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Drilled Hole Defects in Mouse Femur as Models of Intramembranous Cortical and Cancellous Bone Regeneration

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Abstract

In order to identify pertinent models of cortical and cancellous bone regeneration, we compared the kinetics and patterns of bone healing in mouse femur using two defect protocols. The first protocol consisted of a 0.9-mm-diameter through-and-through cortical hole drilled in the mid-diaphysis. The second protocol was a 0.9-mm-diameter, 1-mm-deep perforation in the distal epimetaphyseal region, which destroyed part of the growth plate and cancellous bone. Bone healing was analyzed by ex vivo micro-computerized X-ray tomography and histology. In the diaphysis, the cortical gap was bridged with woven bone within 2 weeks. This newly formed bone was rapidly remodeled into compact cortical bone, which showed characteristic parameters of intact cortex 4 weeks after surgery. In the epimetaphysis, bone formation was initiated at the deepest region of the defect and spread slowly toward the cortical gap. In this position, newly formed bone quickly adopted the characteristics of trabecular bone, whereas a thin compact wall was formed at its external border, which reached the density of intact cortical bone but failed to bridge the cortical gap even 13 weeks after surgery. This comparative study indicates that the diaphyseal defect is a model of cortical bone healing and that the epimetaphyseal defect is a model of cancellous bone repair. These models enable experimental genetics studies to investigate the cellular and molecular mechanisms of spontaneous cortical and cancellous bone repair and may be useful for pharmacological studies.

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References

  1. Shapiro F (2008) Bone development and its relation to fracture repair. The role of mesenchymal osteoblasts and surface osteoblasts. Eur Cell Mater 15:53–76

    CAS  PubMed  Google Scholar 

  2. Alford AI, Hankenson KD (2006) Matricellular proteins: extracellular modulators of bone development, remodeling, and regeneration. Bone 38:749–757

    Article  CAS  PubMed  Google Scholar 

  3. Schindeler A, McDonald MM, Bokko P, Little DG (2008) Bone remodeling during fracture repair: the cellular picture. Semin Cell Dev Biol 19:459–466

    Article  CAS  PubMed  Google Scholar 

  4. Choi P, Ogilvie C, Thompson Z, Miclau T, Helms JA (2004) Cellular and molecular characterization of a murine non-union model. J Orthop Res 22:1100–1107

    Article  CAS  PubMed  Google Scholar 

  5. Wahl EC, Liu L, Perrien DS, Aronson J, Hogue WR, Skinner RA, Hidestrand M, Ronis MJ, Badger TM, Lumpkin CK Jr (2006) A novel mouse model for the study of the inhibitory effects of chronic ethanol exposure on direct bone formation. Alcohol 39:159–167

    Article  CAS  PubMed  Google Scholar 

  6. Uusitalo H, Rantakokko J, Ahonen M, Jamsa T, Tuukkanen J, KaHari V, Vuorio E, Aro HT (2001) A metaphyseal defect model of the femur for studies of murine bone healing. Bone 28:423–429

    Article  CAS  PubMed  Google Scholar 

  7. Campbell TM, Wong WT, Mackie EJ (2003) Establishment of a model of cortical bone repair in mice. Calcif Tissue Int 73:49–55

    Article  CAS  PubMed  Google Scholar 

  8. Cui Q, Xiao Z, Li X, Saleh KJ, Balian G (2006) Use of genetically engineered bone-marrow stem cells to treat femoral defects: an experimental study. J Bone Joint Surg Am 88 Suppl 3:167–172

    Article  PubMed  Google Scholar 

  9. Nagashima M, Sakai A, Uchida S, Tanaka S, Tanaka M, Nakamura T (2005) Bisphosphonate (YM529) delays the repair of cortical bone defect after drill-hole injury by reducing terminal differentiation of osteoblasts in the mouse femur. Bone 36:502–511

    Article  CAS  PubMed  Google Scholar 

  10. Baron R, Vignery A, Neff L, Silvergate A, Santa Maria A (1983) Processing of undecalcified bone specimens for bone histomorphometry. In: RR B (ed) Bone histomorphometry: techniques and interpretations. CRC Press, Boca Raton, pp 13–35

  11. Schenk RK, Olah AJ, Herrmann W (1984) Preparation of calcified tissues for light microscopy. In: Dickson GR (ed) Methods of calcified tissue preparation. Elsevier, Amsterdam, pp 1–56

  12. Goldner J (1938) A modification of the Masson Trichrome technique for routine laboratory purpose. Am J Pathol 14:237–243

    Google Scholar 

  13. Makela EA, Vainionpaa S, Vihtonen K, Mero M, Rokkanen P (1988) The effect of trauma to the lower femoral epiphyseal plate. An experimental study in rabbits. J Bone Joint Surg Br 70:187–191

    CAS  PubMed  Google Scholar 

  14. Cottalorda J, Jouve JL, Bollini G, Panuel M, Guisiano B, Jimeno MT (1996) Epiphyseal distraction and centrally located bone bar: an experimental study in the rabbit. J Pediatr Orthop 16:664–668

    CAS  PubMed  Google Scholar 

  15. Lee MA, Nissen TP, Otsuka NY (2000) Utilization of a murine model to investigate the molecular process of transphyseal bone formation. J Pediatr Orthop 20:802–806

    CAS  PubMed  Google Scholar 

  16. Xian CJ, Zhou FH, McCarty RC, Foster BK (2004) Intramembranous ossification mechanism for bone bridge formation at the growth plate cartilage injury site. J Orthop Res 22:417–426

    Article  PubMed  Google Scholar 

  17. Zhou FH, Foster BK, Sander G, Xian CJ (2004) Expression of proinflammatory cytokines and growth factors at the injured growth plate cartilage in young rats. Bone 35:1307–1315

    Article  CAS  PubMed  Google Scholar 

  18. Colnot C (2009) Skeletal cell fate decisions within periosteum and bone marrow during bone regeneration. J Bone Miner Res 24:274–282

    Article  PubMed  Google Scholar 

  19. Bilic-Curcic I, Kalajzic Z, Wang L, Rowe DW (2005) Origins of endothelial and osteogenic cells in the subcutaneous collagen gel implant. Bone 37:678–687

    Article  CAS  PubMed  Google Scholar 

  20. Suva LJ, Seedor JG, Endo N, Quartuccio HA, Thompson DD, Bab I, Rodan GA (1993) Pattern of gene expression following rat tibial marrow ablation. J Bone Miner Res 8:379–388

    Article  CAS  PubMed  Google Scholar 

  21. Grellier M, Granja PL, Fricain JC, Bidarra SJ, Renard M, Bareille R, Bourget C, Amedee J, Barbosa MA (2009) The effect of the co-immobilization of human osteoprogenitors and endothelial cells within alginate microspheres on mineralization in a bone defect. Biomaterials 30:3271–3278

    Article  CAS  PubMed  Google Scholar 

  22. Monfoulet L, Malaval L, Aubin JE, Rittling SR, Gadeau AP, Fricain JC, Chassande O (2009) Bone sialoprotein, but not osteopontin, deficiency impairs the mineralization of regenerating bone during cortical defect healing. Bone (in press)

  23. Ayukawa Y, Yasukawa E, Moriyama Y, Ogino Y, Wada H, Atsuta I, Koyano K (2009) Local application of statin promotes bone repair through the suppression of osteoclasts and the enhancement of osteoblasts at bone-healing sites in rats. Oral Surg Oral Med Oral Pathol Oral Radiol Endod 107:336–342

    Article  PubMed  Google Scholar 

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Acknowledgement

We thank Franck Simonnet from the conventional animal facility of Université Bordeaux 2 for technical assistance. This work was supported by INSERM, Région Aquitaine, and Université Bordeaux 2.

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Authors and Affiliations

  1. Université Victor Segalen Bordeaux 2, Bordeaux, 33076, France

    Laurent Monfoulet, Bénédicte Rabier, Olivier Chassande & Jean-Christophe Fricain

  2. INSERM, U 577, Bordeaux, 33076, France

    Laurent Monfoulet, Bénédicte Rabier, Olivier Chassande & Jean-Christophe Fricain

  3. Plateforme Technologique d’Innovation Biomédicale, Université Victor Segalen Bordeaux 2, CHU de, Bordeaux, 33604, France

    Olivier Chassande

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  1. Laurent Monfoulet
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  2. Bénédicte Rabier
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  3. Olivier Chassande
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  4. Jean-Christophe Fricain
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Correspondence to Olivier Chassande.

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The authors have stated that they have no conflicts of interest.

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Monfoulet, L., Rabier, B., Chassande, O. et al. Drilled Hole Defects in Mouse Femur as Models of Intramembranous Cortical and Cancellous Bone Regeneration. Calcif Tissue Int 86, 72–81 (2010). https://doi.org/10.1007/s00223-009-9314-y

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  • DOI: https://doi.org/10.1007/s00223-009-9314-y

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