Visfatin is an adipocytokine that plays a role in regulating periodontal inflammation by as yet identified mechanisms. It has been suggested that visfatin mediates inflammation via activation of the nuclear factor-kappa B (NF-kappa B) and phosphatidylinositol 3-kinase (PI3k) signaling pathways which play a role in the inhibition of neutrophil apoptosis. The aim of this study was to investigate the expression of visfatin, NF-kappa B (NF-kappa B-1 and NF-kappa B-2), PI3k, tumor necrosis factor alpha (TNF-alpha), and interleukin-1 beta (IL-1 beta) in the tissue of healthy individuals and patients with periodontitis. Tissue biopsy samples were obtained from 21 patients with chronic periodontitis and from the gingiva of 19 healthy individuals undergoing crown lengthening. The mRNA expression levels of visfatin, NF-kappa B, PI3k, TNF-alpha, and IL-1 beta were evaluated by quantitative real-time PCR (qPCR). Also, visfatin protein expression was measured by immunohistochemistry. Both qPCR and immunohistochemistry results revealed that the visfatin expression was higher in the tissues with periodontitis than in healthy tissues (P < 0.01). Similarly, the mRNA expression levels of NF-kappa B-2, PI3k, and IL-1 beta were higher in tissues with periodontitis than in healthy gingival tissues (P < 0.01). Visfatin was positively correlated with the levels of NF-kappa B-1 (r = 0.549, P < 0.05), NF-kappa B-2 (r = 0.636, P < 0.05), PI3k (r = 0.682, P < 0.01), TNF-alpha (r = 0.558, P < 0.05), and IL-1 beta (r = 0.686, P < 0.01) in the tissues with periodontitis. Our results demonstrated that increased visfatin was associated with the expression of NF-kappa B and PI3k which may play a role in the pathogenesis of periodontitis. We suggest that increased visfatin may contribute to the inhibition of neutrophil apoptosis via the NF-kappa B and PI3k signaling pathways. Understanding the role of visfatin in periodontitis will enable the development of new treatment methods for inflammation.