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Introduction: It has been demonstrated that cytokines are important factors in the progression of periodontitis and some of them have an important role in the pathogenesis of periodontal disease. The aim of this study was to compare IL-23 levels in gingival crevicular fluid of patients with chronic periodontitis and healthy subjects. In addition, the relationship between IL-23 levels and probing pocket depths was assessed. Materials & Methods: In this in vitro study, samples were collected from the gingival crevicular fluid of 19 patients with chronic periodontitis and 19 healthy subjects, using paper points. The samples were transferred to the laboratory in transfer vials. IL-23 levels were determined using ELISA reader. The relation between IL-23 levels and probing depths was assessed. Data were analyzed with SPSS 20 using independent t-test and Pearson’s correlation coefficient (α = 0.05). Results: The results showed statistically significant differences in IL-23 levels in GCF between healthy subjects and patients with chronic periodontitis, with higher IL-23 levels in patients with chronic periodontitis (p value < 0.001). In addition, there was a significant and direct correlation between IL-23 levels and pocket depths in patients with chronic periodontitis (p value < 0.001, r = 0.811) and healthy subjects (p value < 0.001, r = 0.858). Conclusion: Based on the results of the present study, IL-23 levels in gingival crevicular fluid can be used for the diagnosis of chronic periodontitis. [ABSTRACT FROM AUTHOR]