this study aimed to clarify how masticatory muscle atrophy induced by botulinum toxin (BtX) injection affects cortical bone quality of the mandible using 3D modeling technology. A total of 39 young (26.9 ± 6.0 years) and 38 post-menopausal (55.3 ± 6.3 years) females were included. Computed tomography (CT) images were obtained before and after 12 months of treatment. Predictor variables were application of a stabilization splint, and/or two times of BtX injection in the bilateral temporalis and masseter muscles within a six-month interval. Outcome variables were changes in average Hounsfield units (HU) and cortical thickness of region of interest (ROI). 3D mandibular models were reconstructed using CT images, and models were used to calculate average HU and cortical thickness of Rois, including inferior half of the lateral surface of ascending ramus, coronoid process, and temporomandibular joint condyle. Cortical bone quality at muscle insertion site was influenced by decreased muscle thickness but seemed not to be affected by decreased functional loading. Reduced functional loading seemed to influence cortical bone quality of the condyles. These effects were more remarkable in post-menopausal females. Hence, decreased masticatory muscle thickness may lead to alterations of the mandibular cortical structures, especially in post-menopausal females. Bone is a dynamic structure that continuously undergoes a remodeling process through resorption and apposition to meet environmental requirements. The adaptation rate of the bone to functional loading is mediated by dynamic forces of the associated muscles 1. Skeletal muscles transduce mechanical loading to the bone in three ways: (1) the tensile force developed by a muscle contraction at its insertion site, (2) the compressive force developed by muscles that transduce loading across the joints, and (3) the bending force experienced by long bone associated muscles for lifting distally held objects 2. The role of mechanical stimulation by muscles to maintain bone density has been well known and the association among the amount of physical activity, body muscle mass, and quality of both cortical and trabecular bones has also been investigated 3-6. Botulinum toxin (BTX) acts as a pre-synaptic neurotoxin that blocks neuromuscular transmission by inhibiting acetylcholine release from the motor and sympathetic nerve terminals 7 as well as inflammatory mediators including substance P and glutamate 7,8. Muscle paralysis induced by BTX injection was related with not only reduced active loading but also an increased passive elastic modulus of muscle fiber bundles 9. BTX has been used not only for cosmetic purposes but also to improve symptoms associated with temporomandibular disorders (TMDs), myofascial pain syndrome, and headaches. The majorities of adverse effects associated with the application of BTX in the orofacial region are mild and transient including bruising, swelling, pain around the injection site, and masticatory muscle weakness 10. Several previous studies have rev...