Tooth loss exacerbates the deterioration of physical function and induces illness. Numerous studies have identified the risk factors for tooth loss, and several have identified an association of tooth loss with sociodemographic factors, general health status, and lifestyle. The objective of the present cohort study was to elucidate the relationship between regular dental scaling and tooth loss in middle-aged and elderly individuals in Korea. The study was 3-year prospective longitudinal study and conducted in Wonju-si of South Korea. In total, 557 subjects (219 men, 338 women; 40-75 years) were included in our 3-year follow-up survey (2010-2014). Data from the Korean Genome and Epidemiology Study on Atherosclerosis Risk of Rural Areas in the Korean General Population (KOGES-ARIRANG) were used. All subjects underwent an oral examination and face-to-face interview for taking oral health behavior, sociodemographic status, and the utilization of dental service. Logistic regression analysis was used to determine the effects of regular dental scaling on tooth loss after adjusting for history of oral examinations and dental visits, oral health behavior, and sociodemographic status. In total, 263 subjects (47.2%) experienced a loss of one or more teeth during the 3-year period, and lost a mean of 1.54 ± 2.53 teeth. The incidence of tooth loss was 1.87 (1.03-3.38) times higher in participants who did not undergo dental scaling during the 3-year period than in those who regularly received dental scaling. This study showed the potential causal relationship between tooth loss and regular dental scaling for preventing oral disease. Further study is needed to consolidate the evidence that regular dental scaling is effective in preventing tooth loss.