Effects of Nicotine on Antioxidant Defense Enzymes and RANKL Expression in Human Periodontal Ligament Cells

Background: This study examined the effects of nicotine on osteoblastic differentiation and the osteoclastogenesis regulatory molecules receptor activator of nuclear factor-kappa B ligand (RANKL) and osteoprotegerin (OPG). In addition, we investigated the mechanism by which nicotine induced antioxidant defense enzyme expression as a protective response. Methods: The expression of osteoblast markers, RANKL, OPG, and antioxidant defense enzymes were examined in nicotine-treated human periodontal ligament (PDL) cells by reverse transcription-polymerase chain reaction and Western blotting. Results: Nicotine treatment concomitantly downregulated the expression of OPG and osteoblastic differentiation markers, such as alkaline phosphatase, osteocalcin, and osteopontin, and upregulated the expression of RANKL. Nicotine induced the synthesis of the transcription factor NF-E2-related factor-2 (Nrf2) as well as a number of cellular antioxidants and phase II enzymes, such as heme oxygenase-1. Pretreatment with antioxidants inhibited the upregulation of RANKL, the downregulation of OPG expression, and cytotoxicity by nicotine in PDL cells. Conclusions: Nicotine upregulated RANKL and antioxidant defense enzymes. These data suggest that Nrf2-mediated induction of cellular antioxidants and phase 11 enzymes could contribute to the cellular defense against nicotine-induced cytotoxicity and osteoclastic differentiation in PDL cells. J Periodontol 2009;80:1281-1288.