ISSN : 0376-4672
Purpose : The purpose of this study is to investigate the effects of Simvastatin, which is HMG-CoA reductase inhibitor, on proliferation and differentiation of osteoblast. Materials & Methods : Twenty-four cell culture plates containing essential medium were seeded with UMR-106 cell lines, at density of 5 x 104 cells per plate. Each plates were incubated with 5% CO2 incubator at 37℃. Starting from 2 days after incubation, cell culture medias were replaced with Osteogenesis induction media every 2 days, for 12 days. In some plates, 0.01, 0.1, 1, 10, 100μM of Simvastatin were added with Osteogenesis induction media, and classified as "test group". Those not added with Simvastatin were classified as "control group". Results : 1. When Alrizarin Red staining was observed with naked eye, control group showed normal deep red color, but test group show rapid decrease of red color as Simvastatin concentration increased more than 0.1μM. 2, When observed with microscope, compared to control group, amount of osteo matrix stained with Alrizarin Red decreased rapidly in Simvastatin concentration more than 0.1μM. 3. In optical density analysis, regarding control group as a basis, mineral deposition decreased rapidly when Simvastatin concentration increased more than 0.1μM. 4. In flow cytometry analysis, survival rate of UMR-106 cell showed no changes in both control group and test group. Conclusion : From the above results, we were able to identify that Simvastatin inhibited osteogenesis without effecting survival or cell number of osteoblasts.
1. MERCK & CO., INC. ZORCO (SIMVASTATIN); DISCRIPTION.
2. Hunninghake DB. Therapeutic efficacy of the lipidlowering armamentarium: the clinical benefits of aggressive lipid-lowering therapy. Am J Med 1998;104:9-13.
3. Rogers MJ. Statins: lower lipids and better bones? Nature Medicine 2000;6:21-23.
4. Mundy G, Garrett R, Harris S et al. Stimulation of Bone Formation in Vitro and Rodents by Statins. Science 1999;286:1946-1949.
5. Maeda T, Kawane T, Horiuchi N. Statins Augment Vascular Endothelial Growth Factor Expression in Osteoblastic Cells via Inhibition of Protein Prenylation. Endocrinology 2003;144:681-692.
6. Hwang RJ, Lee EJ, Kim MH et al. Calcyclin, a Ca2+ Ion-binding Protein, Contributes to the Anabolic Effects of Simvastatin on Bone. J Biological Chemistry 2004;279:21239-21247.
7. Edwards CJ, Spector TD. Statin as modulators of bone formation. Arthritis Res 2002;1:151-153.
8. Parhami F, Mody N, Gharavi N et al. Role of Cholesterol Biosynthetic Pathway in Osteoblastic Differentiation of Marrow Stromal Cells. J bone Miner Res 2002;17:1997-2003.
9. Nuckolls GH, Kane A, Shum L, Slavkin HC. Lovastatin promotes cartilage growth but inhibits endochondral ossification of cranial base in organ culture. J Bone Miner Res 2000;15:1(abstract)
10. von Stechow D, Fish S, Yahalom D et al. Does simvastatin stimulate bone formation in vivo? BMC Musculoskeletal Disorder 2003;4:8-17.
11. van Staa TP, Wegman S, de Vries F et al. Use of Statins and Risk of Fractures. JAMA 2001;285:1850-1855.
12. Rejnmark L, Buus NH, Vestergaard P et al. Effects of Simvastatin on Bone Turnover and BMD: A 1-Year Randomized Controlled Trial in Postmenopausal Osteopenic Women. J Bone Miner Res 2004;19:737-744.
13. Wu B, Elmariah S, Kalplan FS et al. Paradoxical Effects of Statins on Aortic Valve Myofibroblasts and Osteoblasts Implication for End-stage Vascular Heart Disease. Arterioscler Thromb Vasc Biol 2005;25:592-597.
14. Beak KH, Lee WY, Oh KW et al. The Effect of Simvastatin on the Proliferation and Differentiation of Human Bone Marrow Stromal Cells. J Korean Med Sci 2005;20:438-444.
15. Maritz FJ, Conradie MM, Hulley PA et al. Effect of statin on Bone Mineral Density and Bone Histomorphometry in Rodents. Arterioscler Thromb Vasc Biol 2001;21:1636-1641.
16. Garrett R, Mundy GR. The role of statins as potential targets for bone formation. Arthritis Res 2002;4:237-240.
17. Choudhury NG, Mandal CC, Choudhury GG. Statin-induced Ras Activation Integrates the Phosphatidylinositol 3-Kinase Signal to Akt and MAPK for Bone Morphogenetic Protein-2 Expression in Osteoblast Differentiation. The Journal of Biological Chemistry 2007;282:4938- 4993.
18. Midy V, Plouet J. Vasculotropin/ vascular endothelial growth factor induces differentiation in cultured osteoblasts. Biochem Biophys Res Commun 1994;199:380-386.
19. Maeda T, Matsunuma A, Kawane T, Horiuchi N. Simvastatin promotes osteoblast differentiation and mineralization in MC3T3-E1 cells. Biochem Biophys Res Commun 2001;280:874-877.
20. Partridge NC, Alcom D et al. Morphological and biochemical characterization of four clonal osteogenic sarcoma cell lines of rat origin. Cancer Research 1983;43:4308-4314.
21. Rajamannan NM, Subramaniam M, Springett M et al. Atorvastatin Inhibits Hypercholersterolemia- Induced Cellular Proliferation and Bone Matrix Production in the Rabbit. Aortic Valve Circulation 2002;105:2660-2665.
22. Novaro GM, Tiong IY, Pearce GL et al. Effect of Hydroxymethylglutaryl Coenzyme A Reductase Inhibitors on the Progression of Calcific Aortic Stenosis. Circulation 2001;104:2205-2209.
23. Takemoto M, Liao JK. Pleiotropic Effects of 3- Hydroxy-3-Methylglutaryl Coenzyme A Reductase Inhibitors. Arterioscler Thromb Vasc Biol 2001;21:1712-1719.
24. Yao W, Farmer R, Cooper R et al. Simvastatin did not prevent nor restore ovariectomy-induced bone loss in adult rats. J Musculoskeletal Neuronal Interact 2006;6(3):277-283.
25. Casey PJ, Seabra MC. Protein prenyltransferases. J Biol Chem 1996;271:5289-5292.
26. Fisher JE, Halasy JM, Masarachia PJ et al. Alendronate mechanism of action: inhibition of osteoclastogenesis and bone resorption are mimicked by lovastatin and blocked by geranylgeraniol. Bone 1998;23:S339.
27. Fisher JE, Rogers MJ, Halasy JM et al. Alendronate mechanism of action: geranylgeraniol, an intermediate in the mevalonate pathway, prevents inhibition of osteoclast formation, bone resorption, and kinase activation in vitro. Cell Biology 1999;96:133-138.
28. Plotkin LI. Prevention of osteocyte and osteoblast apoptosis by bisphosphonates and calcitonin. J Clin Invest 1999;104:1363-1374.
29. Thunyakitpisal PD, Chaisuparat R. Simvastatin, an HMG-CoA Reductase Inhibitor, reduced the Expression of Matrix Metalloproteinase-9 (Gelatinase B) in Osteoblastic Cell and HT1080 Fibrosarcoma Cells. J Pharmacol Sci 2004;94:403- 409.
30. Fromigué O, Haÿ E, Modrowski D et al. RhoA GTPase inactivation by statins induces osteosarcoma cell apoptosis by inhibiting p42/p44- MAPKs-Bcl signaling independently of BMP-2 and cell differentiation. Cell Death and Differentiation 2006;13:1845-1856.
31. Zhong WB, Wang CY, Chang TC, Lee WS. Lovastatin Induces Apoptosis of Anaplastic Thyroid Cancer Cell via Inhibition of Protein Geranylgeranylation and de novo Protein Synthesis. Endocrinology 2003;144:3852-3859.
32. Cafforio P, Dammacco F, Gernone A, Silvestris F. Statins activate the mitochondrial pathway of apoptosis in human lymphoblasts and myeloma cells. Carcinogenesis 2003;26:883-891.