Many papers have been published on surgical mandibulectomy and reconstruction. However, only a few reports refer to masticatory function after prosthodontic treatment in mandibulectomy patients. The aim of this study was to investigate the masticatory function of mandibulectomy patients. Twenty-three subjects (10 males and 13 females, with an average age of 63 years) participated in this study: 11 subjects who had undergone unilateral marginal mandibulectomy, six subjects with unilateral segmental mandibulectomy with reconstruction and six subjects with hemimandibulectomy without reconstruction. Mixing Ability Index (MAI) was used to measure masticatory function on the non-defect side and on the defect side with a prosthesis installed. Comparisons were carried out among the marginal, segmental and hemimandibular groups and between the non-defect side and the defect side. Consequently, our study indicates these results. On the non-defect side, a significant difference was found between the marginal and the segmental groups, and between the marginal and the hemimandibular groups. In the marginal and the segmental groups, a significant difference was found between the non-defect and the defect sides. In conclusion, our study suggests that MAI is an adequate tool to study the masticatory function in mandibulectomy patients, the masticatory function of the mandibulectomy patients is more impaired than that of the ordinary removable partial denture patients, and that surgical intervention affects the masticatory function on not only the defect side but also the non-defect side in mandibulectomy patients.