Skip to main content

The Murine Femoral Bone Graft Model and a Semiautomated Histomorphometric Analysis Tool

  • Protocol
Skeletal Development and Repair

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1130))

Abstract

Preclinical studies of bone repair remain a high priority because of unresolved clinical problems associated with treating critical segmental defects and complications of fracture healing. Over the last decade, the murine femoral allograft model has gained popularity due to its standardized surgery and potential for examining a vast array of radiographic, biomechanical, and histological outcome measures. Here, we describe these methods and a novel semiautomated histomorphometric approach to quantify the amount of bone, cartilage, and undifferentiated mesenchymal tissue in demineralized paraffin sections of allografted murine femurs using the Visiopharm Image Analysis Software System.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 84.99
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 139.00
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 109.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

References

  1. Xie C, Reynolds D, Awad H, Rubery PT, Pelled G, Gazit D, Guldberg RE, Schwarz EM, O'Keefe RJ, Zhang X (2007) Structural bone allograft combined with genetically engineered mesenchymal stem cells as a novel platform for bone tissue engineering. Tissue Eng 13(3):435–445 [Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov't]

    Article  CAS  PubMed  Google Scholar 

  2. Xie C, Xue M, Wang Q, Schwarz EM, O'Keefe RJ, Zhang X (2008) Tamoxifen-inducible CreER-mediated gene targeting in periosteum via bone-graft transplantation. J Bone Joint Surg Am 90(1):9–13 [Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov't]

    Article  PubMed  PubMed Central  Google Scholar 

  3. Xie C, Liang B, Xue M, Lin AS, Loiselle A, Schwarz EM, Guldberg RE, O'Keefe RJ, Zhang X (2009) Rescue of impaired fracture healing in COX-2-/- mice via activation of prostaglandin E2 receptor subtype 4. Am J Pathol 175(2):772–785 [Research Support, N.I.H., Extramural]

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  4. Dhillon RS, Xie C, Tyler W, Calvi LM, Awad HA, Zuscik MJ, O'Keefe RJ, Schwarz EM (2012) PTH enhanced structural allograft healing is associated with decreased angiopoietin-2 mediated arteriogenesis, mast cell accumulation and fibrosis. J Bone Miner Res 18:1–34

    Google Scholar 

  5. Zhang X, Xie C, Lin AS, Ito H, Awad H, Lieberman JR, Rubery PT, Schwarz EM, O'Keefe RJ, Guldberg RE (2005) Periosteal progenitor cell fate in segmental cortical bone graft transplantations: implications for functional tissue engineering. J Bone Miner Res 20(12):2124–2137

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  6. Naik AA, Xie C, Zuscik MJ, Kingsley P, Schwarz EM, Awad H, Guldberg R, Drissi H, Puzas JE, Boyce B, Zhang X, O'Keefe RJ (2009) Reduced COX-2 expression in aged mice is associated with impaired fracture healing. J Bone Miner Res 24(2):251–264 [Research Support, N.I.H., Extramural]

    Article  CAS  PubMed  Google Scholar 

  7. Tiyapatanaputi P, Rubery PT, Carmouche J, Schwarz EM, O'Keefe RJ, Zhang X (2004) A novel murine segmental femoral graft model. J Orthop Res 22(6):1254–1260

    Article  PubMed  Google Scholar 

  8. Xie C, Ming X, Wang Q, Schwarz EM, Guldberg RE, O'Keefe RJ, Zhang X (2008) COX-2 from the injury milieu is critical for the initiation of periosteal progenitor cell mediated bone healing. Bone 43(6):1075–1083

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  9. Yazici C, Takahata M, Reynolds DG, Xie C, Samulski RJ, Samulski J, Beecham EJ, Gertzman AA, Spilker M, Zhang X, O'Keefe RJ, Awad HA, Schwarz EM (2011) Self-complementary AAV2.5-BMP2-coated femoral allografts mediated superior bone healing versus live autografts in mice with equivalent biomechanics to unfractured femur. Mol Ther 19(8):1416–1425

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  10. Dhillon RS, Xie C, Tyler W, Calvi LM, Awad HA, Zuscik MJ, O'Keefe RJ, Schwarz EM (2012) PTH-enhanced structural allograft healing is associated with decreased angiopoietin-2-mediated arteriogenesis, mast cell accumulation, and fibrosis. J Bone Miner Res 28(3):586–597

    Article  Google Scholar 

  11. Program NLAS (2012) Ketamine-xylazine combination for rodent anesthesia. New York: NIH. http://ncifrederick.cancer.gov/rtp/lasp/intra/acuc/beth/KetamineXylazine.asp. Accessed 13 Nov 2012

    Google Scholar 

  12. Pritchett-Corning KR, Luo Y, Mulder GB, White WJ (2011) Principles of rodent surgery for the new surgeon. J Visual Exp 2011(47):[Video-Audio Media]

    Google Scholar 

  13. Zhang X, Schwarz EM, Young DA, Puzas JE, Rosier RN, O'Keefe RJ (2002) Cyclo-oxygenase-2 regulates mesenchymal cell differentiation into the osteoblast lineage and is critically involved in bone repair. J Clin Invest 109(11):1405–1415 [Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.]

    Article  CAS  PubMed  PubMed Central  Google Scholar 

Download references

Acknowledgements

The authors would like to thank Ryan Tierney, Sarah Mack, Kathleen Maltby, and Ashish Thomas for their assistance with the histology and whole-slide scanning. This work was supported by research grants from the National Institutes of Health (1S10RR027340, DE19902, AR54041, and P30 AR061307).

Author information

Authors and Affiliations

Authors

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

© 2014 Springer Science+Business Media, LLC

About this protocol

Cite this protocol

Dhillon, R.S., Zhang, L., Schwarz, E.M., Boyce, B.F., Xie, C. (2014). The Murine Femoral Bone Graft Model and a Semiautomated Histomorphometric Analysis Tool. In: Hilton, M. (eds) Skeletal Development and Repair. Methods in Molecular Biology, vol 1130. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-989-5_4

Download citation

  • DOI: https://doi.org/10.1007/978-1-62703-989-5_4

  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-62703-988-8

  • Online ISBN: 978-1-62703-989-5

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics