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Quantifying Massive Allograft Healing of the Canine Femur In Vivo and Ex Vivo: A Pilot Study

  • Symposium: Allograft Research and Transplantation
  • Published:
Clinical Orthopaedics and Related Research®

Abstract

Background

Allograft integration in segmental osseous defects is unpredictable. Imaging techniques have not been applied to investigate angiogenesis and bone formation during allograft healing in a large-animal model.

Questions/purposes

We used dynamic contrast-enhanced (DCE)-MRI and cone beam (CB)-CT to quantify vascularity and bone volume in a canine femoral allograft model and determined their relationship with biomechanical testing and histomorphometry.

Methods

Femoral ostectomy was performed in three dogs and reconstructed with a 5-cm allograft and compression plate. At 0.5, 3, and 6 months, we performed DCE-MRI to quantify vascular permeability (Ktrans) and perfused fraction and CB-CT to quantify bone volume. We also performed posteuthanasia torsional testing and dynamic histomorphometry of the grafted and nonoperated femurs.

Results

DCE-MRI confirmed the avascular nature of allograft healing (perfused fraction, 2.08%–3.25%). CB-CT demonstrated new bone formation at 3 months (26.2, 3.7, and 2.2 cm3) at the graft-host junctions, which remodeled down at 6 months (14.0, 2.2, and 2.0 cm3). The increased bone volume in one subject was confirmed with elevated Ktrans (0.22) at 3 months. CB-CT-identified remodeled bone at 6 months was corroborated by histomorphometry. Allografted femurs recovered only 40% of their strength at 6 months.

Conclusions

CB-CT and DCE-MRI can discriminate differences in angiogenesis and bone formation in the canine allograft model, which has potential to detect a small (32%) drug or device effect on biomechanical healing with only five animals per group.

Clinical Relevance

These radiographic tools may have the potential to assess adjuvant effects on vascular invasion and new bone formation after segmental allograft transplantation.

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Acknowledgments

We thank our collaborative authors from the University of Rochester, Masahiko Takahata, MD, and Chao Xie, MD, for their assistance in the daily observations of the canine subjects and care for the animals during DCE-MRI and CB-CT; David Conover, MS, for his help running the CB-CT scanner; and Hani A. Awad, PhD, for help with the experimental design and data analyses. We also thank Synthes Inc for providing surgical equipment and implants.

Author information

Authors and Affiliations

  1. Phillip Spiegel Orthopaedic Research Laboratory, Foundation for Orthopaedic Research and Education, Tampa, FL, USA

    Brandon G. Santoni PhD

  2. Department of Clinical Sciences, Colorado State University, Fort Collins, CO, USA

    Nicole Ehrhart VMD, MS

  3. Department of Imaging Sciences, University of Rochester, Rochester, NY, USA

    Ricardo Betancourt-Benitez PhD

  4. Department of Biostatistics & Computational Biology, University of Rochester, Rochester, NY, USA

    Christopher A. Beck PhD

  5. Department of Biomedical Engineering, University of Rochester, Rochester, NY, USA

    Edward M. Schwarz PhD

  6. The Center for Musculoskeletal Research, University of Rochester Medical Center, 601 Elmwood Avenue, Box 665, Rochester, NY, 14642, USA

    Edward M. Schwarz PhD

Authors
  1. Brandon G. Santoni PhD
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  2. Nicole Ehrhart VMD, MS
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  3. Ricardo Betancourt-Benitez PhD
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  4. Christopher A. Beck PhD
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  5. Edward M. Schwarz PhD
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Corresponding author

Correspondence to Edward M. Schwarz PhD.

Additional information

One or more of the authors (BGS, NE, EMS) have received funding from the Musculoskeletal Transplant Foundation (Edison, NJ, USA). One or more of the authors have received funding from the following grants: NIH PHS AR054041 (EMS), AR056696 (EMS), DE019902 (EMS), AR061307 (EMS), and 1R01EB012048-01A1 (BGS).

All ICMJE Conflict of Interest Forms for authors and Clinical Orthopaedics and Related Research editors and board members are on file with the publication and can be viewed on request.

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

All in vivo work and ex vivo cone beam CT and imaging analyses were performed at the University of Rochester, Rochester, NY, USA. The ex vivo biomechanics, histology, and histomorphometry were performed at Colorado State University, Fort Collins, CO, USA.

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Santoni, B.G., Ehrhart, N., Betancourt-Benitez, R. et al. Quantifying Massive Allograft Healing of the Canine Femur In Vivo and Ex Vivo: A Pilot Study. Clin Orthop Relat Res 470, 2478–2487 (2012). https://doi.org/10.1007/s11999-012-2349-9

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  • DOI: https://doi.org/10.1007/s11999-012-2349-9

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