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Home / Equine Research Bank / Equine Vet J / Synovial fluid alpha-2-macroglobulin, gelsolin and lubricin ...
Equine veterinary journal2025; doi: 10.1111/evj.14511

Synovial fluid alpha-2-macroglobulin, gelsolin and lubricin distinguish between osteoarthritic and healthy equine joints.

Abstract: Synovial fluid (SF) is an ideal sentinel fluid for osteoarthritis (OA) diagnosis and prognostication due to its critical homeostatic role, proximity to articular tissues and immune cell composition. Untargeted proteomics enable identification of soluble markers for diagnostic and therapeutic applications while minimising bias. Objective: To use liquid chromatography-tandem mass spectrometry (LC-MS/MS) to define the SF proteome in horses with and without carpal OA. The goal was to identify differentially regulated proteins in mild-moderate carpal joint disease compared with healthy joints. Methods: Cross-sectional study. Methods: Synovial fluid was obtained from horses undergoing arthroscopic treatment for carpal fragmentation and from horses with healthy carpal joints. LC-MS/MS proteomics was performed on a subset of joints (n = 8 OA, n = 8 healthy). Total protein (TP), gelsolin, lubricin and prostaglandin E (PGE) concentrations were quantified via biochemical or immunoassays (n = 58 OA, n = 25 healthy), and synovial membrane histology was graded (n = 16 OA, n = 6 healthy). Univariate and multivariate modelling were used to compare the predictive capacities of soluble factors between healthy and OA joints. Results: Of 119 proteins identified, 14 were increased and 10 decreased in OA. Two of the most upregulated proteins included pregnancy zone protein and alpha-2-macroglobulin. In OA, gelsolin/TP was decreased (OA median: 0.48 μg/mL/mg [95% confidence interval 0.27-0.73]; healthy: 0.89 μg/mL/mg [0.63-1.17]) and lubricin/TP increased (OA: 27.98 μg/mL/mg, [21.71-45.79]; healthy: 6.77 μg/mL/mg [4.36-9.94]) when measured by immunoassay. Multivariate modelling including gelsolin/TP, lubricin/TP and PGE was superior to univariate models for differentiating between OA and healthy joints. Conclusions: Synovial membrane was available for a limited number of joints and most healthy samples were collected following euthanasia. Conclusions: The upregulation of pregnancy zone protein and alpha-2-macroglobulin in OA joints motivates investigation into their function in OA and development of reagents for quantification. Several proteins with differential abundance in OA SF, including gelsolin and lubricin, were measured and may have diagnostic utility. Unassigned: Synovialflüssigkeit (SF) ist aufgrund ihrer kritischen homöostatischen Rolle, ihrer Nähe zu den Gelenkgeweben und ihrer Leukozytenzusammensetzung eine ideale Sentinel‐Flüssigkeit für die Diagnose und Prognose von Osteoarthritis (OA). Ungezielte Proteomik ermöglicht die Identifizierung löslicher Marker für diagnostische und therapeutische Anwendungen bei gleichzeitiger Minimierung von Verzerrungen. Unassigned: Einsatz der Flüssigchromatographie‐Tandem‐Massenspektrometrie (LC‐MS/MS) zur Bestimmung des SF‐Proteoms bei Pferden mit und ohne Karpalgelenksosteoarthritis. Ziel war es, unterschiedlich regulierte Proteine bei Leichter bis mittelschwerer Karpalgelekserkrankung im Vergleich zu gesunden Gelenken zu identifizieren. Methods: Bi‐direktionale Fall‐Kontroll‐Studie. Methods: SF wurde von Pferden gewonnen, die sich einer arthroskopischen Behandlung wegen Karpalfragmentierung unterzogen, sowie von Pferden mit gesunden Karpalgelenken. LC‐MS/MS‐Proteomik wurde an einer Untergruppe von Gelenken durchgeführt (n=8 OA, n=8 gesunde). Die Konzentrationen von Gesamtprotein (TP), Gelsolin, Lubricin und Prostaglandin E2 (PGE2) wurden mit biochemischen oder Immunoassays quantifiziert (n=58 OA, n=25 Gesunde), und die Histologie der Synovialmembran wurde bewertet (n=16 OA, n=6 Gesunde). Mittels univariater und multivariater Modellierung wurde die Vorhersagekraft löslicher Faktoren zwischen gesunden und OA‐Gelenken verglichen. Unassigned: Von 119 identifizierten Proteinen waren 14 bei OA erhöht und 10 vermindert. Zwei der am stärksten erhöhten Proteine waren das pregnancy zone Protein und Alpha‐2‐Makroglobulin. Bei OA war Gelsolin/TP vermindert (OA‐Median: 0,48μg/mL/mg (CI 95% 0,27‐0,73); gesund: 0,89μg/mL/mg (0,63‐1,17)) und Lubricin/TP erhöht (OA: 27,98μg/mL/mg, (21,71‐45,79); gesund: 6,77μg/mL/mg (4,36‐9,94)) wenn gemessen mittels Immunoassay. Die multivariate Modellierung unter Einbeziehung von Gelsolin/TP, Lubricin/TP und PGE2 war den univariaten Modellen zur Unterscheidung zwischen OA und gesunden Gelenken überlegen. HAUPTEINSCHRÄNKUNGEN: Synovialmembran war nur für eine begrenzte Anzahl von Gelenken verfügbar, und die meisten gesunden Proben wurden nach der Euthanasie entnommen. Unassigned: Die Hochregulierung von pregnancy zone Protein und Alpha‐2‐Makroglobulin in OA‐Gelenken motiviert zur Untersuchung ihrer Funktion bei OA und zur Entwicklung von Reagenzien für die Quantifizierung. Mehrere Proteine mit unterschiedlicher Häufigkeit in OA‐SF, einschließlich Gelsolin und Lubricin, wurden gemessen und könnten einen diagnostischen Nutzen haben.
© 2025 The Author(s). Equine Veterinary Journal published by John Wiley & Sons Ltd on behalf of EVJ Ltd.
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Publication Date: 2025-05-08 PubMed ID: 40342270DOI: 10.1111/evj.14511Google Scholar: Lookup
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Summary

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The research investigates the protein content of synovial fluid in horses with and without osteoarthritis (OA) for potential diagnostic markers of the disease. Liquid chromatography-tandem mass spectrometry (LC-MS/MS) was used to analyze the protein content of these samples, which indicated changes in specific proteins in the horses with OA.

Objective of the Study

The primary objective of this research was to identify differences in the protein content of synovial fluid, the joint lubricating fluid, in horses with OA compared to healthy horses. The goal was to discover potential biomarkers for OA.

Methods

  • The researchers collected synovial fluid from horses undergoing treatment for carpal fragmentation (a form of OA in horse’s carpal joints) and from horses with healthy joints.
  • A subset from these samples (8 with OA, 8 healthy) was analyzed using Liquid Chromatography and Tandem Mass Spectrometry (LC-MS/MS). The technique enabled a detailed analysis of the proteins present in the fluid.
  • Total protein concentration and the concentrations of specific proteins (gelsolin, lubricin, and Prostaglandin E) were measured using biochemical and immunoassay methods.
  • The researchers also graded the synovial membrane histology for a number of joints (for both OA and healthy samples).
  • Statistical methods were used to compare the protein content of the OA and healthy joints and measure the predictive capacities of the proteins discovered.

Results

  • The LC-MS/MS analysis identified 119 proteins. Among them, 14 proteins were found in higher concentrations, and 10 were found in lower concentrations in the synovial fluid from horses with OA.
  • The most upregulated proteins were a pregnancy zone protein and alpha-2-macroglobulin.
  • In cases of OA, the concentration of gelsolin per total protein was decreased, while the concentration of lubricin per total protein was increased.
  • When multivariate modeling including gelsolin, lubricin, and prostaglandin E was used, it was found to be better at differentiating between OA and healthy joints compared to univariate modelling.

Conclusions

  • The research thus indicates that protein content in synovial fluid, specifically the levels of the pregnancy zone protein, alpha-2-macroglobulin, gelsolin, and lubricin, could potentially be used as diagnostic tools for osteoarthritis in horses.
  • The changes in these specific proteins imply they may play a role in the development of OA, and further investigation into their function in the disease may provide new insights into potential therapies.

Cite This Article

APA
Secor EJ, Womack SJ, Ysebaert MP, Colville MJ, Reesink HL. (2025). Synovial fluid alpha-2-macroglobulin, gelsolin and lubricin distinguish between osteoarthritic and healthy equine joints. Equine Vet J. https://doi.org/10.1111/evj.14511

Publication

Equine veterinary journal
ISSN: 2042-3306
NlmUniqueID: 0173320
Country: United States
Language: English

Researcher Affiliations

Secor, Erica J
  • Department of Clinical Sciences, Cornell University College of Veterinary Medicine, Cornell University, Ithaca, New York, USA.
Womack, Sydney J
  • Department of Clinical Sciences, Cornell University College of Veterinary Medicine, Cornell University, Ithaca, New York, USA.
Ysebaert, Machiel P
  • Department of Clinical Sciences, Cornell University College of Veterinary Medicine, Cornell University, Ithaca, New York, USA.
  • Department of Clinical Sciences, Colorado State University College of Veterinary Medicine and Biomedical Sciences, Colorado State University, Fort Collins, Colorado, USA.
Colville, Marshall J
  • Department of Clinical Sciences, Cornell University College of Veterinary Medicine, Cornell University, Ithaca, New York, USA.
  • Robert Frederick Smith School of Chemical and Biomolecular Engineering, Cornell University, Ithaca, New York, USA.
Reesink, Heidi L
  • Department of Clinical Sciences, Cornell University College of Veterinary Medicine, Cornell University, Ithaca, New York, USA.
  • Department of Surgical & Radiological Sciences, School of Veterinary Medicine, University of California-Davis, Davis, California, USA.

Grant Funding

  • Hong Kong Jockey Club Equine Welfare Research Foundation
  • TR002384 / Clinical and Translational Science Center, Weill Cornell Medical College
  • American College of Veterinary Surgeons Foundation

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