Advertisement

Human Cell

, Volume 25, Issue 2, pp 29–35 | Cite as

Effect of simvastatin on the osteogenetic behavior of alveolar osteoblasts and periodontal ligament cells

  • Shutai Liu
  • Kristina Bertl
  • Hongchen Sun
  • Zhong-Hao Liu
  • Oleh AndrukhovEmail author
  • Xiaohui Rausch-FanEmail author
Research Article

Abstract

Statins are routinely used in the clinic as cholesterol lowering drugs, but recently they were reported to also have anabolic effects on bone tissue. Since regeneration of alveolar bone is one of the primary aims of periodontal treatment, in the present study we investigated the effects of simvastatin, a lipophilic statin, on primary alveolar osteoblasts (AOBs) and periodontal ligament cells (PDLs) in vitro. The effect of simvastatin (1–100 nM) on the cells proliferation/viability after 24, 48, and 72 h stimulation was measured using 3,4,5-dimethylthiazol-2-yl-2,5-diphenyltetrazolium bromide (MTT)-assay. The alkaline phosphatase (ALP) activity was measured after stimulation with simvastatin using specific colorimetric assay. Finally, the mRNA expression levels of osteocalcin (OC), receptor activator of NF-κB ligand (RANKL), and osteoprotegerin (OPG) were measured by real-time PCR. The proliferation/viability of AOBs was significantly decreased by all simvastatin concentrations after 72 h stimulation. The proliferation/viability of PDLs was not influenced by simvastatin. ALP activity of AOBs and PDLs was increased by 1 and 100 nM simvastatin, respectively. Simvastatin induced a dose-dependent increase in OC mRNA expression of AOBs and did not influence that in PDLs. RANKL expression of AOBs was increased at all tested simvastatin concentrations and that in PDLs was increased by higher simvastatin concentrations (10–100 nM). Finally, the expression of OPG in AOBs and PDLs was stimulated by 1–10 and 100 nM simvastatin, respectively. Simvastatin seems to slightly increase the expression of osteogenic markers in AOBs and PDLs, indicating its ability to influence alveolar bone formation and periodontal regeneration.

Keywords

Periodontium Alkaline phosphatase Osteoprotegerin RANK ligand Simvastatin 

Notes

Acknowledgments

We thank Mrs. Nguyen Phuong Quynh, Department of Periodontology, Bernhard Gottlieb Clinic of Dentistry for her fantastic help in preparing and performing the experiments.

Conflict of interest

None.

References

  1. 1.
    Ward S, Lloyd Jones M, Pandor A, et al. A systematic review and economic evaluation of statins for the prevention of coronary events. Health Technol Assess. 2007;11:1–160, iii–iv.Google Scholar
  2. 2.
    Ogura T, Tanaka Y, Nakata T, Namikawa T, Kataoka H, Ohtsubo Y. Simvastatin reduces insulin-like growth factor-1 signaling in differentiating C2C12 mouse myoblast cells in an HMG-CoA reductase inhibition-independent manner. J Toxicol Sci. 2007;32:57–67.PubMedCrossRefGoogle Scholar
  3. 3.
    Jadhav SB, Jain GK. Statins and osteoporosis: new role for old drugs. J Pharm Pharmacol. 2006;58:3–18.PubMedCrossRefGoogle Scholar
  4. 4.
    Garrett IR, Gutierrez G, Mundy GR. Statins and bone formation. Curr Pharm Des. 2001;7:715–36.PubMedCrossRefGoogle Scholar
  5. 5.
    Meier CR, Schlienger RG, Kraenzlin ME, Schlegel B, Jick H. HMG-CoA reductase inhibitors and the risk of fractures. JAMA. 2000;283:3205–10.PubMedCrossRefGoogle Scholar
  6. 6.
    Edwards CJ, Hart DJ, Spector TD. Oral statins and increased bone-mineral density in postmenopausal women. Lancet. 2000;355:2218–9.PubMedCrossRefGoogle Scholar
  7. 7.
    Mundy G, Garrett R, Harris S, et al. Stimulation of bone formation in vitro and in rodents by statins. Science. 1999;286:1946–9.PubMedCrossRefGoogle Scholar
  8. 8.
    Maeda T, Matsunuma A, Kawane T, Horiuchi N. Simvastatin promotes osteoblast differentiation and mineralization in MC3T3–E1 cells. Biochem Biophys Res Commun. 2001;280:874–7.PubMedCrossRefGoogle Scholar
  9. 9.
    Anbinder AL, Junqueira JC, Mancini MN, Balducci I, Rocha RF, Carvalho YR. Influence of simvastatin on bone regeneration of tibial defects and blood cholesterol level in rats. Braz Dent J. 2006;17:267–73.PubMedCrossRefGoogle Scholar
  10. 10.
    Bjarnason NH, Riis BJ, Christiansen C. The effect of fluvastatin on parameters of bone remodeling. Osteoporos Int. 2001;12:380–4.PubMedCrossRefGoogle Scholar
  11. 11.
    LaCroix AZ, Cauley JA, Pettinger M, et al. Statin use, clinical fracture, and bone density in postmenopausal women: results from the Women’s Health Initiative Observational Study. Ann Intern Med. 2003;139:97–104.PubMedGoogle Scholar
  12. 12.
    von Stechow D, Fish S, Yahalom D, et al. Does simvastatin stimulate bone formation in vivo? BMC Musculoskelet Disord. 2003;4:8.CrossRefGoogle Scholar
  13. 13.
    Sodek J, McKee MD. Molecular and cellular biology of alveolar bone. Periodontology. 2000;24:99–126.CrossRefGoogle Scholar
  14. 14.
    Beertsen W, McCulloch CA, Sodek J. The periodontal ligament: a unique, multifunctional connective tissue. Periodontology. 1997;13:20–40.CrossRefGoogle Scholar
  15. 15.
    McCulloch CA, Bordin S. Role of fibroblast subpopulations in periodontal physiology and pathology. J Periodontal Res. 1991;26:144–54.PubMedCrossRefGoogle Scholar
  16. 16.
    Meyer JR. The regenerative potential of the periodontal ligament. J Prosth Dent. 1986;55:260–5.CrossRefGoogle Scholar
  17. 17.
    Gorustovich AA, Steimetz T, Giglio MJ, Guglielmotti MB. A histomorphometric study of alveolar bone modeling and remodeling under experimental anaemia and polycythaemia in rats. Arch Oral Biol. 2006;51:246–51.PubMedCrossRefGoogle Scholar
  18. 18.
    Hasegawa T, Yoshimura Y, Kikuiri T, et al. Expression of receptor activator of NF-kappa B ligand and osteoprotegerin in culture of human periodontal ligament cells. J Periodontal Res. 2002;37:405–11.PubMedCrossRefGoogle Scholar
  19. 19.
    Ozec I, Kilic E, Gumus C, Goze F. Effect of local simvastatin application on mandibular defects. J Craniofac Surg. 2007;18:546–50.PubMedCrossRefGoogle Scholar
  20. 20.
    Stein D, Lee Y, Schmid MJ, et al. Local simvastatin effects on mandibular bone growth and inflammation. J Periodontol. 2005;76:1861–70.PubMedCrossRefGoogle Scholar
  21. 21.
    Yazawa H, Zimmermann B, Asami Y, Bernimoulin JP. Simvastatin promotes cell metabolism, proliferation, and osteoblastic differentiation in human periodontal ligament cells. J Periodontol. 2005;76:295–302.PubMedCrossRefGoogle Scholar
  22. 22.
    Qu Z, Rausch-Fan X, Wieland M, Matejka M, Schedle A. The initial attachment and subsequent behavior regulation of osteoblasts by dental implant surface modification. J Biomed Mat Res. 2007;82:658–68.CrossRefGoogle Scholar
  23. 23.
    Rausch-fan X, Qu Z, Wieland M, Matejka M, Schedle A. Differentiation and cytokine synthesis of human alveolar osteoblasts compared to osteoblast-like cells (MG63) in response to titanium surfaces. Dent Mater. 2008;24:102–10.PubMedCrossRefGoogle Scholar
  24. 24.
    Qu Z, Laky M, Ulm C, et al. Effect of Emdogain on proliferation and migration of different periodontal tissue-associated cells. Oral Surg Oral Med Oral Path Oral Radiol Endod. 2010;109:924–31.CrossRefGoogle Scholar
  25. 25.
    Zhang Y, Schedle A, Matejka M, Rausch-Fan X, Andrukhov O. The proliferation and differentiation of osteoblasts in co-culture with human umbilical vein endothelial cells: an improved analysis using fluorescence-activated cell sorting. Cell Mol Biol Lett. 2010;15:517–29.PubMedCrossRefGoogle Scholar
  26. 26.
    Mundy GR. Statins and their potential for osteoporosis. Bone. 2001;29:495–7.PubMedCrossRefGoogle Scholar
  27. 27.
    Uzzan B, Cohen R, Nicolas P, Cucherat M, Perret GY. Effects of statins on bone mineral density: a meta-analysis of clinical studies. Bone. 2007;40:1581–7.PubMedCrossRefGoogle Scholar
  28. 28.
    Lindy O, Suomalainen K, Makela M, Lindy S. Statin use is associated with fewer periodontal lesions: a retrospective study. BMC Oral Health. 2008;8:16.PubMedCrossRefGoogle Scholar
  29. 29.
    Sakoda K, Yamamoto M, Negishi Y, Liao JK, Node K, Izumi Y. Simvastatin decreases IL-6 and IL-8 production in epithelial cells. J Dent Res. 2006;85:520–3.PubMedCrossRefGoogle Scholar
  30. 30.
    Yamashita M, Otsuka F, Mukai T, et al. Simvastatin antagonizes tumor necrosis factor-alpha inhibition of bone morphogenetic proteins-2-induced osteoblast differentiation by regulating Smad signaling and Ras/Rho-mitogen-activated protein kinase pathway. J Endocrinol. 2008;196:601–13.PubMedCrossRefGoogle Scholar
  31. 31.
    Ruiz-Gaspa S, Nogues X, Enjuanes A, et al. Simvastatin and atorvastatin enhance gene expression of collagen type 1 and osteocalcin in primary human osteoblasts and MG-63 cultures. J Cell Biochem. 2007;101:1430–8.PubMedCrossRefGoogle Scholar
  32. 32.
    Lima LL, Goncalves PF, Sallum EA, Casati MZ, Nociti FH Jr. Guided tissue regeneration may modulate gene expression in periodontal intrabony defects: a human study. J Periodontal Res. 2008;43:459–64.PubMedCrossRefGoogle Scholar
  33. 33.
    Wakabayashi S, Tsutsumimoto T, Kawasaki S, Kinoshita T, Horiuchi H, Takaoka K. Involvement of phosphodiesterase isozymes in osteoblastic differentiation. J Bone Miner Res. 2002;17:249–56.PubMedCrossRefGoogle Scholar
  34. 34.
    Lian JB, Stein GS. Concepts of osteoblast growth and differentiation: basis for modulation of bone cell development and tissue formation. Crit Rev Oral Biol Med. 1992;3:269–305.PubMedGoogle Scholar
  35. 35.
    Khosla S. Minireview: the OPG/RANKL/RANK system. Endocrinology. 2001;142:5050–5.PubMedCrossRefGoogle Scholar
  36. 36.
    Wu B, Elmariah S, Kaplan FS, Cheng G, Mohler ER 3rd. Paradoxical effects of statins on aortic valve myofibroblasts and osteoblasts: implications for end-stage valvular heart disease. Arterioscler Thromb Vasc Biol. 2005;25:592–7.PubMedCrossRefGoogle Scholar
  37. 37.
    Maeda T, Matsunuma A, Kurahashi I, Yanagawa T, Yoshida H, Horiuchi N. Induction of osteoblast differentiation indices by statins in MC3T3–E1 cells. J Cell Biochem. 2004;92:458–71.PubMedCrossRefGoogle Scholar
  38. 38.
    Kim IS, Jeong BC, Kim OS, et al. Lactone form 3-hydroxy-3-methylglutaryl-coenzyme A reductase inhibitors (statins) stimulate the osteoblastic differentiation of mouse periodontal ligament cells via the ERK pathway. J Periodontal Res. 2011;46:204–13.PubMedCrossRefGoogle Scholar
  39. 39.
    Kaji H, Kanatani M, Sugimoto T, Chihara K. Statins modulate the levels of osteoprotegerin/receptor activator of NFkappaB ligand mRNA in mouse bone-cell cultures. Horm Metab Res. 2005;37:589–92.PubMedCrossRefGoogle Scholar
  40. 40.
    Yang H, Feng Y, Luan Y. Determination of simvastatin in human plasma by liquid chromatography-mass spectrometry. J Chromatogr. 2003;785:369–75.CrossRefGoogle Scholar

Copyright information

© Japan Human Cell Society and Springer 2011

Authors and Affiliations

  1. 1.Department of Periodontology, Bernhard Gottlieb University School of DentistryMedical University of ViennaViennaAustria
  2. 2.Yantai Stomatological HospitalBinzhou Medical UniversityYantaiChina
  3. 3.Department of Oral Pathology, School of StomatologyJilin UninversityChangchunChina

Personalised recommendations