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Home / Equine Research Bank / Aging Cell / Immunolabelling and Micro-Computed Tomography Revealed Age-R...
Aging cell2025; 25(1); e70293; doi: 10.1111/acel.70293

Immunolabelling and Micro-Computed Tomography Revealed Age-Related Alterations in 3D Microvasculature of Tendons.

Abstract: Tendon degeneration is common, and its risk increases with age both in humans and horses. Tendon regeneration and healing is limited due to inherent low cell density and vascularisation, and current treatments are insufficient as indicated by scar tissue formation and a high re-injury rate. The tendon vasculature plays a crucial role in tendon homeostasis, regeneration and healing, making it a potential therapeutic target. However, the effect of ageing on the tendon microvasculature is poorly understood. Here, we provide the first comprehensive characterisation of the tendon microvasculature. We employed high-resolution 3D imaging techniques, using micro-computed tomography (μCT) and confocal microscopy, to investigate age-related alterations in the vasculature within the equine superficial digital flexor tendon (SDFT), a functional equivalent of the human Achilles tendon. μCT analysis revealed a well-developed vascular network within the interfascicular matrix (IFM) and demonstrated significant age-associated reductions in vascular volume (70%), vessel diameter (30%) and density (74%). 3D immunolabelling showed significant reductions in MYH11- (96%) and desmin-positive (78%) volumes; however, there was a pronounced age-associated increase in von Willebrand factor (VWF)-positive volume (220%), which was accompanied by a significantly higher (249%) pericyte density. Taken together, these results indicate a loss of larger blood vessels in the IFM but an increase in small vessel formation, suggesting that neo-angiogenesis is induced in aged tendon alongside a loss of vascular homeostasis. These insights enhance our understanding of tendon ageing and may contribute to developing new therapeutic approaches for improving tendon health and repair in older individuals.
© 2025 The Author(s). Aging Cell published by Anatomical Society and John Wiley & Sons Ltd.
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Publication Date: 2025-11-18 PubMed ID: 41250917PubMed Central: PMC12740099DOI: 10.1111/acel.70293Google Scholar: Lookup
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  • Journal Article

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.

Overview

  • This study investigates how aging affects the tiny blood vessels (microvasculature) in tendons, using advanced 3D imaging techniques on the equine superficial digital flexor tendon, a model comparable to the human Achilles tendon.
  • The researchers discovered significant reductions in vessel size and density with age, alongside an increase in markers indicating new small blood vessel formation, suggesting complex changes in tendon blood supply during aging.

Background and Motivation

  • Tendon degeneration is a common issue that worsens with age in both humans and horses.
  • Tendons have limited ability to regenerate and heal because they contain few cells and have poor blood supply (vascularisation).
  • Existing treatments often lead to scar tissue formation and a high risk of re-injury, highlighting a need for improved therapeutic strategies.
  • The blood vessels within tendons are crucial for maintaining tendon health (homeostasis), aiding regeneration, and supporting healing after injury.
  • Understanding how aging affects tendon vasculature could reveal new targets for therapies to improve tendon repair, yet this area was previously poorly understood.

Research Objectives

  • To comprehensively characterize the three-dimensional microvasculature of tendons, specifically focusing on age-related changes.
  • To use novel high-resolution imaging techniques — micro-computed tomography (μCT) and confocal microscopy — to visualize and quantify vascular structures in tendons.
  • To focus the study on the equine superficial digital flexor tendon (SDFT), which serves as a functional model for the human Achilles tendon due to similar function and structure.

Methodology

  • Used μCT scans to visualize the overall vascular network within the interfascicular matrix (IFM) of tendons, allowing 3D quantification of vessel volume, diameter, and density.
  • Applied 3D immunolabelling techniques combined with confocal microscopy to detect specific cellular and molecular markers in vessels:
    • MYH11 (Myosin Heavy Chain 11) and desmin, markers associated with vascular smooth muscle cells and vessel stability.
    • Von Willebrand factor (VWF), a marker of endothelial cells involved in blood clotting and vessel formation.
    • Pericyte density, which indicates support cells involved in blood vessel stability and angiogenesis.
  • Compared these vascular characteristics between young and aged tendons to identify age-related changes.

Key Findings

  • μCT imaging showed the tendon has a well-developed vascular network localized in the interfascicular matrix (IFM).
  • Aging caused substantial declines in:
    • Vascular volume by 70%, indicating fewer or smaller vessels overall.
    • Vessel diameter by 30%, meaning vessels become narrower with age.
    • Vessel density by 74%, suggesting less complex vascular networks in aged tendons.
  • Immunolabelling revealed:
    • A 96% reduction in MYH11-positive volume and a 78% reduction in desmin-positive volume, indicating a significant loss of mature, stable blood vessels.
    • A remarkable 220% increase in VWF-positive volume, indicating enhanced presence or activity of endothelial cells, possibly due to new vessel formation.
    • A 249% increase in pericyte density, suggesting that more pericytes are present, which may support newly forming small blood vessels in aged tendons.

Interpretation and Implications

  • The reductions in vascular volume, diameter, and mature vessel markers imply that aging leads to a loss of larger, stable blood vessels within the tendon interfascicular matrix.
  • The increased VWF and pericyte markers suggest stimulated neo-angiogenesis—formation of new small blood vessels—in aged tendons.
  • This dual phenomenon points to disrupted vascular homeostasis with age: while larger vessels deteriorate, the body attempts to compensate by forming new smaller vessels.
  • Understanding these alterations is critical for grasping the mechanisms behind tendon aging and degeneration.
  • The findings may inform the development of therapeutic strategies aimed at restoring healthy blood supply to tendons, potentially improving healing and reducing re-injury risk in older individuals.

Conclusion

  • This study provides the first comprehensive 3D view of tendon microvasculature changes with age.
  • Significant reductions in larger vessels coincide with increased formation of small vessels in aged tendons, indicating altered vascular dynamics.
  • These insights contribute valuable knowledge toward improving tendon health management and therapeutic interventions in aging populations.

Cite This Article

APA
Iwasaki N, Llewellyn J, Brown J, Zamboulis DE, Finding EJT, Wheeler-Jones CPD, Thorpe CT. (2025). Immunolabelling and Micro-Computed Tomography Revealed Age-Related Alterations in 3D Microvasculature of Tendons. Aging Cell, 25(1), e70293. https://doi.org/10.1111/acel.70293

Publication

Aging cell
ISSN: 1474-9726
NlmUniqueID: 101130839
Country: England
Language: English
Volume: 25
Issue: 1
Pages: e70293
PII: e70293

Researcher Affiliations

Iwasaki, Nodoka
  • Comparative Biomedical Sciences, Royal Veterinary College, London, UK.
Llewellyn, Jack
  • Comparative Biomedical Sciences, Royal Veterinary College, London, UK.
Brown, Jeanne
  • Comparative Biomedical Sciences, Royal Veterinary College, London, UK.
Zamboulis, Danae E
  • Department of Clinical Sciences, School of Veterinary Medicine, Aristotle University of Thessaloniki, Thessaloniki, Greece.
Finding, Elizabeth J T
  • Comparative Biomedical Sciences, Royal Veterinary College, London, UK.
Wheeler-Jones, Caroline P D
  • Comparative Biomedical Sciences, Royal Veterinary College, London, UK.
Thorpe, Chavaunne T
  • Comparative Biomedical Sciences, Royal Veterinary College, London, UK.

MeSH Terms

  • Animals
  • X-Ray Microtomography / methods
  • Tendons / blood supply
  • Tendons / diagnostic imaging
  • Tendons / metabolism
  • Aging / physiology
  • Horses
  • Microvessels / diagnostic imaging
  • Microvessels / metabolism
  • Humans
  • Imaging, Three-Dimensional

Grant Funding

  • 804 / Horserace Betting Levy Board

Conflict of Interest Statement

The authors declare no conflicts of interest.

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