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. 2014 Aug;49(4):518-26.
doi: 10.1111/jre.12132. Epub 2013 Oct 7.

Simvastatin inhibits lipopolysaccharide-induced osteoclastogenesis and reduces alveolar bone loss in experimental periodontal disease

Affiliations

Simvastatin inhibits lipopolysaccharide-induced osteoclastogenesis and reduces alveolar bone loss in experimental periodontal disease

J Jin et al. J Periodontal Res. 2014 Aug.

Abstract

Background and objective: Statins are inhibitors of 3-hydroxy-3-methylglutaryl coenzyme A reductase and have anti-inflammatory effects independent of cholesterol lowering. Recent clinical studies have indicated that statin intake has a beneficial effect on periodontal disease. However, the underlying mechanisms have not been well understood. In the current study, we employed a rat model with lipopolysaccharide (LPS)-induced periodontal disease and determined the effect of simvastatin, a commonly prescribed statin, on osteoclastogenesis, gingival inflammation and alveolar bone loss.

Material and methods: Sprague-Dawley rats were injected with Aggregatibacter actinomycetemcomitans LPS in periodontal tissue three times per week for 8 wk and part of the rats with LPS injection were also given simvastatin via gavage. After the treatments, the rat maxillae were scanned by microcomputed tomography and the images were analyzed to determine alveolar bone loss. To explore the underlying mechanisms, the effect of simvastatin on osteoclastogenesis and gingival expression of proinflammatory cytokines were also determined by tartrate-resistant acid phosphatase staining and real-time polymerase chain reaction assays, respectively.

Results: Results showed that LPS treatment markedly increased bone loss, but administration of simvastatin significantly alleviated the bone loss. Results also showed that LPS treatment stimulated osteoclastogenesis and the expression of inflammatory cytokines, but simvastatin significantly modulates the stimulatory effect of LPS on osteoclastogenesis and cytokine expression.

Conclusion: This study demonstrated that simvastatin treatment inhibits LPS-induced osteoclastogenesis and gingival inflammation and reduces alveolar bone loss, indicating that the intake of simvastatin may hinder the progression of periodontal disease.

Keywords: alveolar bone; inflammation; lipopolysaccharide; periodontal disease; simvastatin.

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Figures

Figure 1
Figure 1
Inhibition of LPS-induced alveolar bone loss by simvastatin treatment. Rats were treated with phosphate-buffered saline (PBS) (Group 1, n=7), lipopolysaccharide (LPS) (Group 2, n=8), or LPS plus simvastatin (Group 3, n=13) for 8 weeks and the maxillae were scanned by micro computed tomography (microCT) as described in Methods. Representative microCT images from 3 groups (A–C) were shown. The liner distances between the cement-enamel junction (CEJ) to the alveolar bone crest (ABC) were measured as indicated (D). The distances between CEJ and ABC were compared among 3 groups (E) (n=7, 8 and 13 in Group 1, 2, and 3, respectively). The data in E are mean ± standard deviation (SD). The data were from one of two studies with similar results.
Figure 2
Figure 2
Inhibition of lipopolysaccharide (LPS)-induced osteoclastogenesis by simvastatin treatment. The tartate-resistant acid phosphatase (TRAP) staining was performed on bone tissue sections of the maxillae from PBS-treated (A), LPS-treated (B) or LPS plus simvastatin-treated rats (C). Representative tissue sections from 3 groups were shown. The arrows indicate multinucleated osteoclasts. The numbers of multinucleated TRAP-positive osteoclasts were compared between 3 groups (n=5 for each group) (D).
Figure 3
Figure 3
Inhibition of LPS-stimulated IL-6 and matrix metalloproteinase (MMP)-9 expression in periodontal tissue. RNA was isolated from rat periodontal tissue after treatments with vehicle (PBS) (n=7), LPS (n=8) or LPS plus simvastatin (n=13), and real-time PCR was performed to quantify IL-6 (A) and MMP-9 expression (B). The data are mean ± SD of the ratios of IL-6 or MMP-9 vs GAPDH mRNA.
Figure 4
Figure 4
Inhibition of LPS-stimulated MMP-9 protein expression in periodontal tissue. The gingival tissues surrounding the injection sites were collected from rats treated with vehicle PBS (A, n=7), LPS (B, n=8) or LPS plus simvastatin (C, n=13) and sectioned. The sections were immunostained with anti-MMP-9 antibodies and counterstained with hematoxylin. The MMP-9 protein level was quantified as described in Materials and Methods and compared among three groups (D). The horizontal lines indicate the group median levels.

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