Are Endogenous BMPs Necessary for Bone Healing during Distraction Osteogenesis? (original) (raw)

Abstract

Previous reports suggest the application of exogenous BMPs can accelerate bone formation during distraction osteogenesis (DO). However, there are drawbacks associated with the use of exogenous BMPs. A possible alternative to the use of exogenous BMPs is to upregulate the expression of endogenous BMPs. Since DO results in spontaneously generated de novo bone formation in a uniform radiographic, histological, and biomechanical temporal sequence, a genetically engineered model lacking endogenous BMP2 should have measurable deficits in bone formation at different time points. We performed DO on BMP2 fl/+ and BMP2 fl/+ cre mice using a miniature Ilizarov fixator. Distracted samples were collected at various time points and analyzed using Real Time-quantitative PCR, μCT, radiology, immunohistochemistry, histology, and biomechanical testing. Immunohistochemical studies of 34-day heterozygous samples showed reduced expression of BMP2, BMP7, BMPR1a, ACTR1, and ACTR2b. μCT analysis of 51-day heterozygous samples revealed a decrease in trabecular number and increase in trabecular separation. Biomechanical testing of 51-day heterozygous samples revealed decreased stiffness and increased ultimate displacement. Radiological analysis showed the heterozygotes contained a decreased bone fill score at 17, 34, and 51 days. These data suggest endogenous BMPs are important for bone healing and manipulating endogenous BMPs may help accelerate bone consolidation during DO.

Access this article

Log in via an institution

Subscribe and save

Buy Now

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

Instant access to the full article PDF.

Similar content being viewed by others

References

  1. Abbaspour A, Takata S, Sairyo K, Katoh S, Yukata K, Yasui N. Continuous local infusion of fibroblast growth factor-2 enhances consolidation of the bone segment lengthened by distraction osteogenesis in rabbit experiment. Bone. 2008;42:98–106.
    Article CAS PubMed Google Scholar
  2. Axelrad T, Steen B, Lowenberg D, Creevy W, Einhorn T. Heterotopic ossification after the use of commercially available recombinant human bone morphogenetic proteins in four patients. J Bone Joint Surg Br. 2008;90:1617–1622.
    Article CAS PubMed Google Scholar
  3. Birch J, Samchukov M. Use of the Ilizarov method to correct lower limb deformities in children and adolescents. J Am Acad Orthop Surg. 2004;12:144–154.
    PubMed Google Scholar
  4. Casap N, Venezia N, Wilensky A, Samuni Y. VEGF facilitates periosteal distraction-induced osteogenesis in rabbits: a micro-computerized tomography study. Tissue Eng Part A. 2008;14:247–253.
    Article CAS PubMed Google Scholar
  5. Colburn N, Zaal K, Wang F, Tuan R. A role for/T cells in a mouse model of fracture healing. Arthritis Rheum. 2009;60:1694–1703.
    Article CAS PubMed Google Scholar
  6. Fowlkes J, Thrailkill K, Liu L, Wahl E, Bunn R, Cockrell G, Perrien D, Aronson J, Lumpkin Jr C. Effects of systemic and local administration of recombinant human IGF-I (rhIGF-I) on de novo bone formation in an aged mouse model. J Bone Miner Res. 2006;21:1359–1366.
    Article CAS PubMed Google Scholar
  7. Friedlaender G, Perry C, Cole J, Cook S, Cierny G, Muschler G, Zych G, Calhoun J, LaForte A, Yin S. Osteogenic protein-1 (bone morphogenetic protein-7) in the treatment of tibial nonunions. J Bone Joint Surg Am. 2001;83:S151–S158.
    PubMed Google Scholar
  8. Giannoudis P, Kanakaris N, Einhorn T. Interaction of bone morphogenetic proteins with cells of the osteoclast lineage: review of the existing evidence. Osteoporos Int. 2007;18:1565–1581.
    Article CAS PubMed Google Scholar
  9. Govender S, Csimma C, Genant H, Valentin-Opran A, Amit Y, Arbel R, Aro H, Atar D, Bishay M, Börner M, Chiron P, Choong P, Cinats J, Courtenay B, Feibel R, Geulette B, Gravel C, Haas N, Raschke M, Hammacher E, van der Velde D, Hardy P, Holt M, Josten C, Ketterl RL, Lindeque B, Lob G, Mathevon H, McCoy G, Marsh D, Miller R, Munting E, Oevre S, Nordsletten L, Patel A, Pohl A, Rennie W, Reynders P, Rommens PM, Rondia J, Rossouw WC, Daneel PJ, Ruff S, Rüter A, Santavirta S, Schildhauer TA, Gekle C, Schnettler R, Segal D, Seiler H, Snowdowne RB, Stapert J, Taglang G, Verdonk R, Vogels L, Weckbach A, Wentzensen A, Wisniewski T. BMP-2 Evaluation in Surgery for Tibial Trauma (BESTT) Study Group. Recombinant human bone morphogenetic protein-2 for treatment of open tibial fractures: a prospective, controlled, randomized study of four hundred and fifty patients. J Bone Joint Surg Am. 2002;84:2123–2134.
    Google Scholar
  10. Haque T, Hamade F, Alam N, Kotsiopriftis M, Lauzier D, St-Arnaud R, Hamdy RC. Characterizing the BMP pathway in a wild type mouse model of distraction osteogenesis. Bone. 2008;42:1144–1153.
    Article CAS PubMed Google Scholar
  11. Haque T, Mandu-Hrit M, Rauch F, Lauzier D, Tabrizian M, Hamdy RC. Immunohistochemical localization of bone morphogenetic protein-signaling Smads during long-bone distraction osteogenesis. J Histochem Cytochem. 2006;54:407–415.
    Article CAS PubMed Google Scholar
  12. Hsu W, Wang J. The use of bone morphogenetic protein in spinal fusion. Spine J. 2008;8:419–425.
    Article PubMed Google Scholar
  13. Hu J, Li J, Wang D, Buckley M, Agarwal S. Differences in mandibular distraction osteogenesis after corticotomy and osteotomy. Int J Oral Maxillofac Surg. 2002;31:185–189.
    Article CAS PubMed Google Scholar
  14. Hu J, Qi M, Zou S, Li J, Luo E. Callus formation enhanced by BMP-7 ex vivo gene therapy during distraction osteogenesis in rats. J Orthop Res. 2007;25:241–251.
    Article CAS PubMed Google Scholar
  15. Ilizarov G. The tension-stress effect on the genesis and growth of tissues. I: The infleunces of stability of fixation and soft-tissue preservation. Clin Orthop Relat Res. 1989;238:249–281.
    PubMed Google Scholar
  16. Ilizarov G. The tension-stress effect on the genesis and growth of tissues: Part II. The influence of the rate and frequency of distraction. Clin Orthop Relat Res. 1989;239:263–285.
    PubMed Google Scholar
  17. Lammens J, Liu Z, Luyten F. Bone morphogenetic protein signaling in the murine distraction osteogenesis model. Acta Orthop Belg. 2009;75:94–102.
    PubMed Google Scholar
  18. Lammens J, Nijs J, Schepers E, Ectors N, Lismont D, Verduyckt B. The effect of bone morphogenetic protein-7 (OP-1) and demineralized bone matrix (DBM) in the rabbit tibial distraction model. Acta Orthop Belg. 2009;75:103–109.
    PubMed Google Scholar
  19. Lloyd S, Yuan Y, Kostenuik P, Ominsky M, Lau A, Morony S, Stolina M, Asuncion F, Bateman T. Soluble RANKL induces high bone turnover and decreases bone volume, density, and strength in mice. Calcif Tissue Int. 2008;82:361–372.
    Article CAS PubMed Google Scholar
  20. Mandu-Hrit M, Haque T, Lauzier D, Kotsiopriftis M, Rauch F, Tabrizian M, Henderson J, Hamdy RC. Early injection of OP-1 during distraction osteogenesis accelerates new bone formation in rabbits. Growth Factors. 2006;24:172–183.
    Article CAS PubMed Google Scholar
  21. Moore D, Ehrlich M, McAllister S, Machan J, Hart C, Voigt C, Lesieur-Brooks A, Webber E. Recombinant human platelet-derived growth factor-BB augmentation of new-bone formation in a rat model of distraction osteogenesis. J Bone Joint Surg Am. 2009;91:1973–1984.
    Article PubMed Google Scholar
  22. Namdari S, Wei L, Moore D, Chen Q. Reduced limb length and worsened osteoarthritis in adult mice after genetic inhibition of p38 MAP kinase activity in cartilage. Arthritis Rheum. 2008;58:3520–3529.
    Article CAS PubMed Google Scholar
  23. Okamoto M, Murai J, Yoshikawa H, Tsumaki N. Bone morphogenetic proteins in bone stimulate osteoclasts and osteoblasts during bone development. J Bone Miner Res. 2006;21:1022–1033.
    Article CAS PubMed Google Scholar
  24. Ozkan K, Eralp L, Kocaoglu M, Ahishali B, Bilgic B, Mutlu Z, Turker M, Ozkan F. The effect of transforming growth factor 1 (TGF-1) on the regenerate bone in distraction osteogenesis. Growth Factors. 2007;25:101–107.
    Article CAS PubMed Google Scholar
  25. Paley D. Problems, obstacles, and complications of limb lengthening by the Ilizarov technique. Clin Orthop Rel Res 1990;250:81–104.
    Google Scholar
  26. Rengachary S. Bone morphogenetic proteins: basic concepts. Neurosurg Focus. 2002;13:e2.
    Google Scholar
  27. Shimizu T, Jayawardana B, Nishimoto H, Kaneko E, Tetsuka M, Miyamoto A. Involvement of the bone morphogenetic protein/receptor system during follicle development in the bovine ovary: Hormonal regulation of the expression of bone morphogenetic protein 7 (BMP-7) and its receptors (ActRII and ALK-2). Mol Cell Endocrinol. 2006;249:78–83.
    CAS PubMed Google Scholar
  28. Tay B, Le A, Gould S, Helms J. Histochemical and molecular analyses of distraction osteogenesis in a mouse model. J Orthop Res. 1998;16:636–642.
    Article CAS PubMed Google Scholar
  29. Troulis M, Coppe C, O’Neil MJ, Kaban LB. Ultrasound: assessment of the distraction osteogenesis wound in patients undergoing mandibular lengthening. J Oral Maxillofac Surg. 2003;61:1144–1149.
    Article PubMed Google Scholar
  30. Tsuji K, Bandyopadhyay A, Harfe B, Cox K, Kakar S, Gerstenfeld L, Einhorn T, Tabin C, Rosen V. BMP2 activity, although dispensable for bone formation, is required for the initiation of fracture healing. Nat Genet. 2006;38:1424–1429.
    Article CAS PubMed Google Scholar
  31. Vaibhav B, Nilesh P, Vikram S, Anshul C. Bone morphogenic protein and its application in trauma cases: A current concept update. Injury. 2007;38:1227–1235.
    Article PubMed Google Scholar
  32. Wysocki R, Cohen M. Ectopic ossification of the triceps muscle after application of bone morphogenetic protein-7 to the distal humerus for recalcitrant nonunion: a case report. J Hand Surg Am. 2007;32:647–650.
    Article PubMed Google Scholar
  33. Zhang H. Mice deficient for BMP2 are nonviable and have defects in amnion/chorion and cardiac development. Development. 1996;122:2977–2986.
    CAS PubMed Google Scholar

Download references

Acknowledgements

We thank members of Dr. St-Arnaud’s lab, Fares Hamade, Tasima Haque, Maria Kotsioprifitis, and Noémi Dahan for assisting with the study, Dr. Vicki Rosen (Harvard School of Medicine, Boston, MA) for the conditional BMP2 knockout mice, Guylaine Bedard and Mark Lepik for help with figures, and the McGill Bone Centre for their assistance with the radiology/μCT and biomechanical testing analysis.

Author information

Authors and Affiliations

  1. Division of Orthopaedics, Shriners Hospital for Children, Montréal, QC, Canada
    Norine Alam MS
  2. Genetics Unit, Shriners Hospital for Children, McGill University, Montréal, QC, Canada
    René St-Arnaud PhD
  3. Division of Orthopaedics, Montréal Children’s Hospital, Montréal, QC, Canada
    Dominique Lauzier AS
  4. Department of Developmental Biology, Harvard School of Dental Medicine, Boston, MA, USA
    Vicki Rosen PhD
  5. Division of Orthopaedics, Shriners Hospital and Montreal Children Hospital, McGill University, 1529 Cedar Avenue, Montréal, QC, H3G 1A6, Canada
    Reggie C. Hamdy MD, FRCSC

Authors

  1. Norine Alam MS
    You can also search for this author inPubMed Google Scholar
  2. René St-Arnaud PhD
    You can also search for this author inPubMed Google Scholar
  3. Dominique Lauzier AS
    You can also search for this author inPubMed Google Scholar
  4. Vicki Rosen PhD
    You can also search for this author inPubMed Google Scholar
  5. Reggie C. Hamdy MD, FRCSC
    You can also search for this author inPubMed Google Scholar

Corresponding author

Correspondence toReggie C. Hamdy MD, FRCSC.

Additional information

Each author certifies that he or she has no commercial associations (eg, consultancies, stock ownerships, equity interest, patent/licensing arrangements, etc) that might pose a conflict of interest in connection with the submitted article. This work was supported by Shriners of North America operating grant no. 8700.

Each author certifies that his or her institution has approved the animal protocol for this investigation and that all investigations were conducted in conformity with ethical principles of research.

This work was performed at the Shriners Hospital for Children and the Montréal Children’s Hospital of McGill University, Montréal, QC, Canada.

About this article

Cite this article

Alam, N., St-Arnaud, R., Lauzier, D. et al. Are Endogenous BMPs Necessary for Bone Healing during Distraction Osteogenesis?.Clin Orthop Relat Res 467, 3190–3198 (2009). https://doi.org/10.1007/s11999-009-1065-6

Download citation

Keywords