Resveratrol prevents alveolar bone loss in an experimental rat model of periodontitis
Graphical abstract
Introduction
Periodontitis is an immune-inflammatory disorder that originates due to the formation of complex subgingival microbial biofilms. Periodontitis stimulates inflammatory cells to produce pro-inflammatory cytokines that leads to the destruction of connective tissues such as the subgingivae, periodontal ligament, and alveolar bone [1]. The induction of oxidative stress is a potential mechanism by which periodontitis manifests its systemic effects [2]. Considerable evidence also implicates the association of reactive oxygen species (ROS) with the pathogenesis of periodontitis [3]. Therefore, it is likely that natural compounds having antioxidant potential are capable of ameliorating periodontal damage by inhibiting inflammatory responses and ROS accumulation.
Human gingival fibroblasts (hGFs) play an important role in the local immune response that may initiate periodontitis or gingivitis [4]. The gram-negative bacterium Porphyromonas gingivalis is the main pathogen involved in the initiation and progression of periodontitis [5]. This pathogen produces several virulence factors that stimulate hGFs to produce inflammatory mediators, such as prostaglandin E2, matrix metalloproteinases (MMPs), and several pro-inflammatory cytokines. These mediators lead periodontal cells to mount an excessive host inflammatory response, resulting in periodontal destruction.
Various biological substances are known to have antibacterial activity that promotes the healing and regeneration of periodontal tissues [6], [7]. The use of anti-inflammatory drugs or inhibitors specific to MMPs and pro-inflammatory cytokines are thought to exert beneficial effects on periodontal diseases [8]. However, prolonged exposure to and/or excessive use of antibiotics can contribute to the development of resistance to various antibiotics. The systemic use of several commercial drugs is also known to cause severe side effects leading to patient complications [9]. Accordingly, plant extracts or plant-derived active constituents have been considered as attractive materials to treat periodontitis or repair bone defects [10], [11], as polyphenols isolated from various foods and herbs have been shown to prevent inflammatory diseases [12]. These findings suggest that naturally occurring bioactive substances can act as anti-inflammatory mediators in the process of periodontitis.
Resveratrol is a polyphenolic phytoalexin found in various plants and fruits and is known to exert various pharmacological activities, including antioxidation, anti-inflammation, anticancer, cardioprotection, and vasoprotection [13]. The administration of resveratrol was shown to inhibit the loss of alveolar bone and reduce interleukin (IL)-17 levels in gingival tissue [14]. The compound also activated the nuclear factor erythroid 2-related factor 2 (Nrf2) pathway and diminished oxidative stress and pro-inflammatory cytokine production in a rat periodontitis model [15]. In addition, resveratrol suppressed the inflammatory response of human gingival epithelial cells via the inhibition of nuclear factor (NF)-κB signaling [16]. Furthermore, no toxic effects of the compound were found, even with long-term use [17]. These findings indicate that resveratrol suppresses the inflammatory response in periodontal tissue and improves alveolar bone loss by elevating the expression of cellular redox-sensitive molecules without side effects.
In this study, we employed an experimental periodontitis model using lipopolysaccharide (LPS) and ligature to investigate the protective effect of resveratrol on periodontitic tissue damage. We also examined the mechanisms by which resveratrol prevents the LPS-induced production of inflammatory mediators and ROS in hGFs.
Section snippets
Chemicals and laboratory wares
Resveratrol was obtained from Sigma–Aldrich Co. LLC (St. Louis, MO, USA) and dissolved in dimethylsulfoxide (DMSO) at a concentration of 35 mg/ml as the stock solution. Fetal bovine serum (FBS) and PIK-75, an inhibitor of Nrf2, were purchased from HyClone Laboratories, Inc. (Logan, UT, USA) and Selleck Chemicals (Houston, TX, USA), respectively. LPS produced from P. gingivalis was obtained from InvivoGen (San Diego, CA, USA). Primary antibodies specific for cyclooxygenase-2 (COX-2, BS1076),
Effect of resveratrol on cell viability
We first determined the effect of resveratrol on the viability of hGFs by incubating the cells with various concentrations (0–200 μM) of the compound. Treatment with resveratrol for 24 h at concentrations less than 150 μM did not induce cytotoxicity; rather the compound increased viability of the cells in a dose-dependent manner (Fig. 1A). However, a significant reduction (p < 0.001) in the viability of hGFs was found when the cells were incubated with 200 μM resveratrol for 24 or 48 h (Fig. 1A, B).
Discussion
Cellular ROS production and inflammatory cytokine secretion are increased within periodontal tissues during the immune response to periodontal pathogens. If this condition is prolonged and persistent, periodontal tissue destruction accompanied by oxidative damage occurs. Accordingly, an antioxidant-based therapy has been highlighted as an attractive approach to treat inflammatory periodontal diseases. Here, we used resveratrol as a potential antioxidant based on its protective effect against
Conclusions
Stimulation of hGFs with LPS increases the production of COX-2, MMP-2, MMP-9, and TLR4, the accumulation of intracellular ROS, and the phosphorylation of JNK, p38, and AKT, which are almost completely inhibited by treatment with resveratrol. A LPS-mediated reduction in HO-1 and Nrf2 production in hGFs is also blocked by the addition of resveratrol. Especially, resveratrol treatment itself augments HO-1 induction via the activation of the Nrf2-mediated pathway. Most of the results from in vivo
Acknowledgments
This research was supported by the Basic Science Research Program through the National Research Foundation of Korea (KRF) funded by the Ministry of Science, ICT and Future Planning (NRF-2013R1A2A2A01967207). A grant from the KRF endowed to Dr. Govinda Bhattarai (NRF-2014R1A1A2008488) supported a part of this study.
References (48)
- et al.
Periodontal diseases
Lancet
(2005) - et al.
Alteration of metabolomics profiles by titanium nanoparticles in human gingivitis model
Biomaterials
(2015) - et al.
An anti-inflammatory cell-free collagen/resveratrol scaffold for repairing osteochondrol defects in rabbits
Acta Biomater.
(2014) - et al.
Resveratrol improves oxidative stress and prevents the progression of periodontitis via the activation of the Sirt1/AMPK and the Nrf2/antioxidant defense pathways in a rat periodontitis model
Free Radic. Biol. Med.
(2014) - et al.
Effects of dimethyl sulfoxide on the NLRP3 inflammasome
Immunobiology
(2015) - et al.
Immunolocalization of matrix metalloproteinases-2 and -9 during apical periodontitis development
Arch. Oral Biol.
(2009) - et al.
Circulating matrix metalloproteinase-8 (MMP-8) and MMP-9 are increased in chronic periodontal disease and decrease after non-surgical periodontal therapy
Clin. Chim. Acta
(2009) - et al.
Protective effect of resveratrol against acute lung injury induced by lipopolysaccharide via inhibiting the myd88-dependent toll-like receptor 4 signaling pathway
Mol. Med. Rep.
(2014) - et al.
TRAF6 is functional in inhibition of TLR4-mediated NF-κB activation by resveratrol
J. Nutr. Biochem.
(2013) - et al.
Mitochondrial dysfunction promoted by Porphyromonas gingivalis lipopolysaccharide as a possible link between cardiovascular disease and periodontitis
Free Radic. Biol. Med.
(2011)
Resveratrol prevents high glucose-induced epithelial-mesenchymal transition in renal tubular epithelial cells by inhibiting NADPH oxidase/ROS/ERK pathway
Mol. Cell Endocrinol.
Periodontitis and increase in circulating oxidative stress
Jap. Dent. Sci. Rev.
Resveratrol protects primary rat hepatocytes against oxidative stress damage: activation of the Nrf2 transcription factor and augmented activities of antioxidant enzymes
Eur. J. Pharmacol.
Oxidative stress, systemic inflammation, and severe periodontitis
J. Dent. Res.
Reactive oxygen species: a potential role in the pathogenesis of periodontal diseases
Oral Dis.
Life below the gum line: pathogenic mechanisms of Porphyromonas gingivalis
Microbiol. Mol. Biol. Rev.
Periodontal regeneration by FGF-2 (bFGF) in primate models
J. Dent. Res.
Factors that modulate the effects of bone morphogenetic protein-induced periodontal regeneration: a critical review
J. Periodontol.
Host-mediated resolution of inflammation in periodontal diseases
Periodontology
The coxibs, selective inhibitors of cyclooxygenase-2
N. Engl. J. Med.
A morphometric and histopathologic evaluation of the effects of propolis on alveolar bone loss in experimental periodontitis in rats
J. Periodontol.
Diet and health: what should we eat?
Science
Anti-inflammatory effects of resveratrol: possible role in prevention of age-related cardiovascular disease
Ann. N.Y. Acad. Sci.
Resveratrol decreases periodontal breakdown and modulates local levels of cytokines during periodontitis in rats
J. Periodontol.
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