Introduction : Many studies have aimed to investigate the regeneration potential of bone substitutes through animal models at different defect sites, where the bone healing mechanism varies due to developmental, structural and functional differences. This study aims to develop a rabbit model with two functionally different (non-load-bearing calvarias and load-bearing mandibular) critical-sized defects (CSD) in one rabbit. Material & Method : The comparison of the “gold standard” autograft to a sham (no graft) control was undertaken in order to validate this model; at the same time, a 3D-printed biphasic calcium phosphate scaffold was implanted to test its utility in the evaluation of new bone substitute materials. Twenty rabbits were selected with both a 10 mm calvaria defect and a 11 mm bicortical semi-cylindrical mandibular defect. The animals were euthanized at 4 and 12 weeks once surgery, microcomputed tomography and histological analysis had been performed. Results : In the case of the calvaria, the results for the non-healing sham group compared with the healing of those that had undergone the autograft validated the CSD model. But the mandibular defect was not validated, due to the particularity of mandible high mechanical stress and infectious risk. Discussion : This study showed for the first time that rabbits have a high tolerance for the bilateral double-site CSD model under consideration; and further studies are essential to modify and improve the design of mandibular CSD.