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Bone regeneration in critical size defects by cell-mediated BMP-2 gene transfer: a comparison of adenoviral vectors and liposomes

Gene Therapy volume 10, pages 1089–1098 (2003)Cite this article

Abstract

Large bone defects resulting from nonunion fractures or tumour resections are common clinical problems. Recent studies have shown bone morphogenetic protein-2 (BMP-2) gene transfer using adenoviral vectors to be a promising new therapeutic approach. However, comparative studies of different vectors are required to identify the optimal system for possible clinical trials. This study compares the use of liposome-mediated and adenoviral gene transfer for the generation of autologous BMP-2-producing bone marrow stromal cells (BMSC). Primary BMSC isolated from the rat femur were treated ex vivo with either an adenovirus or a liposome carrying human BMP-2 cDNA. The genetically modified cells were evaluated in vitro and transplanted into critical size defects in the rat mandible in vivo. BMSC treated with a reporter gene vector or untreated BMSC served as controls. The newly formed tissue was analysed by in situ hybridization, radiography and immunohistochemistry. Both groups of genetically modified cells produced BMP-2 for at least 2 weeks, and markers of new bone matrix such as osteopontin and osteocalcin were observed within 2 weeks following gene transfer. In the liposome group, the critical size defects were found completely healed at 6 weeks after the gene transfer, whereas the more efficient adenoviral gene transfer allowed for complete bone healing within 4 weeks. None of the three control groups showed bone healing, not even after 8 weeks. Thus, both liposome-mediated and adenoviral BMP-2 gene transfer to primary BMSC are suitable methods to achieve the healing of critical size bone defects in rats. As liposomes have proven sufficient for this purpose and offer several advantages over any other vector, such as ease of preparation, theoretically no limitation of the size of the DNA, and less immunological and safety problems, they may represent the best vector system for future clinical trials of bone regeneration by BMP-2 gene therapy.

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References

  1. Finkemeier CG . Bone-grafting and bone-graft substitutes. J Bone Joint Surg Am 2002; 84: 454–464.

    Article  Google Scholar 

  2. Wiltfang J et al. Degradation characteristics of alpha and beta tri-calcium-phosphate (TCP) in minipigs. J Biomed Mater Res 2002; 63: 115–121.

    Article  CAS  Google Scholar 

  3. Murphy WL, Mooney DJ . Controlled delivery of inductive proteins, plasmid DNA and cells from tissue engineering matrices. J Periodontal Res 1999; 34: 413–419.

    Article  CAS  Google Scholar 

  4. Perka C et al. Segmental bone repair by tissue-engineered periosteal cell transplants with bioresorbable fleece and fibrin scaffolds in rabbits. Biomaterials 2000; 21: 1145–1153.

    Article  CAS  Google Scholar 

  5. Wozney JM et al. Novel regulators of bone formation: molecular clones and activities. Science 1988; 242: 1528–1534.

    Article  CAS  Google Scholar 

  6. Wang EA et al. Recombinant human bone morphogenetic protein induces bone formation. Proc Natl Acad Sci 1990; 87: 2220–2224.

    Article  CAS  Google Scholar 

  7. Cook SD, Wolfe MW, Salkeld SL, Rueger DC . Effect of recombinant human osteogenic protein-1 on healing of segmental defects in non-human primates. J Bone Joint Surg Am 1995; 77: 734–750.

    Article  CAS  Google Scholar 

  8. Riley EH et al. Bone morphogenetic protein-2: biology and applications [Review]. Clin Orthop 1996; 324: 39–46.

    Article  Google Scholar 

  9. Cheng SL et al. In vitro and in vivo induction of bone formation using a recombinant adenoviral vector carrying the human BMP-2 gene. Calcif Tissue Int 2001; 68: 87–94.

    Article  CAS  Google Scholar 

  10. Mehrara B J et al. Adenovirus-mediated gene therapy of osteoblasts in vitro and in vivo. J Bone Miner Res 1999; 14: 1290–1301.

    Article  Google Scholar 

  11. Olmsted EA et al. Adenovirus-mediated BMP-2 expression in human bone marrow stromal cells. J Cell Biochem 2001; 82: 11–21.

    Article  CAS  Google Scholar 

  12. Riew KD et al. Induction of bone formation using a recombinant adenoviral vector carrying the human BMP-2 gene in a rabbit spinal fusion model. Calcif Tissue Int 1998; 63: 357–360.

    Article  CAS  Google Scholar 

  13. Lieberman JR et al. Regional gene therapy with a BMP-2-producing murine stromal cell line induces heterotopic and orthotopic bone formation in rodents. J Orthop Res 1998; 16: 330–339.

    Article  CAS  Google Scholar 

  14. Lieberman JR et al. The effect of regional gene therapy with bone morphogenetic protein-2 producing bone-marrow cells of repair of segmental femoral defects in rats. J Bone Jt Surg 1999; 81: 905–917.

    Article  CAS  Google Scholar 

  15. Musgrave DS et al. Adenovirus-mediated direct gene therapy with bone morphogenetic protein-2 produces bone. Bone 1999; 24: 541–547.

    Article  CAS  Google Scholar 

  16. Oakes DA, Lieberman JR . Osteoinductive applications of regional gene therapy: ex vivo gene transfer. Clin Orthop 2000; 379S:101–112.

    Article  Google Scholar 

  17. Baltzer AW et al. Genetic enhancement of fracture repair: healing of an experimental segmental defect by adenoviral transfer of the BMP-2 gene. Gene Ther 2000; 7: 734–739.

    Article  CAS  Google Scholar 

  18. Breitbart AS et al. Gene-enhanced tissue engineering: applications for bone healing using cultured periosteal cells transduced retrovirally with the BMP-7 gene. Ann Plast Surg 1999; 42: 488–495.

    Article  CAS  Google Scholar 

  19. Engstrand T et al. Transient production of bone morphogenetic protein 2 by allogeneic transplanted transduced cells induces bone formation. Hum Gene Ther 2000; 11: 205–211.

    Article  CAS  Google Scholar 

  20. Verma IM, Somia N . Gene therapy – promises, problems and prospects. Nature 1997; 389: 239–242.

    Article  CAS  Google Scholar 

  21. Wang EA, Israel DI, Kelly S, Luxenberg DP . Bone morphogenetic protein-2 causes commitment and differentiation in C3H10T1/2 and 3T3 cells. Growth Factors 1993; 9: 57–71.

    Article  CAS  Google Scholar 

  22. Gazit D et al. Engineered pluripotent mesenchymal cells integrate and differentiate in regenerating bone: a novel cell-mediated gene therapy. J Gene Med 1999; 1: 121–133.

    Article  CAS  Google Scholar 

  23. Gelse K et al. Fibroblast-mediated delivery of growth factor complementary DNA into mouse joints induces chondrogenesis but avoids the disadvantages of direct viral gene transfer. Arthritis Rheum 2001; 43: 1156–1164.

    Google Scholar 

  24. Gelse K et al. Articular cartilage repair in a rat cartilage defect model by cell-mediated gene therapy using growth factor-producing mesenchymal cells. Arthritis Rheum 2003; 48: 430–441.

    Article  CAS  Google Scholar 

  25. Kaban LB, Glowacki J . Induced osteogenesis in the repair of experimental bony defects in rats. J Dent Res 1981; 60: 1356.

    Article  CAS  Google Scholar 

  26. Kaban LB, Glowacki J, Murray JE . Repair of experimental bony defects in rats. Surg Forum 1979; 30: 519.

    CAS  PubMed  Google Scholar 

  27. Schmitz JP, Hollinger JO . The critical size defect as an experimental model for craniomandibulofacial nonunions. Clin Orthop 1986; 205: 299–308.

    Google Scholar 

  28. Lorente CA, Song BZ, Donoff RB . Healing of bony defects in the irradiated and unirradiated rat mandible. J Oral Maxillofac Surg 1992; 50: 1305–1309.

    Article  CAS  Google Scholar 

  29. Higuchi T et al. Bone regeneration by recombinant human bone morphogenetic protein-2 in rat mandibular defects. An experimental model of defect filling. J Periodontol 1999; 70: 1026–1031.

    Article  CAS  Google Scholar 

  30. Alden TD et al. The use of bone morphogenetic protein gene therapy in craniofacial bone repair. J Craniofac Surg 2000; 11: 24–30.

    Article  CAS  Google Scholar 

  31. Moutsatsos IK et al. Exogenously regulated stem cell-mediated gene therapy for bone regeneration. Mol Ther 2001; 3: 449–461.

    Article  CAS  Google Scholar 

  32. Lane JM et al. Bone marrow and recombinant human bone morphogenetic protein-2 in osseous repair. Clin Orthop 1999; 361: 216.

    Article  Google Scholar 

  33. Sellers RS et al. Repair of articular cartilage defects one year after treatment with recombinant human bone morphogenetic protein-2 (rhBMP-2). J Bone Jt Surg 2000; 82A: 151.

    Article  Google Scholar 

  34. Takushima A, Kitano Y, Harii K . Osteogenic potential of cultured periosteal cells in a distracted bone gap in rabbits. J Surg Res 1998; 78: 68–77.

    Article  CAS  Google Scholar 

  35. Perka C et al. Segmental bone repair by tissue-engineered periosteal cell transplants with bioresorbable fleece and fibrin scaffolds in rabbits. Biomaterials 2000; 21: 1145–1153.

    Article  CAS  Google Scholar 

  36. Hughes SS et al. Shared phenotypic expression of osteoblasts and chondrocytes in fracture callus. J Bone Miner Res 1995; 10: 533–544.

    Article  CAS  Google Scholar 

  37. Nishimura T, Simmons DJ, Mainous EG . The origin of bone formed by heterotopic periosteal autografts. J Oral Maxillofac Surg 1997; 55: 1265–1268.

    Article  CAS  Google Scholar 

  38. Harada K, Oida S, Sasaki S . BMP-2 can stimulate the bone marrow stromal cell to differentiate as osteoblast or chondroblast. Bone 1988; 9: 177–183.

    Article  CAS  Google Scholar 

  39. Harada K et al. BMP-2 induction and TGF-beta 1 modulation of rat periosteal cell chondrogenesis. J Cell Biochem 2001; 81: 284–294.

    Article  Google Scholar 

  40. Rowe NM et al. Rat mandibular distraction osteogenesis: Part I. Histologic and radiographic analysis. Plast Reconstr Surg 1998; 102: 2022–2032.

    Article  CAS  Google Scholar 

  41. Ikegame M et al. Tensile stress induces bone morphogenetic protein 4 in preosteoblastic and fibroblastic cells, which later differentiate into osteoblasts leading to osteogenesis in the mouse calvariae in organ culture. J Bone Mineral Res 2001; 16: 24–32.

    Article  CAS  Google Scholar 

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Acknowledgements

We thank Ms Heidemarie Heider for her expert technical assistance and Mrs Elisabeth Koppmann and Mrs Elisabeth Zimmermann for animal care. This work was supported by the Interdisciplinary Center for Clinical Research Grant D3 (to HS and KvdM) and ELAN Grant 01.04.12.1 from the University of Erlangen-Nuernberg.

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Authors and Affiliations

  1. Department of Oro-Maxillo-Facial Surgery, University of Erlangen-Nuremberg, Erlangen, Germany

    J Park, J Ries, F Kloss, J Wiltfang & F W Neukam

  2. Department of Experimental Medicine I, Nikolaus Fiebiger Center of Molecular Medicine, University of Erlangen-Nuremberg, Erlangen, Germany

    J Park, K Gelse, K von der Mark & H Schneider

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  1. J Park
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Park, J., Ries, J., Gelse, K. et al. Bone regeneration in critical size defects by cell-mediated BMP-2 gene transfer: a comparison of adenoviral vectors and liposomes. Gene Ther 10, 1089–1098 (2003). https://doi.org/10.1038/sj.gt.3301960

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