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Journal of orthopaedic research : official publication of the Orthopaedic Research Society2024; 42(10); 2264-2276; doi: 10.1002/jor.25869

Longitudinal in vivo cationic contrast-enhanced computed tomography classifies equine articular cartilage injury and repair.

Abstract: Cationic contrast-enhanced computed tomography (CECT) capitalizes on increased contrast agent affinity to the charged proteoglycans in articular cartilage matrix to provide quantitative assessment of proteoglycan content with enhanced images. While high resolution microCT has demonstrated success, we investigate cationic CECT use in longitudinal in vivo imaging at clinical resolution. We hypothesize that repeated administration of CA4+ will have no adverse side effects or complications, and that sequential in vivo imaging assessments will distinguish articular cartilage repair tissue from early degenerative and healthy cartilage in critically sized chondral defects. In an established equine translational preclinical model, lameness and synovial effusion scores are similar to controls after repeated injections of CA4+ (eight injections over 16 weeks) compared to controls. Synovial fluid total protein, leukocyte concentration, and sGAG and PGE concentrations and articular cartilage and synovial membrane scores are also equivalent to controls. Longitudinal in vivo cationic CECT attenuation in repair tissue is significantly lower than peripheral to (adjacent) and distantly from defects (remote sites) by 4 weeks (p < 0.001), and this difference persists until 16 weeks. At the 6- and 8-week time points, the adjacent locations exhibit significantly lower cationic CECT attenuation compared with the remote sites, reflecting peri-defect degeneration (p < 0.01). Cationic CECT attenuation at clinical resolution significantly correlates with cationic CECT (microCT) (r = 0.69, p < 0.0001), sGAG (r = 0.48, p < 0.0001), and ICRS II histology score (r = 0.63, p < 0.0001). In vivo cationic CECT imaging at clinical resolution distinguishes fibrous repair tissue from degenerative and healthy hyaline cartilage and correlates with molecular tissue properties of articular cartilage.
© 2024 The Authors. Journal of Orthopaedic Research® published by Wiley Periodicals LLC on behalf of Orthopaedic Research Society.
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Publication Date: 2024-05-08 PubMed ID: 38715519DOI: 10.1002/jor.25869Google Scholar: Lookup
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Summary

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Overview

  • This study evaluates the use of cationic contrast-enhanced computed tomography (CECT) for non-invasive, longitudinal imaging to differentiate between healthy, degenerative, and repair tissue in equine articular cartilage injuries.
  • It further assesses the safety of repeated contrast agent administration and correlates imaging results with molecular and histological markers of cartilage health.

Background and Purpose

  • Articular cartilage contains proteoglycans, which are negatively charged molecules critical for the tissue’s function and health.
  • Cationic contrast agents such as CA4+ have a positive charge that leads to preferential binding to these proteoglycans, allowing enhanced visualization of cartilage composition using computed tomography (CT).
  • Previous use of microCT with cationic agents has shown promise in detecting cartilage changes ex vivo at very high resolution.
  • The study aims to translate this approach for in vivo longitudinal imaging at clinical CT resolution, which is lower but more applicable for live animals or patients.
  • The hypothesis is twofold: repeated CA4+ injections are safe without causing harm, and in vivo CECT can reliably distinguish repair tissue from degenerative and normal cartilage in equine models of cartilage injury.

Methods

  • Used an established equine (horse) translational preclinical model with critically sized chondral defects (injuries involving cartilage only).
  • Employed repeated intra-articular injections of CA4+ contrast agent — eight injections over 16 weeks.
  • Monitored clinical signs such as lameness and synovial effusion (joint fluid swelling) to assess safety and adverse effects.
  • Collected synovial fluid samples to measure many biochemical markers:
    • Total protein concentration
    • Leukocyte (white blood cell) concentration
    • Sulfated glycosaminoglycan (sGAG) concentration — a key component of cartilage extracellular matrix
    • Prostaglandin E (PGE) concentration — related to inflammation
  • Histological scoring of articular cartilage and synovial membrane was performed post-mortem.
  • Longitudinal in vivo CECT imaging was conducted at multiple time points over 16 weeks and compared regions:
    • Repair tissue inside the defect
    • Adjacent cartilage surrounding the defect (peri-defect)
    • Remote cartilage distant from the defect
  • Correlated clinical resolution CECT attenuation with microCT results, biochemical sGAG content, and histological grading (ICRS II score).

Key Findings

  • Safety Assessment:
    • No significant differences in lameness or synovial effusion scores between treated and control joints.
    • Synovial fluid biomarkers (protein, leukocytes, sGAG, PGE) showed no adverse inflammatory or degradative changes due to repeated CA4+ administration.
    • Histological evaluation showed cartilage and synovial membrane integrity was maintained, supporting safety.
  • Imaging Results:
    • CECT attenuation in repair tissue inside defects was significantly lower than adjacent and remote areas starting as early as 4 weeks post-injury, persisting through 16 weeks.
    • Adjacent cartilage had lower attenuation than remote cartilage at 6 and 8 weeks, indicating early degenerative changes around the injury site (peri-defect degeneration).
  • Correlations and Validation:
    • CECT attenuation at clinical CT resolution correlated strongly with microCT (r=0.69), validating the imaging technique.
    • Positive but moderate correlations were found between CECT signal and biochemical sGAG content (r=0.48), confirming that imaging reflects molecular tissue composition.
    • Histology scores related to cartilage quality also correlated well with CECT attenuation (r=0.63), supporting clinical relevance.
  • Interpretation:
    • Lower attenuation in repair tissue reflects reduced proteoglycan content and fibrous rather than healthy hyaline cartilage formation.
    • Identifying peri-defect degeneration early may help guide clinical interventions.
    • CECT provides a non-invasive way to monitor cartilage repair quality and disease progression longitudinally.

Implications and Conclusion

  • The study demonstrates that repeated intra-articular injections of a cationic contrast agent in horses are safe and do not provoke adverse joint inflammation.
  • Longitudinal in vivo CECT imaging at clinical CT resolution can successfully differentiate repair tissue, degenerating cartilage, and healthy cartilage based on proteoglycan content.
  • This methodology allows for quantitative, non-destructive, and repeated monitoring of cartilage repair in a clinically relevant animal model.
  • Future applications may include improved diagnostics and treatment monitoring for joint injuries and osteoarthritis in both veterinary and human medicine.

Cite This Article

APA
Nelson BB, Mäkelä JTA, Lawson TB, Patwa AN, Snyder BD, McIlwraith CW, Grinstaff MW, Seabaugh KA, Barrett MF, Goodrich LR, Kawcak CE. (2024). Longitudinal in vivo cationic contrast-enhanced computed tomography classifies equine articular cartilage injury and repair. J Orthop Res, 42(10), 2264-2276. https://doi.org/10.1002/jor.25869

Publication

Journal of orthopaedic research : official publication of the Orthopaedic Research Society
ISSN: 1554-527X
NlmUniqueID: 8404726
Country: United States
Language: English
Volume: 42
Issue: 10
Pages: 2264-2276

Researcher Affiliations

Nelson, Brad B
  • Orthopaedic Research Center, C. Wayne McIlwraith Translational Medicine Institute, Colorado State University, Fort Collins, Colorado, USA.
Mäkelä, Janne T A
  • Harvard Medical School, Beth Israel Deaconess Medical Center, Center for Advanced Orthopaedic Studies, Boston, Massachusetts, USA.
  • Department of Technical Physics, University of Eastern Finland, Kuopio, Finland.
  • Departments of Chemistry and Biomedical Engineering, Boston University, Boston, Massachusetts, USA.
Lawson, Taylor B
  • Harvard Medical School, Beth Israel Deaconess Medical Center, Center for Advanced Orthopaedic Studies, Boston, Massachusetts, USA.
  • Departments of Chemistry and Biomedical Engineering, Boston University, Boston, Massachusetts, USA.
Patwa, Amit N
  • Departments of Chemistry and Biomedical Engineering, Boston University, Boston, Massachusetts, USA.
  • Deparment of Chemistry, School of Science, Navrachana University, Vadodara, Gujarat, India.
Snyder, Brian D
  • Harvard Medical School, Beth Israel Deaconess Medical Center, Center for Advanced Orthopaedic Studies, Boston, Massachusetts, USA.
McIlwraith, C Wayne
  • Orthopaedic Research Center, C. Wayne McIlwraith Translational Medicine Institute, Colorado State University, Fort Collins, Colorado, USA.
Grinstaff, Mark W
  • Departments of Chemistry and Biomedical Engineering, Boston University, Boston, Massachusetts, USA.
Seabaugh, Kathryn A
  • Orthopaedic Research Center, C. Wayne McIlwraith Translational Medicine Institute, Colorado State University, Fort Collins, Colorado, USA.
Barrett, Myra F
  • Orthopaedic Research Center, C. Wayne McIlwraith Translational Medicine Institute, Colorado State University, Fort Collins, Colorado, USA.
Goodrich, Laurie R
  • Orthopaedic Research Center, C. Wayne McIlwraith Translational Medicine Institute, Colorado State University, Fort Collins, Colorado, USA.
Kawcak, Christopher E
  • Orthopaedic Research Center, C. Wayne McIlwraith Translational Medicine Institute, Colorado State University, Fort Collins, Colorado, USA.

MeSH Terms

  • Animals
  • Horses
  • Cartilage, Articular / diagnostic imaging
  • Cartilage, Articular / injuries
  • Contrast Media
  • Tomography, X-Ray Computed
  • Female
  • Cations
  • Male

Grant Funding

  • Päivikki ja Sakari Sohlbergin Säätiö
  • Suomen Kulttuurirahasto
  • Grayson-Jockey Club Research Foundation
  • College of Veterinary Medicine and Biomedical Sciences Cooperative Veterinary Scientist Research Training Fellowship, Colorado State University
  • William Fairfield Warren Distinguished Professorship
  • Orion Research Foundation
  • Finnish Cultural Foundation

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