Skip to main content

Advertisement

SpringerLink
Log in

Sonic Hedgehog Promotes Cementoblastic Differentiation via Activating the BMP Pathways

  • Original Research
  • Published:
Calcified Tissue International Aims and scope Submit manuscript

Abstract

Although sonic hedgehog (SHH), an essential molecule in embryogenesis and organogenesis, stimulates proliferation of human periodontal ligament (PDL) stem cells, the effects of recombinant human SHH (rh-SHH) on osteoblastic differentiation are unclear. To reveal the role of SHH in periodontal regeneration, expression of SHH in mouse periodontal tissues and its effects on the osteoblastic/cementoblastic differentiation in human cementoblasts were investigated. SHH is immunolocalized to differentiating cementoblasts, PDL cells, and osteoblasts of the developing mouse periodontium. Addition of rh-SHH increased cell growth, ALP activity, and mineralization nodule formation, and upregulated mRNA expression of osteoblastic and cementoblastic markers. The osteoblastic/cementoblastic differentiation of rh-SHH was abolished by the SHH inhibitor cyclopamine (Cy) and the BMP antagonist noggin. rh-SHH increased the expression of BMP-2 and -4 mRNA, as well as levels of phosphorylated Akt, ERK, p38, and JNK, and of MAPK and NF-κB activation, which were reversed by noggin, Cy, and BMP-2 siRNA. Collectively, this study is the first to demonstrate that SHH can promote cell growth and cell osteoblastic/cementoblastic differentiation via BMP pathway. Thus, SHH plays important roles in the development of periodontal tissue, and might represent a new therapeutic target for periodontitis and periodontal regeneration.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7

Similar content being viewed by others

References

  1. Bartold PM, McCulloch CA, Narayanan AS, Pitaru S (2000) Tissue engineering: a new paradigm for periodontal regeneration based on molecular and cell biology. Periodontology 24:253–269

    Article  CAS  Google Scholar 

  2. Wang HL, Greenwell H, Fiorellini J, Giannobile W, Offenbacher S, Salkin L, Townsend C, Sheridan P, Genco RJ (2005) Periodontal regeneration. J Periodontol 76(9):1601–1622

    Article  CAS  PubMed  Google Scholar 

  3. Menicanin D, Hynes K, Han J, Gronthos S, Bartold PM (2015) Cementum and periodontal ligament regeneration. Adv Exp Med Biol 881:207–236

    Article  PubMed  Google Scholar 

  4. Foster BL, Somerman MJ (2012) Cementum. In: McCauley LK, Somerman MJ (eds) Mineralized tissues in oral and craniofacial science: biological principles and clinical correlates, 1st edn. Wiley-Blackwell, Ames, pp 169–192

    Google Scholar 

  5. Kitagawa M, Tahara H, Kitagawa S, Oka H, Kudo Y, Sato S, Ogawa I, Miyaichi M, Takata T (2006) Characterization of established cementoblast-like cell lines from human cementum-lining cells in vitro and in vivo. Bone 39(5):1035–1042

    Article  CAS  PubMed  Google Scholar 

  6. Lee SI, Lee DW, Yun HM, Cha HJ, Bae CH, Cho ES, Kim EC (2015) Expression of thymosin beta-4 in human periodontal ligament cells and mouse periodontal tissue and its role in osteoblastic/cementoblastic differentiation. Differentiation 90(1–3):16–26

    Article  CAS  PubMed  Google Scholar 

  7. Jin H, Choung HW, Lim KT, Jin B, Jin C, Chung JH, Choung PH (2015) Recombinant human plasminogen activator inhibitor-1 promotes cementogenic differentiation of human periodontal ligament stem cells. Tissue Eng Part A 21(23–24):2817–2828

    Article  CAS  PubMed  Google Scholar 

  8. Hoz L, Romo E, Zeichner-David M, Sanz M, Nuñez J, Gaitán L, Mercado G, Arzate H (2012) Cementum protein 1 (CEMP1) induces differentiation by human periodontal ligament cells under three-dimensional culture conditions. Cell Biol Int 36(2):129–136

    Article  CAS  PubMed  Google Scholar 

  9. Bosshardt DD, Sculean A, Windisch P, Pjetursson BE, Lang NP (2005) Effects of enamel matrix proteins on tissue formation along the roots of human teeth. J Periodontal Res 40(2):158–167

    Article  CAS  PubMed  Google Scholar 

  10. Lee SY, Auh QS, Kang SK, Kim HJ, Lee JW, Noh K, Jang JH, Kim EC (2014) Combined effects of dentin sialoprotein and bone morphogenetic protein-2 on differentiation in human cementoblasts. Cell Tissue Res 357(1):119–132

    Article  CAS  PubMed  Google Scholar 

  11. Ingham PW, McMahon AP (2001) Hedgehog signaling in animal development: paradigms and principles. Genes Dev 15(23):3059–3087

    Article  CAS  PubMed  Google Scholar 

  12. Choudhry Z, Rikani AA, Choudhry AM, Tariq S, Zakaria F, Asghar MW, Sarfraz NK, Haider K, Shafiq AA, Mobassarah NJ (2014) Sonic hedgehog signaling pathway: a complex network. Ann Neurosci 21(1):28–31

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  13. Hu X, Huang J, Feng L, Fukudome S, Hamajima Y, Lin J (2010) Sonic hedgehog (SHH) promotes the differentiation of mouse cochlear neural progenitors via the Math1-Brn3.1 signaling pathway in vitro. J Neurosci Res 88(5):927–935

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  14. Elia D, Madhala D, Ardon E, Reshef R, Halvey O (2007) Sonic hedgehog promotes proliferation and differentiation of adult muscle cells: involvement of MAPK/ERK and PI3K/Akt pathways. Biochim Biophys Acta 1773(9):1438–1446

    Article  CAS  PubMed  Google Scholar 

  15. Dutton R, Yamada T, Turnley A, Bartlett PF, Murphy M (1999) Sonic hedgehog promotes neuronal differentiation of murine spinal cord precursors and collaborates with neurotrophin 3 to induce Islet-1. J Neurosci 19(7):2601–2608

    CAS  PubMed  Google Scholar 

  16. Warzecha J, Göttig S, Brüning C, Lindhorst E, Arabmothlagh M, Kuth A (2006) Sonic hedgehog protein promotes proliferation and chondrogenic differentiation of bone marrow-derived mesenchymal stem cells in vitro. J Orthop Sci 11(5):491–496

    Article  CAS  PubMed  Google Scholar 

  17. Chiang C, Litingtung Y, Lee E, Young KE, Corden JL, Westphal H, Beachy PA (1996) Cyclopia and defective axial patterning in mice lacking sonic hedgehog gene function. Nature 383(6599):407–413

    Article  CAS  PubMed  Google Scholar 

  18. Miyaji T, Nakase T, Iwasaki M, Kuriyama K, Tamai N, Highchi C, Myoui A, Tomita T, Yoshigawa H (2003) Expression and distribution of transcripts for sonic hedgehog in the early phase of fracture repair. Histochem Cell Biol 119(3):233–237

    CAS  PubMed  Google Scholar 

  19. Levi B, James AW, Nelson ER, Li S, Peng M, Commons GW, Lee M, Wu B, Longaker MT (2011) Human adipose-derived stromal cells stimulate autogenous skeletal repair via paracrine Hedgehog signaling with calvarial osteoblasts. Stem Cells Dev 20(2):243–257

    Article  CAS  PubMed  Google Scholar 

  20. Gritli-Linde A, Bei M, Maas R, Zhang XM, Linde A, McMahon AP (2002) Shh signaling within the dental epithelium is necessary for cell proliferation, growth and polarization. Development 129(23):5323–5337

    Article  CAS  PubMed  Google Scholar 

  21. Seidel K, Ahn CP, Lyons D, Nee A, Ting K, Brwonell I, Cao T, Carano RA, Curran T, Schober M, Fuchs E, Joyner A, Marin GR, de Sauvage FJ, Klein OD (2010) Hedgehog signaling regulates the generation of ameloblast progenitors in the continuously growing mouse incisor. Development 137(22):3753–3761

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  22. Gaspard N, Bouschet T, Hourez R, Dimidschstein J, Naeije G, van den Ameele J, Espuny-Camacho I, Herpoel A, Passante L, Scjiffmann SN, Gaillard A, Vanderhaeghen P (2008) An intrinsic mechanism of corticogenesis from embryonic stem cells. Nature 455(7211):351–357

    Article  CAS  PubMed  Google Scholar 

  23. James AW, Leucht P, Levi B, Carre AL, Xu Y, Helms JA, Longaker MT (2010) Sonic Hedgehog influences the balance of osteogenesis and adipogenesis in mouse adipose-derived stromal cells. Tissue Eng Part A 16(8):2605–2616

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  24. Spinella-Jaegle S, Rawadi G, Kawai S, Gallea S, Faucheu C, Mollat P, Courtois B, Bergaud B, Ramez V, Blanchet AM, Adelmant G, Baron R, Roman-Roman S (2001) Sonic hedgehog increases the commitment of pluripotent mesenchymal cells into the osteoblastic lineage and abolishes adipocytic differentiation. J Cell Sci 114(Pt 11):2085–2094

    CAS  PubMed  Google Scholar 

  25. Tian Y, Xu Y, Fu Q, Dong Y (2012) Osterix is required for Sonic hedgehog-induced osteoblastic MC3T3-E1 cell differentiation. Cell Biochem Biophys 64(3):169–176

    Article  CAS  PubMed  Google Scholar 

  26. Ho JE, Chung EH, Wall S, Schaffer DV, Healy KE (2007) Immobilized sonic hedgehog N-terminal signaling domain enhances differentiation of bone marrow-derived mesenchymal stem cells. J Biomed Mater Res A 83(4):1200–1208

    Article  PubMed  Google Scholar 

  27. van der Horst G, Farih-Sips H, Löwik CW, Karperien M (2003) Hedgehog stimulates only osteoblastic differentiation of undifferentiated KS483 cells. Bone 33(6):899–910

    Article  PubMed  Google Scholar 

  28. Jemtland R, Divieti P, Lee K, Segre GV (2003) Hedgehog promotes primary osteoblast differentiation and increases PTHrP mRNA expression and iPTHrP secretion. Bone 32(6):611–620

    Article  CAS  PubMed  Google Scholar 

  29. Xia L, Zhang M, Chang Q, Wang L, Zeng D, Zhang X, Zhang Z, Jiang X (2013) Enhanced dentin-like mineralized tissue formation by AdShh-transfected human dental pulp cells and porous calcium phosphate cement. PLoS One 8(5):e62645

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  30. Takahashi S, Kawashima N, Sakamoto K, Nakata A, Kameda T, Sugiyama AT, Katsube K, Suda H (2007) Differentiation of an ameloblast-lineage cell line (ALC) is induced by Sonic hedgehog signaling. Biochem Biophys Res Commun 353(2):405–411

    Article  CAS  PubMed  Google Scholar 

  31. Jiang Q, Du J, Yin X, Shan Z, Ma Y, Ma P, Du J, Fan Z (2013) Shh signaling, negatively regulated by BMP signaling, inhibits the osteo/dentinogenic differentiation potentials of mesenchymal stem cells from apical papilla. Mol Cell Biochem 383(1–2):85–93

    Article  CAS  PubMed  Google Scholar 

  32. Plaisant M, Fontaine C, Cousin W, Rochet N, Dani C, Peraldi P (2009) Activation of hedgehog signaling inhibits osteoblast differentiation of human mesenchymal stem cells. Stem Cells 27(3):703–713

    Article  CAS  PubMed  Google Scholar 

  33. Krishnan V, Ma Y, Moseley J, Geiser A, Fraint S, Frolik C (2001) Bone anabolic effects of Sonic/Indian hedgehog are mediated by BMP-2/4-dependent pathways in the neonatal rat metatarsal model. Endocrinology 142(2):940–947

    Article  CAS  PubMed  Google Scholar 

  34. Spinella-Jaeqle S, Rawadi G, Kawai S, Gallea S, Faucheu C, Mollat P (2001) Sonic hedgehog increases the commitment of pluripotent mesenchymal cells into the osteoblastic lineage and abolishes adipocytic differentiation. J Cell Sci 114(Pt 111):2085–2094

    Google Scholar 

  35. Hammerschmidt M, Brook A, McMahon AJ (1997) The world according to hedgehog. Trends Genet 13(1):14–21

    Article  CAS  PubMed  Google Scholar 

  36. Bitgood MJ, McMahon AP (1995) Hedgehog and Bmp genes are coexpressed at many diverse sites of cell-cell interaction in the mouse embryo. Dev Biol 172(1):126–138

    Article  CAS  PubMed  Google Scholar 

  37. Martínez C, Smith PC, Rodriguez JP, Palma V (2011) Sonic hedgehog stimulates proliferation of human periodontal ligament stem cells. J Dent Res 90(4):483–488

    Article  PubMed  Google Scholar 

  38. Dassule HR, Lewis P, Bei M, Maas R, McMahon AP (2000) Sonic hedgehog regulates growth and morphogenesis of the tooth. Development 127(22):4775–4785

    CAS  PubMed  Google Scholar 

  39. Wu C, Shimo T, Liu M, Pacifici M, Koyama E (2003) Sonic hedgehog functions as a mitogen during bell stage of odontogenesis. Connect Tissue Res 44(Suppl 1):92–96

    Article  CAS  PubMed  Google Scholar 

  40. Gulino R, Gulisano M (2013) Noggin and Sonic hedgehog are involved in compensatory changes within the motoneuron-depleted mouse spinal cord. J Neurol Sci 332(1–2):102–109

    Article  CAS  PubMed  Google Scholar 

  41. van den Brink GR, Hardwick JC, Nielsen C, Xu C, ten Kate FJ, Glickman J, van Deventer SJ, Roberts DJ, Peppelenbosch MP (2002) Sonic hedgehog expression correlates with fundic gland differentiation in the adult gastrointestinal tract. Gut 51(5):628–633

    Article  PubMed  PubMed Central  Google Scholar 

  42. Lai K, Kaspar BK, Gage FH, Schaffer DV (2003) Sonic hedgehog regulates adult neural progenitor proliferation in vitro and in vivo. Nat Neurosci 6(1):21–27

    Article  CAS  PubMed  Google Scholar 

  43. Khan M, Seppala M, Zoupa M, Cobourne MT (2007) Hedgehog pathway gene expression during early development of the molar tooth root in the mouse. Gene Exp Patterns 7(3):239–243

    Article  CAS  Google Scholar 

  44. Ming JE, Roessler E, Muenke M (1998) Human developmental disorders and the Sonic hedgehog pathway. Mol Med Today 4(8):343–349

    Article  CAS  PubMed  Google Scholar 

  45. Nakatomi M, Morita I, Eto K, Ota MS (2006) Sonic hedgehog signaling is important in tooth root development. J Dent Res 85(5):427–431

    Article  CAS  PubMed  Google Scholar 

  46. Nakatomi M, Hovorakova M, Gritli-Linde A, Blair HJ, MacArthur K, Peterka M, Lesot H, Peterkova R, Ruiz-Perez VL, Goodship JA, Peters H (2013) Evc regulates a symmetrical response to Shh signaling in molar development. J Dent Res 92(3):222–228

    Article  CAS  PubMed  Google Scholar 

  47. Alvarez-Pérez MA, Narayanan S, Zeichner-David M, Rodríguez Carmona B, Arzate H (2006) Molecular cloning, expression and immunolocalization of a novel human cementum-derived protein (CP-23). Bone 38(3):409–419

    Article  PubMed  Google Scholar 

  48. Arzate H, Olson SW, Page RC, Narayanan AS (1992) Isolation of human tumor cells that produce cementum proteins in culture. Bone Miner 18(1):15–30

    Article  CAS  PubMed  Google Scholar 

  49. Saito M, Iwase M, Maslan S, Nozaki N, Yamauchi M, Handa K, Takahashi O, Sato S, Kawase T, Teranaka T, Narayanan AS (2001) Expression of cementum-derived attachment protein in bovine tooth germ during cementogenesis. Bone 29(3):242–248

    Article  CAS  PubMed  Google Scholar 

  50. Eipeldauer S, Thomas A, Hoechtl-Lee L, Kecht M, Binder H, Koettstorfer J, Gregori M, Sarahrudi K (2014) Is sonic Hedgehog involved in human fracture healing?—a prospective study on local and systemic concentrations of SHH. PLoS One 9(12):668

    Article  Google Scholar 

  51. Cai JQ, Huang YZ, Chen XH, Xie HL, Zhu HM, Tang L, Yang ZM, Huang YC, Deng L (2012) Sonic hedgehog enhances the proliferation and osteogenic differentiation of bone marrow-derived mesenchymal stem cells. Cell Biol Int 36(4):349–355

    Article  CAS  PubMed  Google Scholar 

  52. Fu JR, Liu WL, Zhou JF, Sun HY, Xu HZ, Luo L, Zhang H, Zhou YF (2006) Sonic hedgehog protein promotes bone marrow-derived endothelial progenitor cell proliferation, migration and VEGF production via PI 3-kinase/Akt signaling pathways. Acta Pharmacol Sin 27(6):685–693

    Article  CAS  PubMed  Google Scholar 

  53. Luo JD, Hu TP, Wang L, Chen MS, Liu SM, Chen AF (2009) Sonic hedgehog improves delayed wound healing via enhancing cutaneous nitric oxide function in diabetes. Am J Physiol Endocrinol Metab 297(2):E525–E531

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  54. Asai J, Takenaka H, Kusano KF, Ii M, Luedemann C, Curry C, Eaton E, Iwakura A, Tsutsumi Y, Hamada H, Kishimoto S, Thorne T, Kishore R, Losordo DW (2006) Topical sonic hedgehog gene therapy accelerates wound healing in diabetes by enhancing endothelial progenitor cell-mediated microvascular remodeling. Circulation 113(20):2413–2424

    Article  CAS  PubMed  Google Scholar 

  55. Edwards PC, Ruggiero S, Fantasia J, Burakoff R, Moorji SM, Paric E, Razzano P, Grande DA, Mason JM (2005) Sonic hedgehog gene-enhanced tissue engineering for bone regeneration. Gene Ther 12(1):75–86

    Article  CAS  PubMed  Google Scholar 

  56. Chen JK, Taipale J, Cooper MK, Beachy PA (2002) Inhibition of Hedgehog signaling by direct binding of cyclopamine to Smoothened. Genes Dev 16(21):2743–2748

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  57. Yun HM, Park KR, Quang TH, Oh H, Hong JT, Kim YC, Kim EC (2015) 2,4,5-Trimethoxyldalbergiquinol promotes osteoblastic differentiation and mineralization via the BMP and Wnt/β-catenin pathway. Cell Death Dis 6:e1819

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  58. Hallahan AR, Pritchard JI, Chandraratna RA, Ellenbogen RG, Geyer JR, Overland RP, Strand AD, Tapscott SJ, Olson JM (2003) BMP-2 mediates retinoid-induced apoptosis in medulloblastoma cells through a paracrine effect. Nat Med 9(8):1033–1038

    Article  CAS  PubMed  Google Scholar 

  59. Gazzerro E, Gangji V, Canalis E (1998) Bone morphogenetic proteins induce the expression of noggin, which limits their activity in cultured rat osteoblasts. J Clin Invest 102(12):2106–2114

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  60. Devlin RD, Du Z, Pereira RC, Kimble RB, Economides AN, Jorgetti V, Canalis E (2003) Skeletal overexpression of noggin results in osteopenia and reduced bone formation. Endocrinology 144(5):1972–1978

    Article  CAS  PubMed  Google Scholar 

  61. Xiao G, Gopalakrishnan R, Jiang D, Reith E, Benson M, Franceschi RT (2002) Bone morphogenetic proteins, extracellular matrix, and mitogen-activated protein kinase signaling pathways are required for osteoblast-specific gene expression and differentiation in MC3T3-E1 cells. J Bone Miner Res 17(1):101–110

    Article  CAS  PubMed  Google Scholar 

  62. Ghosh-Choudhury N, Mandal CC, Choudhury GG (2007) Statin-induced Ras activation integrates the phosphatidylinositol 3-kinase signal to Akt and MAPK for bone morphogenetic protein-2 expression in osteoblast differentiation. J Biol Chem 282(7):4983–4993

    Article  CAS  PubMed  Google Scholar 

  63. Dormoy V, Danilin S, Lindner V, Thomas L, Rothhut S, Coquard C, Helwig JJ, Jacqmin D, Lang H, Massfelder T (2009) The sonic hedgehog signaling pathway is reactivated in human renal cell carcinoma and plays orchestral role in tumor growth. Mol Cancer 16(8):123

    Article  Google Scholar 

Download references

Acknowledgments

This work was supported by a National Research Foundation of Korea (NRF) grant funded by the Korean government (MEST) (No. 2012R1A5A2051384).

Author information

Author notes

    Authors and Affiliations

    1. Department of Oral and Maxillofacial Pathology, School of Dentistry and Research Center for Tooth & Periodontal Regeneration (MRC), Kyung Hee University, 14 Kyungheedae-ro, Dongdaemun-gu, Seoul, 02453, Republic of Korea

      Won-Jung Bae & Eun-Cheol Kim

    2. Department of Oral Medicine, School of Dentistry, Kyung Hee University, Seoul, Republic of Korea

      Q-Schick Auh

    3. Department of Periodontology, School of Dentistry, Kyung Hee University, Seoul, Republic of Korea

      Hyun-Chang Lim

    4. Department of Oral and Maxillofacial Radiology, School of Dentistry, Kyung Hee University, Seoul, Republic of Korea

      Gyu-Tae Kim

    5. Department of Orthodontics, Graduate School, Kyung Hee University, Seoul, Republic of Korea

      Hyun-Soo Kim

    Authors
    1. Won-Jung Bae
      View author publications

      You can also search for this author in PubMed Google Scholar

    2. Q-Schick Auh
      View author publications

      You can also search for this author in PubMed Google Scholar

    3. Hyun-Chang Lim
      View author publications

      You can also search for this author in PubMed Google Scholar

    4. Gyu-Tae Kim
      View author publications

      You can also search for this author in PubMed Google Scholar

    5. Hyun-Soo Kim
      View author publications

      You can also search for this author in PubMed Google Scholar

    6. Eun-Cheol Kim
      View author publications

      You can also search for this author in PubMed Google Scholar

    Corresponding author

    Correspondence to Eun-Cheol Kim.

    Ethics declarations

    Conflict of Interest

    Won-Jung Bae, Q-Schick Auh, Hyun-Chang Lim, Gyu-Tae Kim, Hyun-Soo Kim and Eun-Cheol Kim declare that they have no conflict of interest.

    Human and Animal Rights and Informed Consent

    The animal experimental procedures were performed in accordance with the ethical guidelines and were approved by the Kyung Hee University Animal Care Committee (Seoul, Korea).

    Additional information

    Won-Jung Bae and Q-Schick Auh have contributed equally to this manuscript.

    Rights and permissions

    Reprints and permissions

    About this article

    Check for updates. Verify currency and authenticity via CrossMark

    Cite this article

    Bae, WJ., Auh, QS., Lim, HC. et al. Sonic Hedgehog Promotes Cementoblastic Differentiation via Activating the BMP Pathways. Calcif Tissue Int 99, 396–407 (2016). https://doi.org/10.1007/s00223-016-0155-1

    Download citation

    • Received:

    • Accepted:

    • Published:

    • Issue Date:

    • DOI: https://doi.org/10.1007/s00223-016-0155-1

    Keywords

    Search

    Navigation