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International journal of molecular sciences2025; 26(7); doi: 10.3390/ijms26073353

Extracellular Vesicle-Derived microRNA Crosstalk Between Equine Chondrocytes and Synoviocytes-An In Vitro Approach.

Abstract: This study describes a novel technique to analyze the extracellular vesicle (EV)-derived microRNA (miRNA) crosstalk between equine chondrocytes and synoviocytes. Donor cells (chondrocytes, n = 8; synoviocytes, n = 9) were labelled with 5-ethynyl uridine (5-EU); EVs were isolated from culture media and incubated with recipient cells (chondrocytes [n = 5] were incubated with synoviocyte-derived EVs, and synoviocytes [n = 4] were incubated with chondrocyte-derived EVs). Total RNA was extracted from recipient cells; the 5-EU-labelled RNA was recovered and sequenced. Differential expression analysis, pathway analysis, and miRNA target prediction were performed. Overall, 198 and 213 miRNAs were identified in recipient synoviocytes and chondrocytes, respectively. The top five most abundant miRNAs were similar for synoviocytes and chondrocytes (eca-miR-21, eca-miR-221, eca-miR-222, eca-miR-100, eca-miR-26a), and appeared to be linked to joint homeostasis. There were nine differentially expressed (p < 0.05) miRNAs (eca-miR-27b, eca-miR-23b, eca-miR-31, eca-miR-191a, eca-miR-199a-5p, eca-miR-143, eca-miR-21, eca-miR-181a, and eca-miR-181b) between chondrocytes and synoviocytes, which appeared to be linked to migration of cells, apoptosis, cell viability of connective tissue cell, and inflammation. In conclusion, the reported technique was effective in recovering and characterizing the EV-derived miRNA crosstalk between equine chondrocytes and synoviocytes and allowed for the identification of EV-communicated miRNA patterns potentially related to cell viability, inflammation, and joint homeostasis.
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Publication Date: 2025-04-03 PubMed ID: 40244190PubMed Central: PMC11989968DOI: 10.3390/ijms26073353Google 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.

The research investigates a new method to understand the exchange of microRNA (miRNA) present in extracellular vesicles between horse chondrocytes and synoviocytes, which are all specialized cells relating to joint health. The findings could help shed light on various factors associated with cell vitality, inflammation, and overall joint maintenance.

Methodology

  • The study made use of a newly developed technique to understand the exchange of miRNAs via extracellular vesicles (EVs) between equine chondrocytes and synoviocytes. Chondrocytes are cells found in healthy cartilage, while synoviocytes are cells that line synovial joints, playing essential roles in joint health.
  • The process involved labeling donor cells (chondrocytes and synoviocytes) with 5-ethynyl uridine (5-EU), a compound used for RNA tracking. Extracellular vesicles (EVs) were then isolated from the culture media and incubated with recipient cells.
  • After EV uptake by the recipient cells, total RNA was extracted, and the 5-EU-labeled RNA was subsequently recovered and sequenced.

Analysis and Findings

  • The team performed a differential expression analysis, pathway analysis and miRNA target prediction on the extracted RNA sequences.
  • In the recipient synoviocytes and chondrocytes, 198 and 213 miRNAs were identified, respectively.
  • The five most abundant miRNAs were found to be equivalent across synoviocytes and chondrocytes (eca-miR-21, eca-miR-221, eca-miR-222, eca-miR-100, eca-miR-26a).
  • These miRNAs seemed to be associated with joint homeostasis, or the maintenance of healthy joint function.
  • They found nine miRNAs that were differentially expressed between chondrocytes and synoviocytes. These miRNAs appeared linked to cell apoptosis (programmed cell death), migration of cells, cell viability of connective tissue cell, and inflammation.

Conclusion

  • The technique that the researchers applied was effective in characterizing and recovering the miRNA content of EVs taken up by equine chondrocytes and synoviocytes.
  • The research opened a door to the identification of EV-mediated miRNA patterns potentially related to the vitality of cells, inflammation, and the maintenance of joint health.

Cite This Article

APA
Castanheira CIGD, Anderson JR, Clarke EJ, Hackl M, James V, Clegg PD, Peffers MJ. (2025). Extracellular Vesicle-Derived microRNA Crosstalk Between Equine Chondrocytes and Synoviocytes-An In Vitro Approach. Int J Mol Sci, 26(7). https://doi.org/10.3390/ijms26073353

Publication

International journal of molecular sciences
ISSN: 1422-0067
NlmUniqueID: 101092791
Country: Switzerland
Language: English
Volume: 26
Issue: 7

Researcher Affiliations

Castanheira, Catarina I G D
  • Institute of Life Course and Medical Sciences, University of Liverpool, Liverpool L7 8TX, UK.
Anderson, James R
  • Institute of Life Course and Medical Sciences, University of Liverpool, Liverpool L7 8TX, UK.
  • Institute of Infection, Veterinary and Ecological Sciences, University of Liverpool, Liverpool L3 5RP, UK.
Clarke, Emily J
  • Institute of Life Course and Medical Sciences, University of Liverpool, Liverpool L7 8TX, UK.
Hackl, Matthias
  • TAmiRNA GmbH, 1110 Vienna, Austria.
James, Victoria
  • School of Veterinary Medicine and Science, University of Nottingham, Nottingham LE12 5RD, UK.
Clegg, Peter D
  • Institute of Life Course and Medical Sciences, University of Liverpool, Liverpool L7 8TX, UK.
Peffers, Mandy J
  • Institute of Life Course and Medical Sciences, University of Liverpool, Liverpool L7 8TX, UK.

MeSH Terms

  • Animals
  • MicroRNAs / genetics
  • MicroRNAs / metabolism
  • Horses
  • Chondrocytes / metabolism
  • Chondrocytes / cytology
  • Extracellular Vesicles / metabolism
  • Extracellular Vesicles / genetics
  • Synoviocytes / metabolism
  • Synoviocytes / cytology
  • Cells, Cultured
  • Cell Communication
  • Gene Expression Regulation

Grant Funding

  • Wellcome Trust
  • MR/P020941/1 / Medical Research Council (MRC) and Versus Arthritis as part of the Medical Research Council Versus Arthritis Centre for Integrated Research into Musculoskeletal Ageing
  • 107471/Z/15/Z / Wellcome Trust Clinical Intermediate Fellowship
  • G5018 / The Horse Trust

Conflict of Interest Statement

Matthias Hackl is an employee of TAmiRNA, GmBH. The remaining authors declare no conflicts of interest. The funders had no role in the design of the study; in the collection, analyses, or interpretation of data; in the writing of the manuscript; or in the decision to publish the results.

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