International Journal of Oral Biology

  • 제30권3호
  • /
  • Pages.99-103
  • /
  • 2005
  • /
  • 1226-7155(pISSN)
  • /
  • 2287-6618(eISSN)
대한구강생물학회 (The Korean Academy of Oral Biology)
권호 표지

BMP Expression by Human Cementum-Derived Cells in vitro

  • Ko, Hyun-Jung (Department of Anatomy and Cell Biology, School of Dental Medicine, University of Pennsylvania) ;
  • Grzesik, Wojciech J (Department of Anatomy and Cell Biology, School of Dental Medicine, University of Pennsylvania)
  • 발행 : 2005.09.30
PDF 다운로드
⟨ 이전 논문 다음 논문 ⟩

초록

Bone morphogenetic proteins (BMPs), members of a large group of TGF-beta family, are important molecular regulators of morphogenesis of numerous tissues and organs, including bones and teeth. Most BMPs are capable of inducing bone formation in vivo and therefore are of considerable clinical interest for regenerating mineralized tissues. Recently, we have developed a method to culture cells from human cementum (human cementum-derived cells, HCDCs). HCDCs, when attached to synthetic hydroxyapatite/tricalcium phosphate (HA/TCP) ceramic and transplanted into immunodeficient mice, formed histologically identifiable cementum-like tissue. Since it is unclear to what extent BMPs are involved in cementogenesis, the aim of this study was to establish which BMPs are expressed by cementogenic HCDCs and whether the expression of BMPs is related to the degree of cellular differentiation in vitro. HCDCs were maintained in growth medium (DMEM/F12 supplemented with 10% FBS) until confluent (proliferation stage). Upon reaching confluence, cells were incubated in the differentiation medium (DMEM/F12 medium containing 10% FBS and 50 mg/ml ascorbic acid) for 14 days (differentiation stage). Next, HCDCs were incubated in mineralization medium (DMEM/F12, 50 mg/ml ascorbic acid, 2.5 mg/ml of ITS (insulin-transferrinselenium), 5 mM beta-glycerophosphate and $10^{-8}M$ dexamethasone) for another 14 days (mineralization stage). At the end of each differentiation stage, total RNA was isolated and evaluated for BMPs (2 through 8) expression by employing real time RT-PCR. HCDCs expressed most of BMPs examined except BMP-7 and BMP-8. Furthermore, on average, the highest levels of BMPs were expressed at the earlier differentiation stage, prior to the initiation of mineralization in vitro. These results indicate that several BMPs are expressed during cementoblastic differentiation and suggest that BMPs may be involved in the homeostasis of human cementum.

키워드

참고문헌

  1. Aberg, T., Wozney, J., Thesleff, I.: Expression patterns of bone morphogenetic proteins (Bmps) in the developing mouse tooth suggest roles in morphogenesis and cell differentiation. Dev. Dyn. 210:383-396, 1997 https://doi.org/10.1002/(SICI)1097-0177(199712)210:4<383::AID-AJA3>3.0.CO;2-C
  2. Alatli, I., Lundmark, C., Hammarstrom, L.: The localization of epithelial root sheath cells during cementum formation in rat molars. J. Periodontal. Res. 31:433-440, 1996 https://doi.org/10.1111/j.1600-0765.1996.tb00512.x
  3. Bianco, P., Kuznetsov, S.A., Riminucci, M., Fisher, L.W., Spiegel, A.M., Robey, P.G.: A miniature of human fibrous dysplasia of bone reproduced in immunodeficient mice by transplantation of mosaics of normal and Gs alpha-mutated marrow stromal fibroblasts. J. Clin. Invest. 101:1737-1744, 1998 https://doi.org/10.1172/JCI2361
  4. Chen, A. L., Fang, C., Liu, C., Leslie, M.P., Chang, E., Di Cesare, P.E.: Expression of bone morphogenetic proteins, receptors, and tissue inhibitors in human fetal, adult, and osteoarthritic articular cartilage. J. Orthopaedic. Res. 22: 1188-1192, 2004 https://doi.org/10.1016/j.orthres.2004.02.013
  5. Daluiski, A., Engstrand, T., Bahamonde, M.E., Gamer, L.W., Agius, E., Stevenson, S.L., Cox, K., Rosen, V., Lyons, K.M.: Bone morphogenetic protein-3 is a negative regulator of bone density. Nature Genetics. 27:84-88, 2001 https://doi.org/10.1038/83810
  6. Dudas, M., Sridurongrit, S., Nagy, A., Okazaki, K., Kaartinen, V.: Craniofacial defects in mice lacking BMP type I receptor Alk2 in neural crest cells. Mech. Dev. 121:173-182, 2004 https://doi.org/10.1016/j.mod.2003.12.003
  7. Freeman, E.: Periodontium. Oral Histology, 4th Ed. Mosby-Year Book, Inc., St. Louis, MO, U.S.A., pp. 276-312, 1994
  8. Grant, D.A., Stern, I.B., Listgarten, M.A.: Cementum. Periodontics, 6th Ed. The CV Mosby Company, St. Louis, MO, U.S.A., pp. 76-93, 1988
  9. Grzesik, W.J., Kuznetsov, S.A., Uzawa, K., Mankani, M., Robey, P.G., Yamauchi, M.: Normal human cementum-derived cells: isolation, clonal expansion and in vitro and in vivo characterization. J. Bone. Miner. Res. 13:1547-1554, 1998 https://doi.org/10.1359/jbmr.1998.13.10.1547
  10. Hogan, B.L.: Bone morphogenetic proteins: Multifunctional regulators of vertebrate development. Genes. Dev. 10:1580-1594, 1996 https://doi.org/10.1101/gad.10.13.1580
  11. Kingsley, D.M.: The TGF-beta superfamily: New members, new receptors, and new genetic tests of function in different organisms. Genes. Dev. 8:133-146, 1994 https://doi.org/10.1101/gad.8.2.133
  12. Lu, H., Jin, Y., Tipoe, G.L.: Alteration in the expression of bone morphogenetic protein-2,3,4,5 mRNA during pathogenesis of cleft palate in BALB/c mice. Arch. Oral. Biol. 45:133-140, 2000 https://doi.org/10.1016/S0003-9969(99)00118-1
  13. Luyten, F.P., Cunningham, N.S., Ma, S., Muthukumaran, N., Hammonds, R.G., Nevins, W.B., Woods, W.I., Reddi, A.H.: Purification and partial amino acid sequence of osteogenin, a protein initiating bone differentiation. J. Biol. Chem. 264: 13377-13380, 1989
  14. Reddi, A.H.: Bone morphogenetic proteins: An unconventional approach to isolation of first mammalian morphogens. Cytokine. Growth. Factor. Rev. 8:11 -20, 1997 https://doi.org/10.1016/S1359-6101(96)00049-4
  15. Takao, M., Hino, J., Takeshita, N., Konno, Y, Nishizawa, T., Matsuo, H., Kangawa, K.: Identification of rat bone morphogenetic protein-3b (BMP3b), a new member of BMP-3. Biochem. Biophys. Res. Commun. 219:656-662, 1996 https://doi.org/10.1006/bbrc.1996.0289
  16. Thesleff, I.: Homeobox genes and growth factors in regulation of craniofacial and tooth morphogenesis. Acta. Odontol. Scand. 53:129-134, 1995 https://doi.org/10.3109/00016359509005962
  17. Thomadakis, G., Ramoshebi, L.N., Crooks, J., Rueger, D.C., Ripamonti, U.: Immunolocalization of Bone Morphogenetic Protein-2 and -3 and Osteogenic Protein-1 during murine tooth root morphogenesis and in other craniofacial structures. Eur. J. Oral. Sci. 107:368-377, 1999 https://doi.org/10.1046/j.0909-8836.1999.eos107508.x
  18. Urist, M.R.: Bone: formation by autoinduction. Science. 150: 893-899, 1965 https://doi.org/10.1126/science.150.3698.893
  19. Wang, E.A., V. Rosen, J.S., D'Alessandro, M., Bauduy, M., Cordes, P., Harada, T., Israel, D.I., Hewick, R.M., Kerns, K.M., LaPan, P., et al. Recombinant human bone morphogenetic protein induces bone formation. Proc. Natl. Acad. Sci. USA. 87: 2220-2224, 1990 https://doi.org/10.1073/pnas.87.6.2220
  20. Wozney, J.M., Rosen, V., Celeste, A.J., Mitsock, L.M., Whitters, M.J., Kriz, R.W., Hewick, R.M., Wang, E.A.: Novel regulators of bone formation: Molecular clones and activities. Science. 242:1528-1534, 1988 https://doi.org/10.1126/science.3201241
  21. Wozney, J.M. & Rosen, V.: in Physiology and Pharmacology of Bone pp.725-748, Springer-Verlag, Berlin, Germany, 1993
  22. Yamamoto, T., Hinrichsen, K.V.: The development of cellular cementum in rat molars, with special reference to the fiber arrangement. Anat. Embryol. 188:537-549, 1993 https://doi.org/10.1007/BF00187009
  23. Yamashiro, T., Tummers, M Thesleff, I.: Expression of Bone Morphogenetic Proteins and Msx Genes during Root Formation. J. Dent. Res. 82:172-176, 2003 https://doi.org/10.1177/154405910308200305
  24. Yamashita, H., Ten Dijke, P., Heldin, C.H., Miyazono, K.: Bone morphogenetic protein receptors. Bone. 19:569-574, 1996 https://doi.org/10.1016/S8756-3282(96)00259-1