Previous studies have reported an association between tooth loss and gray matter volume (GMV) in healthy adults. The study aims to elucidate the link between tooth loss, brain volume differences, and cognitive impairment by investigating the total and regional differences in GMV associated with tooth loss in older people with and without cognitive impairment. Forty older participants with mild cognitive impairment or Alzheimer's disease [the cognitive impairment (CI) group] and 30 age- and sex-matched healthy participants [the control (CON) group] received T1-weighted magnetic resonance imaging scans and assessments of oral functions, including masticatory performance (MP) and the number of missing teeth (NMT). Voxel-based morphometry was used to assess the total and regional GMV, including that of the medial temporal lobe and motor-related areas. (A) When the total intracranial volume and age were controlled for, an increased MP was associated with a larger GMV in the premotor cortex in the CON group. (B) In the CI group, an increased NMT was significantly correlated with smaller regional GMV of the bilateral primary motor cortex and the premotor cortex. (C) In the CI group, but not the CON group, an increased NMT was associated with both smaller total GMV and regional GMV of the left medial temporal lobe, including the left hippocampus and parahippocampus. Tooth loss may be preferentially related to the structural differences in the medial temporal lobe in cognitively impaired older people. Further research is required to understand the mechanisms of the relationships.