Research Article
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Published Online: 3 May 2021

High Frequency Spectral Ultrasound Imaging Detects Early Heterotopic Ossification in Rodents

Publication: Stem Cells and Development
Volume 30, Issue Number 9

Abstract

Heterotopic ossification (HO) is a devastating condition in which ectopic bone forms inappropriately in soft tissues following traumatic injuries and orthopedic surgeries as a result of aberrant mesenchymal progenitor cell (MPC) differentiation. HO leads to chronic pain, decreased range of motion, and an overall decrease in quality of life. While several treatments have shown promise in animal models, all must be given during early stages of formation. Methods for early determination of whether and where endochondral ossification/soft tissue mineralization (HO anlagen) develop are lacking. At-risk patients are not identified sufficiently early in the process of MPC differentiation and soft tissue endochondral ossification for potential treatments to be effective. Hence, a critical need exists to develop technologies capable of detecting HO anlagen soon after trauma, when treatments are most effective. In this study, we investigate high frequency spectral ultrasound imaging (SUSI) as a noninvasive strategy to identify HO anlagen at early time points after injury. We show that by determining quantitative parameters based on tissue organization and structure, SUSI identifies HO anlagen as early as 1-week postinjury in a mouse model of burn/tenotomy and 3 days postinjury in a rat model of blast/amputation. We analyze single cell RNA sequencing profiles of the MPCs responsible for HO formation and show that the early tissue changes detected by SUSI match chondrogenic and osteogenic gene expression in this population. SUSI identifies sites of soft tissue endochondral ossification at early stages of HO formation so that effective intervention can be targeted when and where it is needed following trauma-induced injury. Furthermore, we characterize the chondrogenic to osteogenic transition that occurs in the MPCs during HO formation and correlate gene expression to SUSI detection of the HO anlagen.

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The contents of this publication are the sole responsibility of the author(s) and do not necessarily reflect the views, opinions, or policies of Uniformed Services University of the Health Sciences (USUHS), The Henry M. Jackson Foundation for the Advancement of Military Medicine, Inc., the Department of Defense (DoD), and the Departments of the Army, Navy, or Air Force. Mention of trade names, commercial products, or organizations does not imply endorsement by the U.S. Government.

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Information & Authors

Information

Published In

cover image Stem Cells and Development
Stem Cells and Development
Volume 30Issue Number 9May 1, 2021
Pages: 473 - 484
PubMed: 33715398

History

Published in print: May 1, 2021
Published online: 3 May 2021
Published ahead of print: 19 April 2021
Published ahead of production: 15 March 2021
Accepted: 12 March 2021
Received: 14 January 2021

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    Authors

    Affiliations

    Nicole J. Edwards
    Department of Surgery, University of Michigan, Ann Arbor, Michigan, USA.
    Eric Hobson
    Department of Biomedical Engineering, University of Michigan, Ann Arbor, Michigan, USA.
    Devaveena Dey
    Department of Surgery, Uniformed Services University of the Health Sciences, Bethesda, Maryland, USA.
    Henry M. Jackson Foundation for the Advancement of Military Medicine, Inc., Bethesda, Maryland, USA.
    Alisha Rhodes
    Department of Surgery, Uniformed Services University of the Health Sciences, Bethesda, Maryland, USA.
    Henry M. Jackson Foundation for the Advancement of Military Medicine, Inc., Bethesda, Maryland, USA.
    Archie Overmann
    Department of Surgery, Uniformed Services University of the Health Sciences, Bethesda, Maryland, USA.
    Benjamin Hoyt
    Department of Surgery, Uniformed Services University of the Health Sciences, Bethesda, Maryland, USA.
    Sarah A. Walsh
    Department of Surgery, Uniformed Services University of the Health Sciences, Bethesda, Maryland, USA.
    Chase A. Pagani
    Department of Surgery, University of Texas Southwestern Medical Center, Dallas, Texas, USA.
    Amy L. Strong
    Department of Surgery, University of Michigan, Ann Arbor, Michigan, USA.
    Geoffrey E. Hespe
    Department of Surgery, University of Michigan, Ann Arbor, Michigan, USA.
    Karthik R. Padmanabhan
    Epigenomic Core, University of Michigan, Ann Arbor, Michigan, USA.
    Amanda Huber
    Department of Surgery, University of Michigan, Ann Arbor, Michigan, USA.
    Cheri Deng
    Department of Biomedical Engineering, University of Michigan, Ann Arbor, Michigan, USA.
    Thomas A. Davis
    Department of Surgery, Uniformed Services University of the Health Sciences, Bethesda, Maryland, USA.
    Benjamin Levi [email protected]
    Department of Surgery, University of Texas Southwestern Medical Center, Dallas, Texas, USA.

    Notes

    Address correspondence to: Dr. Benjamin Levi, Department of Surgery, Center for Organogenesis and Trauma, University of Texas Southwestern Medical Center, 5323 Harry Hines Blvd, Dallas, TX 75390-9158, USA [email protected]

    Author Disclosure Statement

    The authors have nothing to disclose and declare no relevant conflicts of interest.

    Funding Information

    N.J.E. was supported by a Ruth L. Kirschstein Institutional National Research Service Award Postdoctoral fellowship (T32-HD007505). A.L.S. was supported by Ruth L. Kirschstein Institutional National Research Service Award Postdoctoral fellowship (T32-CA009672). GEH was supported by Plastic Surgery Foundation Research Fellowship Grant. B.L. was supported or partially supported by National Institutes of Health (NIH) R01GM123069, R01AR071379, American College of Surgeons Clowes Award, and US Department of Defense Grant W81XWH-18-1-0653 (OR170174). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the article.

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