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Home / Equine Research Bank / Osteoarthritis Cartilage / Identification of cartilage injury using quantitative multip...
Osteoarthritis and cartilage2013; 22(2); 355-362; doi: 10.1016/j.joca.2013.10.008

Identification of cartilage injury using quantitative multiphoton microscopy.

Abstract: Cartilage injury can lead to post-traumatic osteoarthritis (PTOA). Immediate post-trauma cellular and structural changes are not widely understood. Furthermore, current cellular-resolution cartilage imaging techniques require sectioning of cartilage and/or use of dyes not suitable for patient imaging. In this study, we used multiphoton microscopy (MPM) data with FDA-approved sodium fluorescein to identify and evaluate the pattern of chondrocyte death after traumatic injury. Methods: Mature equine distal metacarpal or metatarsal osteochondral blocks (OCBs) were injured by 30 MPa compressive loading delivered over 1 s. Injured and control sites were imaged unfixed and in situ 1 h post-injury with sodium fluorescein using rasterized z-scanning. MPM data was quantified in MATLAB, reconstructed in 3-D, and projected in 2-D to determine the damage pattern. Results: MPM images (600 per sample) were reconstructed and analyzed for cell death. The overall distribution of cell death appeared to cluster into circular (n = 7) or elliptical (n = 4) patterns (p = 0.006). Dead cells were prevalent near cracks in the matrix, with only 26.3% (SE = 5.0%, p < 0.0001) of chondrocytes near cracks being viable. Conclusions: This study demonstrates the first application of MPM for evaluating cellular-scale cartilage injury in situ in live tissue, with clinical potential for detecting early cartilage damage. With this technique, we were able to uniquely observe two death patterns resulting from the same compressive loading, which may be related to local variability in matrix structure. These results also demonstrate proof-of-concept MPM diagnostic use in detecting subtle and early cartilage damage not detectable in any other way.
Copyright © 2013 Osteoarthritis Research Society International. Published by Elsevier Ltd. All rights reserved.
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Publication Date: 2013-11-01 PubMed ID: 24185113PubMed Central: PMC4117377DOI: 10.1016/j.joca.2013.10.008Google Scholar: Lookup
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  • Evaluation Study
  • Journal Article
  • Research Support
  • N.I.H.
  • Extramural
  • Research Support
  • Non-U.S. Gov't

Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

This research focuses on the use of multiphoton microscopy to identify cell death patterns in injured cartilage tissue, a development critical to understanding and diagnosing early cartilage damage that can lead to conditions such as post-traumatic osteoarthritis.

Study Methodology

  • The researchers used osteochondral blocks (cartilage joined to bone) from mature horses.
  • These blocks were subjected to compressive loading of 30 MPa for 1 second to simulate injury.
  • An hour after the injury was inflicted, the blocks were scanned with FDA-approved sodium fluorescein and multiphoton microscopy in their native, unaltered state.
  • The data from the microscopy was quantified and used to create both 3-dimensional reconstructions and 2-dimensional projections of the cartilage to evaluate the pattern of cell death.

Results and Findings

  • 600 images per sample were reconstructed and evaluated for cartilage cell death.
  • Two distinct patterns of cartilage cell death were observed: circular and elliptical.
  • Dead cells were frequently found near matrix cracks, with only roughly 26% of cells near these flaws remaining alive.
  • The patterns of cell death may be linked to local variations in the structure of the cartilage matrix.

Implications and Conclusions

  • This research marks the first time multiphoton microscopy has been used to evaluate cartilage injury at the cellular level, providing a promising potential clinical tool for early cartilage damage detection. It’s a technique that could provide valuable insight into the immediate aftermath of a traumatic cartilage injury, something not adequately understood at the moment.
  • The detected patterns of cell death may provide valuable insights into cartilage injury physiology, which could ultimately lead to improved diagnostic and treatment strategies for cartilage injury and related conditions.
  • The results also demonstrate that multiphoton microscopy could potentially be used as a diagnostic tool for detecting early and subtle cartilage damage that isn’t identifiable via other methods.

Cite This Article

APA
Novakofski KD, Williams RM, Fortier LA, Mohammed HO, Zipfel WR, Bonassar LJ. (2013). Identification of cartilage injury using quantitative multiphoton microscopy. Osteoarthritis Cartilage, 22(2), 355-362. https://doi.org/10.1016/j.joca.2013.10.008

Publication

Osteoarthritis and cartilage
ISSN: 1522-9653
NlmUniqueID: 9305697
Country: England
Language: English
Volume: 22
Issue: 2
Pages: 355-362
PII: S1063-4584(13)00981-3

Researcher Affiliations

Novakofski, K D
  • Department of Clinical Sciences, Cornell University, Ithaca, NY, USA.
Williams, R M
  • Department of Biomedical Engineering, Cornell University, Ithaca, NY 14853, USA.
Fortier, L A
  • Department of Clinical Sciences, Cornell University, Ithaca, NY, USA.
Mohammed, H O
  • Population Medicine and Diagnostic Sciences, Cornell University, Ithaca, NY 14853, USA.
Zipfel, W R
  • Department of Biomedical Engineering, Cornell University, Ithaca, NY 14853, USA.
Bonassar, L J
  • Department of Biomedical Engineering, Cornell University, Ithaca, NY 14853, USA. Electronic address: lb244@cornell.edu.

MeSH Terms

  • Animals
  • Cartilage, Articular / injuries
  • Cartilage, Articular / pathology
  • Cell Death / physiology
  • Chondrocytes / pathology
  • Disease Models, Animal
  • Early Diagnosis
  • Feasibility Studies
  • Horses
  • Image Processing, Computer-Assisted / methods
  • Metacarpus / injuries
  • Metacarpus / pathology
  • Metatarsal Bones / injuries
  • Metatarsal Bones / pathology
  • Microscopy, Fluorescence, Multiphoton / methods
  • Stress, Mechanical
  • Weight-Bearing

Grant Funding

  • TL1 TR000459 / NCATS NIH HHS
  • UL1 TR000457 / NCATS NIH HHS
  • TL1RR000459 / NCRR NIH HHS
  • P41-EB01976 / NIBIB NIH HHS

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Citations

This article has been cited 7 times.
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