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Background: Very few studies have examined the impact of the concurrent presence of periodontitis (PD) and type 2 diabetes mellitus (T2DM) on serum Vitamin D levels, particularly in developing nations like India, and needs further investigation. Aim: This study aims to assess the relationship between serum Vitamin D values and PD in T2DM patients and to study the correlation between socioeconomic and demographic variables that influence the serum Vitamin D levels and the extent of PD in patients with T2DM. Materials and Methods: This was a cross-sectional, hospital-based research. Medical, dental, and diet histories were obtained from the participants, and their socioeconomic status (SES) was determined. Clinical parameters – plaque index (PI), gingival index (GI), sites with gingival bleeding, probing pocket depth (PPD), and clinical attachment level (CAL) were compared among three groups -patients with generalized Stage III Grade B PD with T2DM (n=35), patients with generalized stage III Grade B PD (n=35) and healthy controls (n=35) and the clinical parameters - plaque index(PI), gingival index(GI), sites with gingival bleeding, probing pocket depth(PPD), and clinical attachment level(CAL) were measured. Biochemical tests included the evaluation of serum 25-hydroxyvitamin D (25[OH] D) and hemoglobin A1C (HbA1c) levels. Statistical Analysis: Periodontal and biochemical parameters were compared using a one-way analysis of variance across the three groups. The association between clinical parameters, SES, and 25(OH)D was examined using Pearson’s correlation coefficient test and linear regression analysis. Results: The serum 25(OH)D levels were lowest in the subjects with generalized Stage III Grade B PD with T2DM (13.54 ± 3.31 ng/mL). Furthermore, there was a significant (P < 0.01) negative correlation between serum 25(OH)D and periodontal parameters, PI (−0.442), PPD (−0.474), CAL (−0.459), sites with gingival bleeding (−0.354), and GI (−0.346) among the groups. The regression analyses showed that an increase in periodontal parameters (PI, GI, PPD, and CAL) and a higher HbA1c was linked to a lower 25(OH)D. However, the periodontal parameters and 25(OH)D levels showed no correlation with socioeconomic and demographic parameters in the study. Conclusion: Serum Vitamin D values are negatively influenced by the synergistic effect of PD and T2DM or by the presence of PD alone. However, the association of SES on serum Vitamin D values in individuals with PD and T2DM or PD alone could not be demonstrated.