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Frontiers in veterinary science2025; 12; 1671176; doi: 10.3389/fvets.2025.1671176

Metabolomic analysis of synovial fluid from healthy and pathological equine joints and tendon sheaths using high-resolution 1H Nuclear Magnetic Resonance.

Abstract: Joint and tendon sheath diseases are a major cause of lameness and reduced performance in horses. Synovial fluid composition changes in response to pathological processes and metabolomic profiling offers a promising approach to detect these alterations. While equine joint metabolomics has been explored, little is known about the metabolomic profile of tendon sheaths. This study aimed to characterize and compare the synovial fluid metabolomic profiles of healthy and pathological joints and tendon sheaths in horses using high-resolution H Nuclear Magnetic Resonance spectroscopy, and to identify potential biomarkers associated with musculoskeletal pathology. Unassigned: Synovial fluid samples were collected from healthy joints and tendon sheaths of routinely slaughtered animals, and from pathological joints and tendon sheaths from owned athletic horses affected by inflammatory or degenerative conditions. The samples were analyzed using H Nuclear Magnetic Resonance spectroscopy. Synovial fluid samples were collected from healthy joints and tendon sheaths of routinely slaughtered animals, and from pathological joints and tendon sheaths from owned athletic horses affected by inflammatory or degenerative conditions. The samples were analyzed using H Nuclear Magnetic Resonance spectroscopy. Unassigned: The metabolomic analysis of equine synovial fluid identified amino acids, organic acids, glucose isomers, and other metabolites. No significant differences were observed in the metabolic profiles of synovial fluid from healthy joints and tendon sheaths (PCA: RX = 0.761, Q2 = 0.372; OPLS-DA: RX = 0.48; RY = 0.292; Q2 = -0.143). In contrast, a clear separation with distinct clustering was observed between healthy and pathological synovial fluid joints and tendon sheaths (PCA: RX = 0.88, Q2 = 0.684; OPLS-DA: RX = 0.775; RY = 0.6772, Q2 = -0.432). Multivariate statistical analysis revealed distinct clustering of healthy joints samples grouping closely with pathological joints samples (OPLS-DA: RX = 0.662; RX = 0.859, Q2 = 0.786). These findings were supported by univariate analysis (t-test, p < 0.05). Similarly, multivariate statistical analysis showed strong discrimination between healthy and pathological tendon sheaths synovial fluid (OPLS-DA: RX = 0.742; RY = 0.892, Q2 = 0.842), also supported by univariate analysis (t-test,  < 0.05). Unassigned: Metabolomic profiling by H-NMR effectively distinguished healthy from pathological synovial fluid in joints and tendon sheaths, providing a clear metabolic fingerprint of disease-related alterations that may support earlier detection and a better understanding of equine musculoskeletal disorders. The main limitation of this study was the small sample size, particularly for tendon sheath samples. Additional synovial fluid specimens from both healthy and pathological joints and tendon sheaths would be needed to implement metabolomic data. High-resolution H Nuclear Magnetic Resonance spectroscopy proves to be a valuable tool for differentiating healthy from pathological equine synovial fluid. Metabolomic analysis revealed a specific metabolic fingerprint in diseased joints and tendon sheaths, supporting its potential role in the diagnosis and monitoring of orthopedic conditions in horses.
Copyright © 2025 Guadalupi, Girelli, Della Tommasa, Corte, Crovace, Fanizzi, Brehm and Lacitignola.
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Publication Date: 2025-12-16 PubMed ID: 41477162PubMed Central: PMC12747927DOI: 10.3389/fvets.2025.1671176Google Scholar: Lookup
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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 analyzed the chemical composition of synovial fluid from healthy and diseased equine joints and tendon sheaths using high-resolution proton nuclear magnetic resonance (¹H NMR) spectroscopy.
  • The goal was to identify metabolic differences that could serve as biomarkers for musculoskeletal diseases in horses, potentially aiding diagnosis and understanding of joint and tendon sheath disorders.

Background and Importance

  • Joint and tendon sheath diseases are significant causes of lameness and reduced performance in horses, impacting animal welfare and athletic function.
  • Synovial fluid, a lubricating liquid found inside joints and tendon sheaths, changes in its molecular makeup during disease, making it a target for diagnostic biomarker discovery.
  • Metabolomics—the comprehensive analysis of small molecules or metabolites—provides a promising approach to detect these biochemical alterations associated with pathology.
  • While previous research has examined equine joint metabolomics, little was known about the metabolomic profile of tendon sheaths, which are also critical structures affected in musculoskeletal diseases.

Study Design and Methods

  • Synovial fluid samples were collected from:
    • Healthy joints and tendon sheaths from horses slaughtered routinely (serving as controls).
    • Pathological joints and tendon sheaths from athletic horses diagnosed with inflammatory or degenerative conditions.
  • Samples were analyzed using high-resolution ¹H NMR spectroscopy, a technique that detects a range of small molecules based on their unique nuclear magnetic properties.
  • This method allows identification and quantification of multiple metabolites simultaneously, including amino acids, organic acids, and glucose isomers.
  • Multivariate statistical analyses such as Principal Component Analysis (PCA) and Orthogonal Partial Least Squares Discriminant Analysis (OPLS-DA) were used to discern patterns and differences in metabolomic profiles.
  • Univariate statistical tests (t-tests) were also applied to confirm significant differences in individual metabolites.

Key Findings

  • The synovial fluid metabolomic profile included various metabolite classes, notably amino acids, organic acids, and glucose isomers.
  • Comparing healthy joints with healthy tendon sheaths showed no significant metabolic differences, indicating similar baseline metabolite profiles between these tissue types.
  • A clear and statistically significant separation was found between healthy and pathological synovial fluid samples, both in joints and tendon sheaths, based on their metabolomic fingerprints.
  • Statistical analyses demonstrated:
    • Healthy and diseased samples formed distinct clusters, indicating that pathological conditions lead to characteristic metabolic changes.
    • Both PCA and OPLS-DA showed high values of explained variance (R2) and predictive ability (Q2), supporting the robustness of the metabolomic distinctions.
    • Univariate analysis further confirmed significant changes in specific metabolites (p < 0.05), reinforcing the results of multivariate models.

Interpretation and Implications

  • The study demonstrated that high-resolution ¹H NMR metabolomic profiling is effective in distinguishing healthy from pathological synovial fluid in equine joints and tendon sheaths.
  • The unique metabolic fingerprints identified in diseased samples reflect biochemical alterations related to inflammation or degeneration.
  • These metabolic signatures have the potential to:
    • Support earlier and more accurate detection of musculoskeletal diseases in horses.
    • Improve understanding of the underlying biological mechanisms of joint and tendon sheath pathology.
    • Possibly serve as biomarkers for monitoring disease progression or response to therapy.

Limitations and Future Directions

  • The study’s main limitation was the relatively small sample size, especially concerning tendon sheath specimens, which limits the generalizability of the findings.
  • Further research should include larger numbers of synovial fluid samples from both healthy and pathological states to validate and refine the identified metabolic markers.
  • Additional studies could explore longitudinal sampling to track metabolomic changes over time during disease progression or treatment.

Conclusions

  • High-resolution ¹H NMR spectroscopy is a valuable and non-destructive tool for metabolomic analysis of equine synovial fluid.
  • The technique effectively differentiates healthy from diseased joints and tendon sheaths by revealing specific metabolic alterations associated with musculoskeletal pathology.
  • Metabolomic profiling holds promise as a diagnostic aid and a means to enhance understanding of orthopedic conditions in horses, potentially leading to improved clinical outcomes.

Cite This Article

APA
Guadalupi M, Girelli CR, Della Tommasa S, Corte FD, Crovace AM, Fanizzi FP, Brehm W, Lacitignola L. (2025). Metabolomic analysis of synovial fluid from healthy and pathological equine joints and tendon sheaths using high-resolution 1H Nuclear Magnetic Resonance. Front Vet Sci, 12, 1671176. https://doi.org/10.3389/fvets.2025.1671176

Publication

Frontiers in veterinary science
ISSN: 2297-1769
NlmUniqueID: 101666658
Country: Switzerland
Language: English
Volume: 12
Pages: 1671176
PII: 1671176

Researcher Affiliations

Guadalupi, Marta
  • Dipartimento di Medicina di Precisione e Rigenerativa e Area Jonica, Università degli Studi di Bari Aldo Moro, Bari, Italy.
Girelli, Chiara Roberta
  • Department of Biological and Environmental Sciences and Technologies, University of Salento, Lecce, Italy.
Della Tommasa, Simone
  • Department for Horse, University of Leipzig, Leipzig, Germany.
Corte, Federica Della
  • Dipartimento di Medicina di Precisione e Rigenerativa e Area Jonica, Università degli Studi di Bari Aldo Moro, Bari, Italy.
Crovace, Alberto Maria
  • Dipartimento di Medicina Veterinaria, Università di Sassari, Sassari, Italy.
Fanizzi, Francesco Paolo
  • Department of Biological and Environmental Sciences and Technologies, University of Salento, Lecce, Italy.
Brehm, Walter
  • Department for Horse, University of Leipzig, Leipzig, Germany.
Lacitignola, Luca
  • Dipartimento di Medicina di Precisione e Rigenerativa e Area Jonica, Università degli Studi di Bari Aldo Moro, Bari, Italy.

Conflict of Interest Statement

The author(s) declared that this work was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.

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