showed underestimation of the condylar models obtained with ITK-Snap and Slicer 3D, and overestimation with Dolphin 3D and Invesalius. Excellent reliability was found for both intra-observer and inter-observer readings. Despite the excellent reliability, the present findings suggest that data of condylar morphology obtained with semi-automatic segmentation should be taken with caution when an accurate definition of condylar boundaries is required.