Ligand–Receptor Interactions and Their Implications in Delivering Certain Signaling for Bone Regeneration

  • Takenobu KatagiriEmail author
  • Sho Tsukamoto
  • Kenji Osawa
  • Shoichiro Kokabu
Part of the Mechanical Engineering Series book series (MES)


Cartilage and bone tissue formation is observed not only during embryonic development but also in some pathological conditions occurring after birth, including fracture healing. This process is regulated by many stimuli that are applicable to the reconstitution of skeletal tissues using tissue engineering. In particular, members of the transforming growth factor (TGF)-β family play a unique and important role in skeletal tissue formation, wherein they activate specific intracellular signaling pathways by binding two types of serine–threonine kinase receptors and downstream effectors called Smad proteins. The biological activity of TGF-β family members is positively and negatively regulated at multiple steps by various molecules found in the extracellular space, on the cell membrane, and in the intracellular space. The modification of TGF-β family signaling pathways can be used in tissue engineering approaches for skeletal tissue formation.


Bone Morphogenetic Protein Osteoblastic Differentiation C2C12 Cell Demineralized Bone Matrix Ectopic Bone Formation 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.



We thank the members of the Division of Pathophysiology, Research Center for Genomic Medicine, Saitama Medical University for their valuable comments and discussion. This work was supported in part by Grants-in-Aid from the Ministry of Education, Culture, Sports, Science, and Technology of Japan and a Grant-in-Aid from the Support Project for the Formation of a Strategic Center in a Private University from the Ministry of Education, Culture, Sports, Science, and Technology of Japan.


  1. Aono A, Hazama M, Notoya K, Taketomi S, Yamasaki H, Tsukuda R, Sasaki S, Fujisawa Y (1995) Potent ectopic bone-inducing activity of bone morphogenetic protein-4/7 heterodimer. Biochem Biophys Res Commun 210:670–677CrossRefGoogle Scholar
  2. Balemans W et al (2001) Increased bone density in sclerosteosis is due to the deficiency of a novel secreted protein (SOST). Hum Mol Genet 10:537–543CrossRefGoogle Scholar
  3. Baron R, Kneissel M (2013) WNT signaling in bone homeostasis and disease: from human mutations to treatments. Nat Med 19:179–192CrossRefGoogle Scholar
  4. Bessa PC, Cerqueira MT, Rada T, Gomes ME, Neves NM, Nobre A, Reis RL, Casal M (2009) Expression, purification and osteogenic bioactivity of recombinant human BMP-4, -9, -10, -11 and -14. Protein Expr Purif 63:89–94CrossRefGoogle Scholar
  5. Blau HM, Chiu CP, Webster C (1983) Cytoplasmic activation of human nuclear genes in stable heterocaryons. Cell 32:1171–1180CrossRefGoogle Scholar
  6. Boyden LM et al (2002) High bone density due to a mutation in LDL-receptor-related protein 5. N Engl J Med 346:1513–1521CrossRefGoogle Scholar
  7. Bruce DL, Sapkota GP (2012) Phosphatases in SMAD regulation. FEBS Lett 586:1897–1905CrossRefGoogle Scholar
  8. Brunet LJ, McMahon JA, McMahon AP, Harland RM (1998) Noggin, cartilage morphogenesis, and joint formation in the mammalian skeleton. Science 280:7–1455CrossRefGoogle Scholar
  9. Brunkow ME et al (2001) Bone dysplasia sclerosteosis results from loss of the SOST gene product, a novel cystine knot-containing protein. Am J Hum Genet 68:89–577CrossRefGoogle Scholar
  10. Celeste AJ, Iannazzi JA, Taylor RC, Hewick RM, Rosen V, Wang EA, Wozney JM (1990) Identification of transforming growth factor beta family members present in bone-inductive protein purified from bovine bone. Proc Natl Acad Sci USA 87:9843–9847CrossRefGoogle Scholar
  11. Daluiski A et al (2001) Bone morphogenetic protein-3 is a negative regulator of bone density. Nat Genet 27:84–88Google Scholar
  12. Ellies DL, Viviano B, McCarthy J, Rey JP, Itasaki N, Saunders S, Krumlauf R (2006) Bone density ligand, Sclerostin, directly interacts with LRP5 but not LRP5G171 V to modulate Wnt activity. J Bone Miner Res 21:1738–1749CrossRefGoogle Scholar
  13. Fukuda T et al (2010) Canonical Wnts and BMPs cooperatively induce osteoblastic differentiation through a GSK3beta-dependent and beta-catenin-independent mechanism. Differentiation 80:46–52CrossRefGoogle Scholar
  14. 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 102:2106–2114CrossRefGoogle Scholar
  15. Gong Y et al (1999) Heterozygous mutations in the gene encoding noggin affect human joint morphogenesis. Nat Genet 21:302–304CrossRefGoogle Scholar
  16. Gong Y et al (2001) LDL receptor-related protein 5 (LRP5) affects bone accrual and eye development. Cell 107:513–523CrossRefGoogle Scholar
  17. Groppe J et al (2002) Structural basis of BMP signalling inhibition by the cystine knot protein Noggin. Nature 420:636–642CrossRefGoogle Scholar
  18. Groppe J et al (2008) Cooperative assembly of TGF-beta superfamily signaling complexes is mediated by two disparate mechanisms and distinct modes of receptor binding. Mol Cell 29:157–168CrossRefGoogle Scholar
  19. Huse M, Muir TW, Xu L, Chen YG, Kuriyan J, Massague J (2001) The TGF beta receptor activation process: an inhibitor-to substrate-binding switch. Mol Cell 8:671–682CrossRefGoogle Scholar
  20. Israel DI, Nove J, Kerns KM, Kaufman RJ, Rosen V, Cox KA, Wozney JM (1996) Heterodimeric bone morphogenetic proteins show enhanced activity in vitro and in vivo. Growth Factors 13:291–300CrossRefGoogle Scholar
  21. Kang Q et al (2004) Characterization of the distinct orthotopic bone-forming activity of 14 BMPs using recombinant adenovirus-mediated gene delivery. Gene Ther 11:1312–1320CrossRefGoogle Scholar
  22. Kanomata K, Kokabu S, Nojima J, Fukuda T, Katagiri T (2009) DRAGON, a GPI-anchored membrane protein, inhibits BMP signaling in C2C12 myoblasts. Genes Cells 14:695–702CrossRefGoogle Scholar
  23. Katagiri T (2010) Heterotopic bone formation induced by bone morphogenetic protein signaling: fibrodysplasia ossificans progressiva. J Oral Biosci 52:33–41CrossRefGoogle Scholar
  24. Katagiri T, Tsukamoto S (2013) The unique activity of bone morphogenetic proteins in bone: a critical role of the Smad signaling pathway. Biol Chem 394:703–714CrossRefGoogle Scholar
  25. Katagiri T et al (1994) Bone morphogenetic protein-2 converts the differentiation pathway of C2C12 myoblasts into the osteoblast lineage. J Cell Biol 127:1755–1766CrossRefGoogle Scholar
  26. Kato Y, Iwamoto M, Koike T, Suzuki F, Takano Y (1988) Terminal differentiation and calcification in rabbit chondrocyte cultures grown in centrifuge tubes: regulation by transforming growth factor beta and serum factors. Proc Natl Acad Sci USA 85:9552–9556CrossRefGoogle Scholar
  27. Kokabu S et al (2012a) BMP3 suppresses osteoblast differentiation of bone marrow stromal cells via interaction with Acvr2b. Mol Endocrinol 26:87–94CrossRefGoogle Scholar
  28. Kokabu S, Katagiri T, Yoda T, Rosen V (2012b) Role of Smad phosphatases in BMP-Smad signaling axis-induced osteoblast differentiation. J Oral Biosci 54:73–78CrossRefGoogle Scholar
  29. Kokabu S, Nojima J, Kanomata K, Ohte S, Yoda T, Fukuda T, Katagiri T (2010) Protein phosphatase magnesium-dependent 1A-mediated inhibition of BMP signaling is independent of Smad dephosphorylation. J Bone Miner Res 25:653–660CrossRefGoogle Scholar
  30. Kokabu S et al (2011) Suppression of BMP-Smad signaling axis-induced osteoblastic differentiation by small C-terminal domain phosphatase 1, a Smad phosphatase. Mol Endocrinol 25:474–481CrossRefGoogle Scholar
  31. Korupolu RV, Muenster U, Read JD, Vale W, Fischer WH (2008) Activin A/bone morphogenetic protein (BMP) chimeras exhibit BMP-like activity and antagonize activin and myostatin. J Biol Chem 283:3782–3790CrossRefGoogle Scholar
  32. Kretzschmar M, Doody J, Massague J (1997) Opposing BMP and EGF signalling pathways converge on the TGF-beta family mediator Smad1. Nature 389:618–622CrossRefGoogle Scholar
  33. Laine CM et al (2013) WNT1 mutations in early-onset osteoporosis and osteogenesis imperfecta. N Engl J Med 368:1809–1816CrossRefGoogle Scholar
  34. Li X, Zhang Y, Kang H, Liu W, Liu P, Zhang J, Harris SE, Wu D (2005) Sclerostin binds to LRP5/6 and antagonizes canonical Wnt signaling. J Biol Chem 280:19883–19887CrossRefGoogle Scholar
  35. Little RD et al (2002) A mutation in the LDL receptor-related protein 5 gene results in the autosomal dominant high-bone-mass trait. Am J Hum Genet 70:11–19CrossRefGoogle Scholar
  36. Mackay AM, Beck SC, Murphy JM, Barry FP, Chichester CO, Pittenger MF (1998) Chondrogenic differentiation of cultured human mesenchymal stem cells from marrow. Tissue Eng 4:415–428CrossRefGoogle Scholar
  37. Maeda K, Takahashi N, Kobayashi Y (2013) Roles of Wnt signals in bone resorption during physiological and pathological states. J Mol Med (Berl) 91:15–23CrossRefGoogle Scholar
  38. Massague J, Seoane J, Wotton D (2005) Smad transcription factors. Genes Dev 19:2783–2810CrossRefGoogle Scholar
  39. Moustakas A, Heldin CH (2005) Non-Smad TGF-beta signals. J Cell Sci 118:3573–3584CrossRefGoogle Scholar
  40. Mu Y, Gudey SK, Landstrom M (2012) Non-Smad signaling pathways. Cell Tissue Res 347:11–20CrossRefGoogle Scholar
  41. Mueller TD, Nickel J (2012) Promiscuity and specificity in BMP receptor activation. FEBS Lett 586:1846–1859CrossRefGoogle Scholar
  42. Nakao A et al (1997) Identification of Smad7, a TGFbeta-inducible antagonist of TGF-beta signalling. Nature 389:631–635CrossRefGoogle Scholar
  43. Nakashima K, Zhou X, Kunkel G, Zhang Z, Deng JM, Behringer RR, de Crombrugghe B (2002) The novel Zinc finger-containing transcription factor osterix is required for osteoblast differentiation and bone formation. Cell 108:17–29CrossRefGoogle Scholar
  44. Nakayama N, Duryea D, Manoukian R, Chow G, Han CY (2003) Macroscopic cartilage formation with embryonic stem-cell-derived mesodermal progenitor cells. J Cell Sci 116:2015–2028CrossRefGoogle Scholar
  45. Nishanian TG, Waldman T (2004) Interaction of the BMPR-IA tumor suppressor with a developmentally relevant splicing factor. Biochem Biophys Res Commun 323:91–97CrossRefGoogle Scholar
  46. Nojima J et al (2010) Dual roles of SMAD proteins in the conversion from myoblasts to osteoblastic cells by bone morphogenetic proteins. J Biol Chem 285:15577–15586CrossRefGoogle Scholar
  47. Ozkaynak E, Rueger DC, Drier EA, Corbett C, Ridge RJ, Sampath TK, Oppermann H (1990) OP-1 cDNA encodes an osteogenic protein in the TGF-beta family. EMBO J 9:2085–2093Google Scholar
  48. Papapoulos SE (2011) Targeting sclerostin as potential treatment of osteoporosis. Ann Rheum Dis 70(Suppl 1):i119–i122CrossRefGoogle Scholar
  49. Riley EH, Lane JM, Urist MR, Lyons KM, Lieberman JR (1996) Bone morphogenetic protein-2: biology and applications. Clin Orthop Relat Res 324:39–46Google Scholar
  50. Ruppert R, Hoffmann E, Sebald W (1996) Human bone morphogenetic protein 2 contains a heparin-binding site which modifies its biological activity. Eur J Biochem 237:295–302CrossRefGoogle Scholar
  51. Sampath TK et al (1990) Bovine osteogenic protein is composed of dimers of OP-1 and BMP-2A, two members of the transforming growth factor-beta superfamily. J Biol Chem 265:13198–13205Google Scholar
  52. Sampath TK, Reddi AH (1981) Dissociative extraction and reconstitution of extracellular matrix components involved in local bone differentiation. Proc Natl Acad Sci USA 78:7599–7603CrossRefGoogle Scholar
  53. Sanvitale CE et al (2013) A new class of small molecule inhibitor of BMP signaling. PLoS ONE 8:e62721CrossRefGoogle Scholar
  54. Seemann P et al (2009) Mutations in GDF5 reveal a key residue mediating BMP inhibition by NOGGIN. PLoS Genet 5:e1000747CrossRefGoogle Scholar
  55. Seemann P et al (2005) Activating and deactivating mutations in the receptor interaction site of GDF5 cause symphalangism or brachydactyly type A2. J Clin Invest 115:2373–2381CrossRefGoogle Scholar
  56. Shapiro F (2008) Bone development and its relation to fracture repair. The role of mesenchymal osteoblasts and surface osteoblasts. Eur Cell Mater 15:53–76Google Scholar
  57. Shukunami C, Ishizeki K, Atsumi T, Ohta Y, Suzuki F, Hiraki Y (1997) Cellular hypertrophy and calcification of embryonal carcinoma-derived chondrogenic cell line ATDC5 in vitro. J Bone Miner Res 12:1174–1188CrossRefGoogle Scholar
  58. Takada T, Katagiri T, Ifuku M, Morimura N, Kobayashi M, Hasegawa K, Ogamo A, Kamijo R (2003) Sulfated polysaccharides enhance the biological activities of bone morphogenetic proteins. J Biol Chem 278:43229–43235CrossRefGoogle Scholar
  59. Takase M, Imamura T, Sampath TK, Takeda K, Ichijo H, Miyazono K, Kawabata M (1998) Induction of Smad6 mRNA by bone morphogenetic proteins. Biochem Biophys Res Commun 244:26–29CrossRefGoogle Scholar
  60. Urist MR (1965) Bone: formation by autoinduction. Science 150:893–899CrossRefGoogle Scholar
  61. Urist MR, Strates BS (1971) Bone morphogenetic protein. J Dent Res 50:1392–1406CrossRefGoogle Scholar
  62. Valera E, Isaacs MJ, Kawakami Y, Izpisua Belmonte JC, Choe S (2010) BMP-2/6 heterodimer is more effective than BMP-2 or BMP-6 homodimers as inductor of differentiation of human embryonic stem cells. PLoS ONE 5:e11167CrossRefGoogle Scholar
  63. Walsh DW, Godson C, Brazil DP, Martin F (2010) Extracellular BMP-antagonist regulation in development and disease: tied up in knots. Trends Cell Biol 20:244–256CrossRefGoogle Scholar
  64. Wang EA, Rosen V, Cordes P, Hewick RM, Kriz MJ, Luxenberg DP, Sibley BS, Wozney JM (1988) Purification and characterization of other distinct bone-inducing factors. Proc Natl Acad Sci USA 85:9484–9488CrossRefGoogle Scholar
  65. Wosczyna MN, Biswas AA, Cogswell CA, Goldhamer DJ (2012) Multipotent progenitors resident in the skeletal muscle interstitium exhibit robust BMP-dependent osteogenic activity and mediate heterotopic ossification. J Bone Miner Res 27:1004–1017CrossRefGoogle Scholar
  66. Wozney JM, Rosen V, Celeste AJ, Mitsock LM, Whitters MJ, Kriz RW, Hewick RM, Wang EA (1988) Novel regulators of bone formation: molecular clones and activities. Science 242:1528–1534CrossRefGoogle Scholar
  67. Wu Q, Sun CC, Lin HY, Babitt JL (2012) Repulsive guidance molecule (RGM) family proteins exhibit differential binding kinetics for bone morphogenetic proteins (BMPs). PLoS ONE 7:e46307CrossRefGoogle Scholar
  68. Yamazaki M, Fukushima H, Shin M, Katagiri T, Doi T, Takahashi T, Jimi E (2009) Tumor necrosis factor alpha represses bone morphogenetic protein (BMP) signaling by interfering with the DNA binding of Smads through the activation of NF-kappaB. J Biol Chem 284:35987–35995CrossRefGoogle Scholar
  69. Yano K, Hoshino M, Ohta Y, Manaka T, Naka Y, Imai Y, Sebald W, Takaoka K (2009) Osteoinductive capacity and heat stability of recombinant human bone morphogenetic protein-2 produced by Escherichia coli and dimerized by biochemical processing. J Bone Miner Metab 27:355–363CrossRefGoogle Scholar
  70. Yang Y (2009) Skeletal morphogenesis during embryonic development. Crit Rev Eukaryot Gene Expr 19:197–218Google Scholar
  71. Zhang YE (2009) Non-Smad pathways in TGF-beta signaling. Cell Res 19:128–139CrossRefGoogle Scholar
  72. Zhao B et al (2006) Heparin potentiates the in vivo ectopic bone formation induced by bone morphogenetic protein-2. J Biol Chem 281:23246–23253CrossRefGoogle Scholar
  73. Zi Z, Chapnick DA, Liu X (2012) Dynamics of TGF-beta/Smad signaling. FEBS Lett 586:1921–1928CrossRefGoogle Scholar
  74. Zimmer J, Doelken SC, Horn D, Groppe JC, Shore EM, Kaplan FS, Seemann P (2012) Functional analysis of alleged NOGGIN mutation G92E disproves its pathogenic relevance. PLoS ONE 7:e35062CrossRefGoogle Scholar

Copyright information

© Springer International Publishing Switzerland 2015

Authors and Affiliations

  • Takenobu Katagiri
    • 1
    Email author
  • Sho Tsukamoto
    • 1
  • Kenji Osawa
    • 1
  • Shoichiro Kokabu
    • 2
  1. 1.Division of Pathophysiology, Research Center for Genomic MedicineSaitama Medical UniversityHidaka-shiJapan
  2. 2.Department of Oral and Maxillofacial Surgery, Faculty of MedicineSaitama Medical UniversityIruma-gunJapan

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