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Chronic inflammation has been implicated in the pathogenesis of gestational diabetes mellitus (GDM). Periodontal disease is associated with increased levels of inflammatory mediators and may be a risk factor for GDM. The authors aimed to examine the association between periodontitis and GDM among non-smoking pregnant females.This case-control study included 50 females who were diagnosed with GDM and 50 age- and hospital-matched females without diabetes in Khon Kaen, Thailand. Full-mouth periodontal examinations were performed during pregnancy by two calibrated dentists who were unaware of the case-control status. Periodontitis was defined as ≥1 site with probing depth (PD) ≥5 mm and clinical attachment level (CAL) ≥2 mm at the same site. Serum samples were collected to measure C-reactive protein (CRP), tumor necrosis factor-α, and interleukin-6 levels. Analyses were performed using conditional logistic regression.Fifty percent of the case females had periodontitis compared to 26% of the controls. Females with GDM had significantly higher mean PD and CAL, more sites with bleeding on probing, and increased levels of CRP compared to the controls. Periodontitis was significantly associated with GDM (odds ratio = 3.00, 95% confidence interval = 1.19 to 7.56). The association remained significant with additional adjustment for family history of diabetes, prepregnancy body mass index, and weight gain during pregnancy.The results suggest that periodontitis is associated with GDM. Therefore, clinicians should assess periodontal conditions of pregnant females.