Background: N-terminal-pro-brain natriuretic peptide (NT-proBNP) is an inactive hormone that is seen during inflammation and is a known biomarker of cardiovascular disease (CVD). Evidence suggests that periodontitis has a bidirectional relationship with CVD and NT-proBNP has a potential role in periodontal disease. However, there is no evidence on the impact of nonsurgical periodontal therapy (NSPT) on the levels of NT-proBNP in gingival crevicular fluid (GCF) and serum in patients with chronic periodontitis. Hence, the aim of this study was to compare the levels of NT-proBNP in GCF and serum in patients with chronic generalized periodontitis. Materials and methods: GCF and serum samples were collected in 19 patients with chronic periodontitis before and after NSPT after 6 weeks and the cumulative or reduction in values of NT-proBNP in GCF and serum was assessed. NT-proBNP levels in GCF and serum were determined by enzyme-linked immunosorbent assay. Results: The concentrations of NT-proBNP were significantly reduced in GCF and serum after NSPT. Statistically significant difference of NT-proBNP concentration between pre- and postgroups in GCF was apparent (p < 0.0001), whereas statistically nonsignificant results in NT-proBNP serum levels when compared at baseline to postoperative state with mean of 61.77 (22.6 standard deviation [SD]) preoperatively and 72.67 (51.86 SD) postoperatively (p = 0.0007) was observed. Conclusion: Significant reduction of NT-proBNP concentrations in GCF and serum in patients with chronic periodontitis subjected to NSPT was observed. This may account for a significant relation between periodontal disease, bacteremia, and CVD.