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Assessment of subchondral bone microdamage quantification using contrast-enhanced imaging techniques.

Babatunde A Ayodele1, Fatemeh Malekipour2, Charles N Pagel1

  • 1Melbourne Veterinary School, University of Melbourne, Melbourne, Victoria, Australia.

Journal of Anatomy
|March 14, 2024
PubMed
Summary

This study developed a new method to quantify bone microdamage in racehorses using lead uranyl acetate (LUA) staining and advanced imaging. The modified protocol effectively detected microdamage in subchondral bone (SCB), aiding future research on joint health.

Keywords:
back‐scattered scanning electron microscopy (BSEM)bone microdamagedamaged bone fractionmicro‐computed tomographysubchondral bone

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Area of Science:

  • Biomechanical Engineering
  • Veterinary Medicine
  • Materials Science

Background:

  • Subchondral bone (SCB) microdamage is prevalent in athletic animals and humans due to high-impact loading.
  • Microdamage significantly alters bone's biomechanical properties, necessitating accurate quantification methods.
  • Existing methods for microdamage quantification in SCB are limited, especially compared to trabecular bone.

Purpose of the Study:

  • To assess lead uranyl acetate (LUA) staining and contrast-enhanced microcomputed tomography (μCT) for detecting and quantifying SCB microdamage.
  • To determine the specificity of LUA to microdamage and the accuracy of damaged bone volume metrics.
  • To establish a reliable method for quantifying microdamage in SCB, crucial for understanding joint physiology.

Main Methods:

  • SCB specimens from racehorse metacarpus were used, with and without hyaline articular cartilage (HAC).
  • Specimens were subjected to ex vivo uniaxial compression or preserved for in vivo damage.
  • LUA staining (modified protocol) followed by μCT and backscattered scanning electron microscopy (SEM) imaging for quantification.

Main Results:

  • A modified LUA protocol enhanced label penetration into SCB.
  • LUA specifically labeled microdamage, including diffuse, linear, and oblique cracks, excluding those with mineral infills.
  • μCT-based labeled bone volume fraction (LV/BV) was higher without HAC; a corrected metric (LV/BV BS⁻¹) was proposed for accurate quantification.

Conclusions:

  • The combination of HAC removal and a modified LUA staining protocol provides effective staining of damaged SCB.
  • This technique accurately quantifies microdamage in racehorse SCB, offering a valuable tool for research.
  • The developed method facilitates future investigations into the impact of microdamage on joint health and physiology.