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Gene expression pattern during osteogenic differentiation of human periodontal ligament cells in vitro

  • Choi, Mi-Hye (Department of Periodontology, Kyungpook National University School of Dentistry) ;
  • Noh, Woo-Chang (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 : 2011.05.17
  • Accepted : 2011.06.18
  • Published : 2011.08.31

Abstract

Purpose: Periodontal ligament (PDL) cell differentiation into osteoblasts is important in bone formation. Bone formation is a complex biological process and involves several tightly regulated gene expression patterns of bone-related proteins. The expression patterns of bone related proteins are regulated in a temporal manner both in vivo and in vitro. The aim of this study was to observe the gene expression profile in PDL cell proliferation, differentiation, and mineralization in vitro. Methods: PDL cells were grown until confluence, which were then designated as day 0, and nodule formation was induced by the addition of 50 ${\mu}g$/mL ascorbic acid, 10 mM ${\beta}$-glycerophosphate, and 100 nM dexamethasone to the medium. The dishes were stained with Alizarin Red S on days 1, 7, 14, and 21. Real-time polymerase chain reaction was performed for the detection of various genes on days 0, 1, 7, 14, and 21. Results: On day 0 with a confluent monolayer, in the active proliferative stage, c-myc gene expression was observed at its maximal level. On day 7 with a multilayer, alkaline phosphatase, bone morphogenetic protein (BMP)-2, and BMP-4 gene expression had increased and this was followed by maximal expression of osteocalcin on day 14 with the initiation of nodule mineralization. In relationship to apoptosis, c-fos gene expression peaked on day 21 and was characterized by the post-mineralization stage. Here, various genes were regulated in a temporal manner during PDL fibroblast proliferation, extracellular matrix maturation, and mineralization. The gene expression pattern was similar. Conclusions: We can speculate that the gene expression pattern occurs during PDL cell proliferation, differentiation, and mineralization. On the basis of these results, it might be possible to understand the various factors that influence PDL cell proliferation, extracellular matrix maturation, and mineralization with regard to gene expression patterns.

Keywords

Acknowledgement

Supported by : KOSEF

References

  1. Strutz F, Okada H, Lo CW, Danoff T, Carone RL, Tomaszewski JE, et al. Identification and characterization of a fibroblast marker: FSP1. J Cell Biol 1995;130:393-405. https://doi.org/10.1083/jcb.130.2.393
  2. Lackler KP, Cochran DL, Hoang AM, Takacs V, Oates TW. Development of an in vitro wound healing model for periodontal cells. J Periodontol 2000;71:226-37. https://doi.org/10.1902/jop.2000.71.2.226
  3. Nojima N, Kobayashi M, Shionome M, Takahashi N, Suda T, Hasegawa K. Fibroblastic cells derived from bovine periodontal ligaments have the phenotypes of osteoblasts. J Periodontal Res 1990;25:179-85. https://doi.org/10.1111/j.1600-0765.1990.tb01041.x
  4. Somerman MJ, Young MF, Foster RA, Moehring JM, Imm G, Sauk JJ. Characteristics of human periodontal ligament cells in vitro. Arch Oral Biol 1990;35:241-7. https://doi.org/10.1016/0003-9969(90)90062-F
  5. Cho MI, Matsuda N, Lin WL, Moshier A, Ramakrishnan PR. In vitro formation of mineralized nodules by periodontal ligament cells from the rat. Calcif Tissue Int 1992; 50:459-67. https://doi.org/10.1007/BF00296778
  6. Mukai M, Yoshimine Y, Akamine A, Maeda K. Bone-like nodules formed in vitro by rat periodontal ligament cells. Cell Tissue Res 1993;271:453-60. https://doi.org/10.1007/BF02913727
  7. Chung HB, Park JW, Suh JY. The effect of dexamethasone on the gene expression of the bone matrix protein in the periodontal ligament cells. J Korean Acad Periodontol 2002; 32:445-56. https://doi.org/10.5051/jkape.2002.32.3.445
  8. Stein GS, Lian JB. Molecular mechanisms mediating proliferation/differentiation interrelationships during progressive development of the osteoblast phenotype. Endocr Rev 1993;14:424-42.
  9. Choi JY, Lee BH, Song KB, Park RW, Kim IS, Sohn KY, et al. Expression patterns of bone-related proteins during osteoblastic differentiation in MC3T3-E1 cells. J Cell Biochem 1996;61:609-18. https://doi.org/10.1002/(SICI)1097-4644(19960616)61:4<609::AID-JCB15>3.0.CO;2-A
  10. Zhumabayeva BD, Lin WL, Choung PH, Chien HH, Sodek J, Sampath KT, et al. Differential induction of bone sialoprotein by dexamethasone and osteogenic protein-1 (OP-1, BMP-7) in rat periodontal ligament cells in vitro: relationship to the mineralization of tissue nodules. Int J Oral Biol. 1998;91-101.
  11. Strayhorn CL, Garrett JS, Dunn RL, Benedict JJ, Somerman MJ. Growth factors regulate expression of osteoblast-associated genes. J Periodontol 1999;70:1345-54. https://doi.org/10.1902/jop.1999.70.11.1345
  12. 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.
  13. Ripamonti U. Induction of cementogenesis and periodontal ligament regeneration by bone morphogenetic proteins. In: Lindholm TS, editor. Bone morphogenetic proteins: biology, biochemistry and reconstructive surgery. San Diego: Academic Press; 1996. p.189-98.
  14. King GN, King N, Cruchley AT, Wozney JM, Hughes FJ. Recombinant human bone morphogenetic protein-2 promotes wound healing in rat periodontal fenestration defects. J Dent Res 1997;76:1460-70. https://doi.org/10.1177/00220345970760080801
  15. Ivanovski S, Li H, Haase HR, Bartold PM. Expression of bone associated macromolecules by gingival and periodontal ligament fibroblasts. J Periodontal Res 2001;36: 131-41. https://doi.org/10.1034/j.1600-0765.2001.360301.x
  16. Shalhoub V, Gerstenfeld LC, Collart D, Lian JB, Stein GS. Downregulation of cell growth and cell cycle regulated genes during chick osteoblast differentiation with the reciprocal expression of histone gene variants. Biochemistry 1989;28:5318-22. https://doi.org/10.1021/bi00439a002
  17. Preston GA, Lyon TT, Yin Y, Lang JE, Solomon G, Annab L, et al. Induction of apoptosis by c-Fos protein. Mol Cell Biol 1996;16:211-8. https://doi.org/10.1128/MCB.16.1.211
  18. McCabe LR, Banerjee C, Kundu R, Harrison RJ, Dobner PR, Stein JL, et al. Developmental expression and activities of specific fos and jun proteins are functionally related to osteoblast maturation: role of Fra-2 and Jun D during differentiation. Endocrinology 1996;137:4398-408. https://doi.org/10.1210/en.137.10.4398
  19. Aubin JE, Turken K, Heersche JNM. Osteoblastic cell lineage. In: Noda M, editor. Cellular and molecular biology of bone. San Diego: Academic Press; 1993. p.1-44.
  20. Onyia JE, Hale LV, Miles RR, Cain RL, Tu Y, Hulman JF, et al. Molecular characterization of gene expression changes in ROS 17/2.8 cells cultured in diffusion chambers in vivo. Calcif Tissue Int 1999;65:133-8. https://doi.org/10.1007/s002239900671
  21. 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
  22. Hughes FJ, Collyer J, Stanfield M, Goodman SA. The effects of bone morphogenetic protein-2, -4, and -6 on differentiation of rat osteoblast cells in vitro. Endocrinology 1995;136:2671-7. https://doi.org/10.1210/en.136.6.2671
  23. Zegzula HD, Buck DC, Brekke J, Wozney JM, Hollinger JO. Bone formation with use of rhBMP-2 (recombinant human bone morphogenetic protein-2). J Bone Joint Surg Am 1997;79:1778-90.
  24. Thesleff I, Vaahtokari A, Kettunen P, Aberg T. Epithelial-mesenchymal signaling during tooth development. Connect Tissue Res 1995;32:9-15. https://doi.org/10.3109/03008209509013700
  25. Boden SD, McCuaig K, Hair G, Racine M, Titus L, Wozney JM, et al. Differential effects and glucocorticoid potentiation of bone morphogenetic protein action during rat osteoblast differentiation in vitro. Endocrinology 1996;137: 3401-7. https://doi.org/10.1210/en.137.8.3401
  26. Kobayashi M, Takiguchi T, Suzuki R, Yamaguchi A, Deguchi K, Shionome M, et al. Recombinant human bone morphogenetic protein-2 stimulates osteoblastic differentiation in cells isolated from human periodontal ligament. J Dent Res 1999;78:1624-33. https://doi.org/10.1177/00220345990780100701
  27. Zaman KU, Sugaya T, Kato H. Effect of recombinant human platelet-derived growth factor-BB and bone morphogenetic protein-2 application to demineralized dentin on early periodontal ligament cell response. J Periodontal Res 1999;34:244-50. https://doi.org/10.1111/j.1600-0765.1999.tb02250.x
  28. Chen D, Harris MA, Rossini G, Dunstan CR, Dallas SL, Feng JQ, et al. Bone morphogenetic protein 2 (BMP-2) enhances BMP-3, BMP-4, and bone cell differentiation marker gene expression during the induction of mineralized bone matrix formation in cultures of fetal rat calvarial osteoblasts. Calcif Tissue Int 1997;60:283-90. https://doi.org/10.1007/s002239900230
  29. Xu WP, Shiba H, Mizuno N, Uchida Y, Mouri Y, Kawaguchi H, et al. Effect of bone morphogenetic proteins-4, -5 and -6 on DNA synthesis and expression of bone-related proteins in cultured human periodontal ligament cells. Cell Biol Int 2004;28:675-82. https://doi.org/10.1016/j.cellbi.2004.06.004
  30. Nakashima M, Nagasawa H, Yamada Y, Reddi AH. Regulatory role of transforming growth factor-beta, bone morphogenetic protein-2, and protein-4 on gene expression of extracellular matrix proteins and differentiation of dental pulp cells. Dev Biol 1994;162:18-28. https://doi.org/10.1006/dbio.1994.1063
  31. Franceschi RT, Iyer BS. Relationship between collagen synthesis and expression of the osteoblast phenotype in MC3T3-E1 cells. J Bone Miner Res 1992;7:235-46.
  32. Gerstenfeld LC, Chipman SD, Glowacki J, Lian JB. Expression of differentiated function by mineralizing cultures ofchicken osteoblasts. Dev Biol 1987;122:49-60. https://doi.org/10.1016/0012-1606(87)90331-9
  33. Romberg RW, Werness PG, Riggs BL, Mann KG. Inhibition of hydroxyapatite crystal growth by bone-specific and other calcium-binding proteins. Biochemistry 1986;25:1176-80. https://doi.org/10.1021/bi00353a035
  34. Glowacki J, Lian JB. Impaired recruitment and differentiation of osteoclast progenitors by osteocalcin-deplete bone implants. Cell Differ 1987;21:247-54. https://doi.org/10.1016/0045-6039(87)90479-9
  35. Prigodich RV, Vesely MR. Characterization of the complex between bovine osteocalcin and type I collagen. Arch Biochem Biophys 1997;345:339-41. https://doi.org/10.1006/abbi.1997.0254
  36. Ritter NM, Farach-Carson MC, Butler WT. Evidence for the formation of a complex between osteopontin and osteocalcin. J Bone Miner Res 1992;7:877-85.
  37. Aronow MA, Gerstenfeld LC, Owen TA, Tassinari MS, Stein GS, Lian JB. Factors that promote progressive development of the osteoblast phenotype in cultured fetal rat calvaria cells. J Cell Physiol 1990;143:213-21. https://doi.org/10.1002/jcp.1041430203
  38. Malaval L, Modrowski D, Gupta AK, Aubin JE. Cellular expression of bone-related proteins during in vitro osteogenesis in rat bone marrow stromal cell cultures. J Cell Physiol 1994;158:555-72. https://doi.org/10.1002/jcp.1041580322
  39. 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. https://doi.org/10.5051/jkape.2007.37.Suppl.447
  40. Smeyne RJ, Vendrell M, Hayward M, Baker SJ, Miao GG, Schilling K, et al. Continuous c-fos expression precedes programmed cell death in vivo. Nature 1993;363:166-9. https://doi.org/10.1038/363166a0

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