Method Article

An Efficient and Reproducible Protocol for Distraction Osteogenesis in a Rat Model Leading to a Functional Regenerated Femur

DOI:

10.3791/56433

⸱

October 23rd, 2017

 ,  ,  ,  ,  ,  ,  , 
1CNRS, ISM, Inst Movement Sci, Aix Marseille Univ, 2Sainte-Marguerite Hospital, Institute for Locomotion, Department of Orthopaedics and Traumatology, APHM, 3Sainte-Marguerite Hospital, Institute for Locomotion, Department of Peadiatric Orthopaedics, APHM, 4Ecole centrale de Marseille, 5Faculté de Pharmacie, Laboratoire de Biochimie, 6Service Central de la Qualité et de l'Information Pharmaceutiques, APHM

* These authors contributed equally

In This Article

Summary

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This study describes a reproducible and detailed protocol using a newly developed external fixator for distraction osteogenesis (DO) in a femoral rat model which permits physiological weight-bearing by the animal after removal of the external fixator.

Abstract

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This protocol describes the use of a newly developed external fixator for distraction osteogenesis in a rat femoral model. Distraction osteogenesis (DO) is a surgical technique leading to bone regeneration after an osteotomy. The osteotomized extremities are moved away from each other by gradual distraction to reach the desired elongation. This procedure is widely used in humans for lower and upper limb lengthening, treatment after a bone nonunion, or the regeneration of a bone defect following surgery for bone tumor excision, as well as in maxillofacial reconstruction. Only a few studies clearly demonstrate the efficiency of their protocol in obtaining a functional regenerated bone, i.e., bone that will support physiological weight-bearing without fracture after removal of the external fixator. Moreover, protocols for DO vary and reproducibility is limited by lack of information, making comparison between studies difficult. The aim of this study was to develop a reproducible protocol comprising an appropriate external fixator design for rat limb lengthening, with a detailed surgical technique that permits physiological weight-bearing by the animal after removal of the external fixator.

Introduction

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Distraction osteogenesis (DO) is a surgical technique widely used clinically1,2,3,4 in humans for lower1,2 and upper3 limb lengthening, treatment after a bone nonunion, or the regeneration of a bone defect following surgery for bone tumor excision as well as in maxillofacial reconstruction4. DO leads to bone regeneration after placement of an external fixator in bone and osteotomy. The osteotomized extremities are moved away from each o....

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Protocol

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All procedures described were approved by the University of Aix-Marseille institutional animal care and use committee and the French research ministry and performed in the conventional animal house of Marseille Medical Faculty (France).

1. Define Functional Specifications of the External Fixator Based on the Following Guidelines

  1. Optimize bone anchorage.
    1. Implant half-threaded pins with a diameter (of the threaded section) of 1 mm.
  2. Choose a design to reduce discomfort for the animal.
    1. Choose an external fixator of a small size that fits into a volume ....

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Results

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The X-ray images taken from the end of surgical procedure to the end of consolidation showed no loosening of the half-threaded pins in the femur, indicating stable anchorage. The pins were parallel and well-preserved. The osteotomized extremities were well-aligned along the longitudinal axis of the bone during the DO process (Figure 2). At the end of the latency period, no calcified areas were visible (Figure 2B).......

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Discussion

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This study describes a reproducible protocol comprising an appropriate external fixator design for rat limb lengthening, with a detailed surgical technique that permits physiological weight-bearing by the animal after removal of the external fixator. Our DO protocol led to a functional regenerated bone. After 47 days of consolidation, removal of the homemade external fixator and 2 days of physiological weight-bearing by the animal did not induce any fracture of the regenerating callus. Thanks to the micro-CT reconstructi.......

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Disclosures

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The authors have nothing to disclose.

Acknowledgements

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This work was supported and funded by the CNRS Mecabio challenge.

The authors thank the animal care technician for taking care of the animals throughout the procedure. The authors also acknowledge IVTV central Lyon, through Thierry Hoc. Thanks to Marjorie Sweetko for language revision.

We are greatful to Marylène Lallemand, Cécile Génovésio, and Patrick Laurent for their contribution to this experimental study.

....

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Materials

List of materials used in this article
NameCompanyCatalog NumberComments
KétamineRenaudin578 540-2Supply by animal house
MédétomidineVirbac6799091Supply by animal house
SevofluraneCentravet567 477-2Supply by animal house
BuprenorphineIndivor France3400932731060Supply by animal house
EnrofloxacineChannelPharmaceutical FacturingFR/V/4955220Supply by animal house
PiezotomeSatelec ActeonF57510
Heating pet padTherasageAL8365936Supply by the animal house
Dental X-rayS.A.R.L Innovation médicales et dentairesWYZ - BLUEX
Winiwix SoftwareSoftys DentalPFT
Micro-CT systemnanoScan SPECT/CTGEIT-31105EN (05/14)Subcontract by IVTV central Lyon
Micro-CT analysis Softwarephoenix datos X2 reconstructionnoneFree software
Electric razorBrawnGT415Supply by animal house
Senn’s retractorsWord Precision Instruments501718Blunt version
Betadine SolutionMundipharma Medical CompanyD08AG02Supply by animal house
Resorbable suture thread (5.0)EthiconJV1023Supply by animal house
RugineWord Precision Instruments503406
Mayo-Hegar needle holderWord Precision InstrumentsV503382
Metal drillBeuterlockV020944018003
Micro Olsen-Hegar Needle-holderWord Precision Instruments501989
Mayo scissorWord Precision Instruments501752
ScalpelWord Precision Instruments500236
Sprague-DawleyJanviernone12 weaks and male

References

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  1. Jauregui, J. J., Ventimiglia, A. V., Grieco, P. W., Frumberg, D. B., Herznberg, J. E. Regenerate Bone stimulation following limb lengthening: a meta-analysis. BMC Musculoskelet Disord. 17 (1), 407(2016).
  2. Morcos, M. W., Al-Jallad, H., Hamdy, R.

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Tags

Distraction OsteogenesisRat ModelExternal FixatorFemoral LengtheningBone RegenerationSurgical TechniqueOsteotomy ProcedurePin FixationX ray ImagingMicro CT Analysis

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