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Microcomputed tomography–based structural analysis of various bone tissue regeneration models

Nature Protocols volume 6, pages 105–110 (2011)Cite this article

Abstract

Microcomputed tomography (microCT) analysis is a powerful tool for the evaluation of bone tissue because it provides access to the 3D microarchitecture of the bone. It is invaluable for regenerative medicine as it provides the researcher with the opportunity to explore the skeletal system both in vivo and ex vivo. The quantitative assessment of macrostructural characteristics and microstructural features may improve our ability to estimate the quality of newly formed bone. We have developed a unique procedure for analyzing data from microCT scans to evaluate bone structure and repair. This protocol describes the procedures for microCT analysis of three main types of mouse bone regeneration models (ectopic administration of bone-forming mesenchymal stem cells, and administration of cells after both long bone defects and cranial segmental bone defects) that can be easily adapted for a variety of other models. Precise protocols are crucial because the system is extremely user sensitive and results can be easily biased if standardized methods are not applied. The suggested protocol takes 1.5–3.5 h per sample, depending on bone tissue sample size, the type of equipment used, variables of the scanning protocol and the operator's experience.

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Figure 1: Ectopic bone formation model.
Figure 2: Segmental defect in a long bone model.
Figure 3: Round critical-size bone defect model.
Figure 4: Setting the lower threshold.

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Acknowledgements

We acknowledge funding from the National Institutes of Health grant nos. R01AR056694 and R01DE019902 and from the Telemedicine and Advanced Technology Research Center (TATRC), U.S. Army Medical Research and Materiel Command No. 08217008 (W.T., Z.G., G.P. and D.G.).

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Authors and Affiliations

  1. Skeletal Biotech Laboratory, The Hebrew University–Hadassah Faculty of Dental Medicine, Ein Kerem, Jerusalem, Israel

    Ilan Kallai, Olga Mizrahi, Zulma Gazit, Gadi Pelled & Dan Gazit

  2. Department of Surgery and Cedars-Sinai Regenerative Medicine Institute, Cedars-Sinai Medical Center, Los Angeles, California, USA

    Wafa Tawackoli, Zulma Gazit, Gadi Pelled & Dan Gazit

Authors
  1. Ilan Kallai
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  2. Olga Mizrahi
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  3. Wafa Tawackoli
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  4. Zulma Gazit
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  5. Gadi Pelled
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  6. Dan Gazit
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Contributions

I.K. developed the concept, designed the protocol and contributed to writing the manuscript. O.M. provided helpful comments during the development of the protocol and contributed to writing the manuscript. W.T. provided technical support and conceptual advice. G.P. provided helpful comments and contributed to writing the manuscript. Z.G. and D.G. supervised the project. All authors discussed the implications of the protocol and commented on the manuscript at all stages.

Corresponding author

Correspondence to Dan Gazit.

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The authors declare no competing financial interests.

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Kallai, I., Mizrahi, O., Tawackoli, W. et al. Microcomputed tomography–based structural analysis of various bone tissue regeneration models. Nat Protoc 6, 105–110 (2011). https://doi.org/10.1038/nprot.2010.180

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