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The effects of dexamethasone on the apoptosis and osteogenic differentiation of human periodontal ligament cells

  • Kim, Sung-Mi (Department of Periodontology, Kyungpook National University School of Dentistry) ;
  • Kim, Yong-Gun (Department of Periodontology, Kyungpook National University School of Dentistry) ;
  • Park, Jin-Woo (Department of Periodontology, Kyungpook National University School of Dentistry) ;
  • Lee, Jae-Mok (Department of Periodontology, Kyungpook National University School of Dentistry) ;
  • Suh, Jo-Young (Department of Periodontology, Kyungpook National University School of Dentistry)
  • Received : 2013.02.05
  • Accepted : 2013.06.14
  • Published : 2013.08.31

Abstract

Purpose: The purpose of the current study was to examine the effect of dexamethasone (Dex) at various concentrations on the apoptosis and mineralization of human periodontal ligament (hPDL) cells. Methods: hPDL cells were obtained from the mid-third of premolars extracted for orthodontic reasons, and a primary culture of hPDL cells was prepared using an explant technique. Groups of cells were divided according to the concentration of Dex (0, 1, 10, 100, and 1,000 nM). A 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay was performed for evaluation of cellular viability, and alkaline phosphatase activity was examined for osteogenic differentiation of hPDL cells. Alizarin Red S staining was performed for observation of mineralization, and real-time polymerase chain reaction was performed for the evaluation of related genes. Results: Increasing the Dex concentration was found to reduce cellular viability, with an increase in alkaline phosphatase activity and mineralization. Within the range of Dex concentrations tested in this study, 100 nM of Dex was found to promote the most vigorous differentiation and mineralization of hPDL cells. Dex-induced osteogenic differentiation and mineralization was accompanied by an increase in the level of osteogenic and apoptosis-related genes and a reduction in the level of antiapoptotic genes. The decrease in hPDL cellular viability by glucocorticoid may be explained in part by the increased prevalence of cell apoptosis, as demonstrated by BAX expression and decreased expression of the antiapoptotic gene, Bcl-2. Conclusions: An increase in hPDL cell differentiation rather than cellular viability at an early stage is likely to be a key factor in glucocorticoid induced mineralization. In addition, apoptosis might play an important role in Dex-induced tissue regeneration; however, further study is needed for investigation of the precise mechanism.

Keywords

References

  1. Seo BM, Miura M, Gronthos S, Bartold PM, Batouli S, Brahim J, et al. Investigation of multipotent postnatal stem cells from human periodontal ligament. Lancet 2004;364: 149-55. https://doi.org/10.1016/S0140-6736(04)16627-0
  2. Bartold PM, Shi S, Gronthos S. Stem cells and periodontal regeneration. Periodontol 2000 2006;40:164-72. https://doi.org/10.1111/j.1600-0757.2005.00139.x
  3. Gould TR, Melcher AH, Brunette DM. Migration and division of progenitor cell populations in periodontal ligament after wounding. J Periodontal Res 1980;15:20-42. https://doi.org/10.1111/j.1600-0765.1980.tb00258.x
  4. McCulloch CA, Bordin S. Role of fibroblast subpopulations in periodontal physiology and pathology. J Periodontal Res 1991;26(3 Pt 1):144-54. https://doi.org/10.1111/j.1600-0765.1991.tb01638.x
  5. Chung CH, Golub EE, Forbes E, Tokuoka T, Shapiro IM. Mechanism of action of beta-glycerophosphate on bone cell mineralization. Calcif Tissue Int 1992;51:305-11. https://doi.org/10.1007/BF00334492
  6. Kodama HA, Amagai Y, Sudo H. Culture conditions affecting differentiation and calcification in the MC3T3-E1 osteogenic cell line. In: Ali SY, editor. Cell-mediated calcification and matrix vesicles. Amsterdam: Elsevier Science BV; 2006. p.297-302.
  7. Franceschi RT, Iyer BS, Cui Y. Effects of ascorbic acid on collagen matrix formation and osteoblast differentiation in murine MC3T3-E1 cells. J Bone Miner Res 1994;9:843-54.
  8. Majeska RJ, Nair BC, Rodan GA. Glucocorticoid regulation of alkaline phosphatase in the osteoblastic osteosarcoma cell line ROS 17/2.8. Endocrinology 1985;116:170-9. https://doi.org/10.1210/endo-116-1-170
  9. Leboy PS, Beresford JN, Devlin C, Owen ME. Dexamethasone induction of osteoblast mRNAs in rat marrow stromal cell cultures. J Cell Physiol 1991;146:370-8. https://doi.org/10.1002/jcp.1041460306
  10. Shalhoub V, Conlon D, Tassinari M, Quinn C, Partridge N, Stein GS, et al. Glucocorticoids promote development of the osteoblast phenotype by selectively modulating expression of cell growth and differentiation associated genes. J Cell Biochem 1992;50:425-40. https://doi.org/10.1002/jcb.240500411
  11. Abe Y, Aida Y, Abe T, Hirofuji T, Anan H, Maeda K. Development of mineralized nodules in fetal rat mandibular osteogenic precursor cells: requirement for dexamethasone but not for beta-glycerophosphate. Calcif Tissue Int 2000;66:66-9. https://doi.org/10.1007/s002230050013
  12. Bellows CG, Heersche JN, Aubin JE. Determination of the capacity for proliferation and differentiation of osteoprogenitor cells in the presence and absence of dexamethasone. Dev Biol 1990;140:132-8. https://doi.org/10.1016/0012-1606(90)90060-V
  13. Wang H, Pang B, Li Y, Zhu D, Pang T, Liu Y. Dexamethasone has variable effects on mesenchymal stromal cells. Cytotherapy 2012;14:423-30. https://doi.org/10.3109/14653249.2011.652735
  14. Weinstein RS, Jilka RL, Parfitt AM, Manolagas SC. Inhibition of osteoblastogenesis and promotion of apoptosis of osteoblasts and osteocytes by glucocorticoids: potential mechanisms of their deleterious effects on bone. J Clin Invest 1998;102:274-82. https://doi.org/10.1172/JCI2799
  15. Parfitt AM. Bone-forming cells in clinical conditions. In: Hall BK, editor. Bone. Caldwell: Telford Press and CRC Press; 1990. p.351-429.
  16. Jilka RL, Weinstein RS, Bellido T, Parfitt AM, Manolagas SC. Osteoblast programmed cell death (apoptosis): modulation by growth factors and cytokines. J Bone Miner Res 1998;13:793-802. https://doi.org/10.1359/jbmr.1998.13.5.793
  17. Proudfoot D, Skepper JN, Hegyi L, Bennett MR, Shanahan CM, Weissberg PL. Apoptosis regulates human vascular calcification in vitro: evidence for initiation of vascular calcification by apoptotic bodies. Circ Res 2000;87:1055-62. https://doi.org/10.1161/01.RES.87.11.1055
  18. Shin JH, Park JW, Yeo SI, Noh WC, Kim MK, Kim JC, et al. Identification of matrix mineralization-related genes in human periodontal ligament cells using cDNA microarray. J Korean Acad Periodontol 2007;37(Suppl):447-63.
  19. Kim HS, Park JW, Yeo SI, Choi BJ, Suh JY. Effects of high glucose on cellular activity of periodontal ligament cells in vitro. Diabetes Res Clin Pract 2006;74:41-7. https://doi.org/10.1016/j.diabres.2006.03.034
  20. Bligh ME, Bhagwat SA, Castonguay TW. Aldosterone diurnal rhythm in the rat: a question of cross-reactivity? Physiol Behav 1993;53:845-8. https://doi.org/10.1016/0031-9384(93)90260-M
  21. Malendowicz LK, Nussdorfer GG, Markowska A, Tortorella C, Nowak M, Warchol JB. Effects of neuromedin U (NMU)-8 on the rat hypothalamo-pituitary-adrenal axis. Evidence of a direct effect of NMU-8 on the adrenal gland. Neuropeptides 1994;26:47-53.
  22. Fleshner M, Deak T, Spencer RL, Laudenslager ML, Watkins LR, Maier SF. A long-term increase in basal levels of corticosterone and a decrease in corticosteroid-binding globulin after acute stressor exposure. Endocrinology 1995; 136:5336-42. https://doi.org/10.1210/endo.136.12.7588279
  23. Sharma AC, Bosmann HB, Motew SJ, Hales KH, Hales DB, Ferguson JL. Steroid hormone alterations following induction of chronic intraperitoneal sepsis in male rats. Shock 1996;6:150-4. https://doi.org/10.1097/00024382-199608000-00011
  24. Ishida Y, Bellows CG, Tertinegg I, Heersche JN. Progesterone-mediated stimulation of osteoprogenitor proliferation and differentiation in cell populations derived from adult or fetal rat bone tissue depends on the serum component of the culture media. Osteoporos Int 1997;7:323-30. https://doi.org/10.1007/BF01623772
  25. Tenenbaum HC, Heersche JN. Differentiation of osteoblasts and formation of mineralized bone in vitro. Calcif Tissue Int 1982;34:76-9. https://doi.org/10.1007/BF02411212
  26. Canalis E, Delany AM. Mechanisms of glucocorticoid action in bone. Ann N Y Acad Sci 2002;966:73-81. https://doi.org/10.1111/j.1749-6632.2002.tb04204.x
  27. Beloti MM, Rosa AL. Osteoblast differentiation of human bone marrow cells under continuous and discontinuous treatment with dexamethasone. Braz Dent J 2005;16:156-61. https://doi.org/10.1590/S0103-64402005000200013
  28. McCulloch CA, Tenenbaum HC. Dexamethasone induces proliferation and terminal differentiation of osteogenic cells in tissue culture. Anat Rec 1986;215:397-402. https://doi.org/10.1002/ar.1092150410
  29. Shen Q, Zhu S, Hu J, Geng N, Zou S. Recombinant human bone morphogenetic protein-4 (BMP-4)-stimulated cell differentiation and bone formation within the expanding calvarial suture in rats. J Craniofac Surg 2009;20:1561-5. https://doi.org/10.1097/SCS.0b013e3181b09cc1
  30. Choi MH, Noh WC, Park JW, Lee JM, Suh JY. Gene expression pattern during osteogenic differentiation of human periodontal ligament cells in vitro. J Periodontal Implant Sci 2011;41:167-75. https://doi.org/10.5051/jpis.2011.41.4.167
  31. Cheng SL, Zhang SF, Avioli LV. Expression of bone matrix proteins during dexamethasone-induced mineralization of human bone marrow stromal cells. J Cell Biochem 1996;61:182-93. https://doi.org/10.1002/(SICI)1097-4644(19960501)61:2<182::AID-JCB3>3.0.CO;2-Q
  32. Hayami T, Zhang Q, Kapila Y, Kapila S. Dexamethasone's enhancement of osteoblastic markers in human periodontal ligament cells is associated with inhibition of collagenase expression. Bone 2007;40:93-104. https://doi.org/10.1016/j.bone.2006.07.003
  33. Stein GS, Lian JB, Owen TA. Relationship of cell growth to the regulation of tissue-specific gene expression during osteoblast differentiation. FASEB J 1990;4:3111-23. https://doi.org/10.1096/fasebj.4.13.2210157
  34. Porter RM, Huckle WR, Goldstein AS. Effect of dexamethasone withdrawal on osteoblastic differentiation of bone marrow stromal cells. J Cell Biochem 2003;90:13-22. https://doi.org/10.1002/jcb.10592
  35. Nakashima K, Zhou X, Kunkel G, Zhang Z, Deng JM, Behringer RR, et al. The novel zinc finger-containing transcription factor osterix is required for osteoblast differentiation and bone formation. Cell 2002;108:17-29. https://doi.org/10.1016/S0092-8674(01)00622-5
  36. Lynch MP, Capparelli C, Stein JL, Stein GS, Lian JB. Apoptosis during bone-like tissue development in vitro. J Cell Biochem 1998;68:31-49. https://doi.org/10.1002/(SICI)1097-4644(19980101)68:1<31::AID-JCB4>3.0.CO;2-X
  37. Ducy P, Geoffroy V, Karsenty G. Study of osteoblast-specific expression of one mouse osteocalcin gene: characterization of the factor binding to OSE2. Connect Tissue Res 1996;35:7-14. https://doi.org/10.3109/03008209609029169
  38. Dejean LM, Martinez-Caballero S, Manon S, Kinnally KW. Regulation of the mitochondrial apoptosis-induced channel, MAC, by BCL-2 family proteins. Biochim Biophys Acta 2006;1762:191-201. https://doi.org/10.1016/j.bbadis.2005.07.002
  39. Seo MS, Hwang KG, Kim H, Baek SH. Analysis of gene expression during odontogenic differentiation of cultured human dental pulp cells. Restor Dent Endod 2012;37:142-8. https://doi.org/10.5395/rde.2012.37.3.142
  40. Fujita H, Yamamoto M, Ogino T, Kobuchi H, Ohmoto N, Aoyama E, et al. Necrotic and apoptotic cells serve as nuclei for calcification on osteoblastic differentiation of human mesenchymal stem cells in vitro. Cell Biochem Funct 2013 May 8 [Epub]. http://dx.doi.org/doi:10.1002/cbf.2974.

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