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Reproducibility and Radiation Effect of High-Resolution In Vivo Micro Computed Tomography Imaging of the Mouse Lumbar Vertebra and Long Bone

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Abstract

A moderate radiation dose, in vivo µCT scanning protocol was developed and validated for long-term monitoring of multiple skeletal sites (femur, tibia, vertebra) in mice. A customized, 3D printed mouse holder was designed and utilized to minimize error associated with animal repositioning, resulting in good to excellent reproducibility in most cortical and trabecular bone microarchitecture and density parameters except for connectivity density. Repeated in vivo µCT scans of mice were performed at the right distal femur and the 4th lumbar vertebra every 3 weeks until euthanized at 9 weeks after the baseline scan. Comparing to the non-radiated counterparts, no radiation effect was found on trabecular bone volume fraction, osteoblast and osteoblast number/surface, or bone formation rate at any skeletal site. However, trabecular number, thickness, and separation, and structure model index were sensitive to ionizing radiation associated with the µCT scans, resulting in subtle but significant changes over multiple scans. Although the extent of radiation damage on most trabecular bone microarchitecture measures are comparable or far less than the age-related changes during the monitoring period, additional considerations need to be taken to minimize the confounding radiation factors when designing experiments using in vivo µCT imaging for long-term monitoring of mouse bone.

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Acknowledgements

Research reported in this publication was supported by the Penn Center for Musculoskeletal Diseases (PCMD) NIH/NIAMS P30-AR069619, NIH/NIAMS K01-AR066743 (to XSL), NIH/NIAMS R01-AR071718 (to XSL), and NSF Graduate Research Fellowship (to CMJdB). We would like to thank Carlos Osuna for constructive criticism of the manuscript.

Conflict of interest

All authors have no conflict of interest.

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Authors and Affiliations

  1. Key Laboratory of Biorheological Science and Technology, Ministry of Education, and Bioengineering College, Chongqing University, Chongqing, China

    Hongbo Zhao & Linhong Deng

  2. McKay Orthopaedic Research Laboratory, Department of Orthopaedic Surgery, Perelman School of Medicine, University of Pennsylvania, 332A Stemmler Hall, 36th Street and Hamilton Walk, Philadelphia, PA, 19104, USA

    Hongbo Zhao, Chih-Chiang Chang, Yang Liu, Youwen Yang, Wei-Ju Tseng, Chantal M. de Bakker, Rebecca Chung, Priyanka Ghosh & X. Sherry Liu

  3. Department of Orthodontics, Stomatological Hospital of Chongqing Medical University, Chongqing, China

    Yang Liu

  4. Changzhou Key Laboratory of Respiratory Medical Engineering, Institute of Biomedical Engineering and Health Sciences, Changzhou University, Changzhou, 213164, Jiangsu, China

    Linhong Deng

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  1. Hongbo Zhao
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Correspondence to Linhong Deng or X. Sherry Liu.

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Electronic supplementary material

A customized holder fixture for in vivo mouse scanning is available for download as a .stl file for 3D printing: For use, the anesthetized mouse is elongated and laid in a supine position along the bed of the holder fixture. The leg of interest is carefully extruded through the hole in the center of the smaller disk. Then, a 6 inch strip of Coban selfadherent wrap (3M, St. Paul, Minnesota, United States) is cut in half longitudinally, stretched, and then wrapped along the leg until it is comfortably secured in the leg holder, while ensuring that the ankle of the mouse is extended beyond the leg holder by approximately 5mm. The vertebra is fixated by wrapping a 6 inch strip of Coban selfadherent wrap around the distal portion of the mouse body (including the tail) and the bed of the holder fixture three times while ensuring that the binding does not hinder breathing. Once the mouse is restrained appropriately, the scanner door may be shut to load the 2D scout view, followed by selecting the regions of interest for in vivo scanning.

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Zhao, H., Chang, CC., Liu, Y. et al. Reproducibility and Radiation Effect of High-Resolution In Vivo Micro Computed Tomography Imaging of the Mouse Lumbar Vertebra and Long Bone. Ann Biomed Eng 48, 157–168 (2020). https://doi.org/10.1007/s10439-019-02323-z

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  • DOI: https://doi.org/10.1007/s10439-019-02323-z

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