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. 2007 May 1;9(4):911–928. doi: 10.1111/j.1582-4934.2005.tb00388.x

Bone induction by recombinant human osteogenic protein‐1 (hOP‐1, BMP‐7) in the primate Papio ursinus with expression of mRNA of gene products of the TGF‐β superfamily

U Ripamonti 1,✉
PMCID: PMC6740134  PMID: 16364199

Abstract

Predictable bone induction in clinical contexts requires information on the expression and cross regulation of gene products of the transforming growth factor‐β (TGF‐β) superfamily elicited by single applications of each recombinant human bone morphogenetic/ostegenic proteins (BMPs/OPs). Using the calvarium and the rectus abdominis muscle of adult baboons Papio ursinus as a model for tissue induction and morphogenesis, this study investigated the induction of bone morphogenesis by γ‐irradiated hOP‐1 delivered by γ‐irradiated bovine insoluble collagenous bone matrix, the hOP‐1 osteogenic device, for bone induction in heterotopic and orthotopic sites of the primate Papio ursinus and the expression patterns of OP‐1, collagen type IV, BMP‐3 and TGFβ1mRNAs elicited by increasing single applications of doses of the hOP‐1 osteogenic devices (0.1, 0.5 and 2.5 mg hOP‐1/g of matrix) applied heterotopically in the rectus abdominis muscle and orthotopically in 48 calvarial defects of 12 adult baboons. Histology and histomorphometry on serial undecalcified sections prepared from the specimens harvested on day 15, 30 and 90 showed that all the doses of the hOP‐1 osteogenic device induced bone formation culminating in complete calvarial regeneration by day 90. Type IV collagen mRNA expression, a marker of angiogenesis, was strongly expressed in both heterotopic and orthotopic tissues. High levels of expression of OP‐1 mRNA demonstrated autoinduction of OP‐1 mRNAs. Expression levels of BMP‐3 mRNA varied from tissues induced in heterotopic vs. orthotopic sites with high expression in rapidly forming heterotopic ossicles together with high expression of type IV collagen mRNA. The temporal and spatial expressions of TGF‐β1 mRNA indicate a specific temporal transcriptional window during which expression of TGF‐β1 is mandatory for successful and optimal osteogenesis. The induction of bone by hOP‐1 in Papio ursinus develops as a mosaic structure with distinct spatial and temporal patterns of gene expression of members of the TGF‐β superfamily that singly, synergistically and synchronously initiate and maintain tissue induction and morphogenesis.

Keywords: bone morphogenetic proteins, osteogenic protein‐1, mRNA expression, TGF‐βsuperfamily members, redundancy, primates

References

  • 1. Ripamonti, U . Osteogenic proteins of the TGF‐β superfamily In: Henry HL, Norman AW, editors. Encyclopedia of Hormones. Austin Academic Press; 2004. p. 80–6. [Google Scholar]
  • 2. Ripamonti, U . Soluble, insoluble and geometric signals sculpt the architecture of mineralized bone. J Cell Mol Med. 2004; 8: 169–80. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 3. Ripamonti, U . Molecular signals in geometrical cues sculpt bone morphology. S Afr J Sci 2004; 100: 355–67. [Google Scholar]
  • 4. Wozney, JM , Rosen, V , Celeste, AJ , Mitsock, LM , Whitters, MJ , Kriz, RW , Hewick, RM , Wang, EA . Novel regulators of bone formation: Molecular clones and activities. Science 1988; 242: 1528–34. [DOI] [PubMed] [Google Scholar]
  • 5. Reddi, AH . Bone and cartilage differentiation. Curr Opin Genet Dev. 1994; 4: 737–44. [DOI] [PubMed] [Google Scholar]
  • 6. Reddi, AH . Role of morphogenetic proteins in skeletal tissue engineering and regeneration. Nature Biotechnol. 1998; 16: 247–52. [DOI] [PubMed] [Google Scholar]
  • 7. Reddi, AH . Morphogenesis and tissue engineering of bone and cartilage: Inductive signals, stem cells, and biomimetic biomaterials. Tissue Eng. 2001; 6: 351–59. [DOI] [PubMed] [Google Scholar]
  • 8. Ripamonti, U , Ramoshebi, LN , Matsaba, T , Tasker, J , Crooks, J , Teare, J . Bone induction by BMPs/OPs and related family members in primates. The critical role of delivery systems. J Bone Joint Surg. 2001; 83‐A: S1: 116–27. [PubMed] [Google Scholar]
  • 9. Ripamonti, U , Ramoshebi, LN , Patton, J , Matsaba, T , Teare, J , Renton, L . Soluble signals and insoluble substrata: Novel molecular cues instructing the induction of bone In Massaro EJ. and Rogers JM. (Eds.) The Skeleton. Humana Press, Chapter 15, 2004. p. 217–27. [Google Scholar]
  • 10. Reddi, AH . Bone morphogenetic proteins: An unconventional approach to isolation of first mammalian morphogens. Cytokine Growth Factor Rev 1997; 8: 11–20. [DOI] [PubMed] [Google Scholar]
  • 11. Sampath, TK , Rashka, KE , Doctor, JS , Tucker, RF , Hoffmann, FM . Drosophila TGF‐β superfamily proteins induce endochondral bone formation in mammals. Proc Natl Acad Sci USA 1993; 90: 6004–8. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 12. Sampath, TK , Maliakal, JC , Hauschka, PV , Jones, WK , Sasak, H , Tucker, RF , White, KH , Coughlin, JE , Tucker, MM , Pang, RH , et al. Recombinant human osteogenic protein‐1 (hOP‐1) induces new bone formation in vivo with a specific activity comparable with natural bovine osteogenic protein and stimulates osteoblast proliferation and differentiation in vitro . J Biol Chem 1992; 267: 20352–62. [PubMed] [Google Scholar]
  • 13. Ripamonti, U , van den Heever, B , Crooks, J , Rueger, DC , Reddi, AH . Long‐term evaluation of bone formation by osteogenic protein 1 in the baboon and relative efficacy of bone‐derived bone morphogenetic proteins delivered by irradiated xenogeneic collagenous matrices. J Bone Miner Res 2000; 15: 1798–809. [DOI] [PubMed] [Google Scholar]
  • 14. Sampath, TK , Reddi, AH . Dissociative extraction and reconstitution of extracellular matrix components involved in local bone differentiation. Proc Natl Acad Sci USA 1981; 78: 7599–603. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 15. Wang, H , Springer, IN , Schildberg, H , Ludwig, K , Rueger, DC , Terheyden, H . Carboxymethylcellulose‐stabilized collagenous rhOP‐1 device ‐ a novel carrier biomaterial for the repair of mandibular continuity defects. J Biomed Mater Res 2004; 68: 219–26. [DOI] [PubMed] [Google Scholar]
  • 16. Duneas, N , Crooks, J , Ripamonti, U . Transforming growth factor‐β1: Induction of bone morphogenetic protein genes expression during endochondral bone formation in the baboon, and synergistic interaction with osteogenic protein‐1 (BMP‐7). Growth Factors 1998; 15: 259–77. [DOI] [PubMed] [Google Scholar]
  • 17. Ripamonti, U , Crooks, J , Matsaba, T , Tasker, J . Induction of endochondral bone formation by recombinant human transforming growth factor‐β2 in the baboon (Papio ursinus). Growth Factors 2000; 17: 269–85. [DOI] [PubMed] [Google Scholar]
  • 18. de Rynk, R , Jarret, JA , Chen, EY , Eaton, EH , Bell, JR , Assoian, RK , Roberts, AB , Sporn, MB , Goeddel, DV . Human transforming growth factor‐β complementary DNA sequence and expression in normal and transformed cells. Nature 1985; 316: 701–5. [DOI] [PubMed] [Google Scholar]
  • 19. Hostikka, SL , Tryggvason, K . The complete primary structure of the α2 chain of the human type IV collagen and comparison with the α (IV) chain. J Biol Chem 1988; 263: 19488–93. [PubMed] [Google Scholar]
  • 20. Özkaynak, E , Rueger, DC , Drier, EA , Corbett, C , Ridge, RJ , Samapth, T , Oppermann, H . OP‐1 cDNA encodes an osteogenic protein in the TGF‐β family. Embo J Org 1990; 9: 2085–93. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 21. Helder, MN , Özkaynak, E , Sampath, KT , Luyten, FP , Latin, V , Oppermann, H , Vukicevic, S . Expression of osteogenic protein‐1 (bone morhogenetic protein‐7) in human mouse development. J Histochem Cytochem 1995; 43: 1035–44. [DOI] [PubMed] [Google Scholar]
  • 22. Ripamonti, U . Bone induction in nonhuman primates. An experimental study on the baboon (Papio ursinus). Clin Orthop Rel Res 1991; 269: 284–94. [PubMed] [Google Scholar]
  • 23. Public Service Department . National Code for Animal Use in Research Education, Diagnosis and Testing of Drugs and Related Substances in South Africa. Public Service Department, Pretoria , South Africa 1990. [Google Scholar]
  • 24. Ripamonti, U , Ma, S , Cunningham, N , Yates, L , Reddi, AH . Initiation of bone regeneration in adult baboons by osteogenin, a bonemorphogenetic protein. Matrix 1992; 12: 369–80. [DOI] [PubMed] [Google Scholar]
  • 25. Ripamonti, U , Ma, S , Cunningham, N , Yeates, L , Reddi, AH . Reconstruction of bone‐bone marrow organ by osteogenin, a bone morphogenetic protein, and demineralized bone matrix in calvarial defects of adult primates. Plast Reconstr Surg 1993; 91: 263–67. [DOI] [PubMed] [Google Scholar]
  • 26. Ripamonti, U , van den Heever, B , Sampath, TK , Tucker, MM , Rueger, DC , Reddi, AH . Complete regeneration of bone in the baboon by recombinant human osteogenic protein‐1 (hOP‐1, bone morphogenetic protein‐7). Growth Factors 1996; 13: 273–89. [DOI] [PubMed] [Google Scholar]
  • 27. Parfitt, AM . Stereologic basis of bone histomorphometry; theory of quantitative microscopy and reconstruction of the third dimension In: Recker RR. (ed). Bone Histomorphometry: Techniques and Interpretation. CRC Press, Boca Raton , FL USA p 53–87. [Google Scholar]
  • 28. Parfitt, AM , Drezner, MK , Gliorieux, FH , Kanis, JA , Malluche, H , Meunier, PJ , Ott, SM , Recker, RR . Bone histomorphometry: Standardization of nomenclature, symbols, and units. J Bone Miner Res 1987; 2: 595–610. [DOI] [PubMed] [Google Scholar]
  • 29. Ripamonti, U , Duneas, N , van den Heever, B , Bosch, C , Crooks, J . Recombinant transforming growth factor‐β1 induces endochondral bone in the baboon and synergizes with recombinant osteogenic protein‐1 (bone morphogenetic protein‐7) to initiate rapid bone formation. J Bone Miner Res 1997; 2: 1584–95. [DOI] [PubMed] [Google Scholar]
  • 30. Statistical Analysis System SAS/STATS User's Guide, version 6, 4th ed. Vol 1 SAS Institute, Inc., Cary , NC , USA , 1989. p. 209–44. [Google Scholar]
  • 31. Roman‐Roman, S , Garcia, T , Jackson, A , Theilhabr, J , Rawadi, G , Connolly, T , Spinella‐Jaegle, S , Kawai, S , Courtois, B , Bushnell, S , Auberval, M , Call, K , Baron, R . Identification of genes regulated during osteoblastic differentiation by genome‐wide expression analysis of mouse calvaria primary osteoblasts in vitro Bone 2003; 32: 474–82. [DOI] [PubMed] [Google Scholar]
  • 32. Cho, T , Gerstenfeld, L , Einhorn, T . Differential temporal expression of multiple members of TGF β superfamily during murine bone healing. 47th Annual Meeting, Orthopaedic Research Society, February 25–28, San Francisco, California. 2001.
  • 33. Spector, J.A , Luchs, J.S. , Mehrara, B.J , Greenwald, JA , Smith, LP , Longaker, MT . Expression of bone morphogenetic proteins during membranous bone healing. Plast Reconstr Surg 2001; 107: 124–34. [DOI] [PubMed] [Google Scholar]
  • 34. Yeh, LC , Unda, R , Lee, JC . Osteogenic protein‐1 differentially regulates the mRNA expression of bone morphogenetic proteins and their receptors in primary cultures of osteoblasts. J Cell Physiol 2000; 185: 87–97. [DOI] [PubMed] [Google Scholar]
  • 35. Bahamonde, M. E , Lyons, K. M . BMP3: To be or not to be a BMP. J Bone Joint Surg 2001; 83‐A: S1: 56–62. [PubMed] [Google Scholar]
  • 36. Chen, D , Harris, MA , Rossini, G , Dunstan, CR , Dallas, SL , Feng, JQ , Mundy, GR , Harris, SE . 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. [DOI] [PubMed] [Google Scholar]
  • 37. Honda, Y , Knutsen, R , Strong, DD , Sampath, TK , Baykink, DJ , Mohan, S . Osteogenic protein‐l stimulates mRNA levels of BMP‐6 and decreases mRNA levels of BMP‐2 and −4 in human osteosarcoma cells. Calcif Tissue Int 1997; 60: 297–301. [DOI] [PubMed] [Google Scholar]
  • 38. Ripamonti, U , Herbst, N‐N , Ramoshebi, KN . Bone morphogenetic proteins in craniofacial and periodontal tissue engineering: Experimental studies in the non‐human primate Papio ursinus . Cytokine Growth Factors Rev 2005; 16: 357–68. [DOI] [PubMed] [Google Scholar]
  • 39. Ramoshebi, LN , Ripamonti, U . Osteogenic protein‐l, a bone morphognetic protein, induces angiogenesis in the chick chorioallantoic membrane and synergizes wiht basic fibroblast growth factor and transforming growth factor‐ β. Anat Record. 2000; 259: 97–107. [DOI] [PubMed] [Google Scholar]
  • 40. Folkman, J , Klagsbrun, M , Sasse, J , Wadzinski, M , Ingber, D , Wlodavsky, I . A heparin‐bind angiognic protein‐basic fibroblast growth factor‐is stored within basement membranes. Am J Pathol. 1988; 130: 393–400. [PMC free article] [PubMed] [Google Scholar]
  • 41. Wlodavsky, I , Folkman, J , Sullivan, R , Fridman, R , Ishai‐Michaeli, R , Klagsbrun, M . Endothelial cell‐dervied basic fibroblast growth factor. Synthesis and deposition into subendothelial extracellular matrix. proc Natl Acad Sci USA. 1987; 84: 2292–96. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 42. Paralkar, VM , Nandekar, AKN , Pointer, RH , Kleinman, HK , Reddi, AH . Interaction of osteogenin, a heparin binding bone morphogenetic protein, with type IV collagen. J Biol Chem 1990; 265: 1781–84. [PubMed] [Google Scholar]
  • 43. Paralkar, VM , Vukicevic, S , Reddi, AH . Transforming growth factor β typel binds to collagen type IV of basement membrane matrix: Implication for development. Dev Biol. 1991; 143: 303–8. [DOI] [PubMed] [Google Scholar]
  • 44. de Ckers, MML , Rutger, L , van Bezooijen, RL , van der Horst, G , Hoogendam, J , van der Bent, C , Papapoulus, Löwik CWGM . Bone morphogenetic proteins stimulate angiogenesis through osteoblast‐derived vasculalr endothelial growth factor A. Endocrinology 143: 1545–53. [DOI] [PubMed] [Google Scholar]
  • 45. Tabibzadeh, S , Kothapalli, R , Buyuksal, I . Distinct tumor specific expression of TGFβ4 (ebaf), an novel human geme of the TGF‐β superfamily. Front Biosci. 1997; 2: 18–25. [DOI] [PubMed] [Google Scholar]
  • 46. Kothapalli, R , Buyukasal, I , Wu, S‐Q , Chegini, N , Tabibzadeh, S . Detection of ebaf, a novel human gene of the transforming growth factor β superfamily. Association of gene expression with endometrial bleeding. J Clin Invest. 1997; 99: 2342–50. [DOI] [PMC free article] [PubMed] [Google Scholar]

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