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Equine veterinary journal2024; doi: 10.1111/evj.14090

Equine bone marrow MSC-derived extracellular vesicles mitigate the inflammatory effects of interleukin-1β on navicular tissues in vitro.

Abstract: Safe, efficacious therapy for treating degenerate deep digital flexor tendon (DDFT) and navicular bone fibrocartilage (NBF) in navicular horses is critically necessary. While archetypal orthobiologic therapies for navicular disease are used empirically, their safety and efficacy are unknown. Mesenchymal stem cell-derived extracellular vesicles (EV) may overcome several limitations of current orthobiologic therapies. Objective: To (1) characterise cytokine and growth factor profiles of equine bone marrow mesenchymal stem cell (BM-MSC)-derived extracellular vesicles (BM-EV) and (2) evaluate the in vitro anti-inflammatory and extracellular matrix (ECM) protective potentials of BM-EV on DDFT and NBF explant co-cultures in an IL-1β inflammatory environment. Methods: In vitro experimental study. Methods: Cytokines (IL-1β, IL-6, IL-10, IL-1ra and TNF-α) and growth factors (TGFβ1, VEGF, IGF1 and PDGF) in equine BM-EV isolated via ultracentrifugation and precipitation methods were profiled. Forelimb DDFT and NBF explant co-cultures from seven horses were exposed to media alone, or media containing 2 × 10 ± 0.1 × 10 particles/mL or 10 μg/mL BM-EV (BM-EV), 10 ng/mL interleukin-1β (IL-1β), or IL-1β + BM-EV for 48 h. Co-culture media IL-6, TNF-α, MMP-3, MMP-13 concentrations and explant sulphated glycosaminoglycan (sGAG) content were quantified. Results: IL-6, IGF1 and VEGF concentrations were 102.1 (37.61-256.2) and 182.3 (163.1-226.3), 72.3 (8-175.6) and 2.4 (0.1-2.6), 108.3 (38.3-709.1) and 211.4 (189.1-318.2) pg/mL per 2 × 10 ± 0.1 × 10 particles/mL or 10 μg/mL 10 μg of BM-EV isolated via ultracentrifugation and precipitation methods, respectively. Co-culture media MMP-3 in BM-EV- (p = 0.03) and BM-EV + IL-1β-treated (p = 0.01) groups were significantly lower than the respective media and IL-1β groups. DDFT explant sGAG content of BM-EV (p = 0.003) and BM-EV + IL-1β groups were significantly higher compared with IL-1β group. Conclusions: Specimen numbers are limited, in vitro model may not replicate clinical case conditions, lack of non-MSC-derived EV control group. Conclusions: Equine BM-EV contains IL-6 and growth factors, IGF1 and VEGF. The anti-inflammatory and ECM protective potentials of BM-EV were evident as increased IL-6 and decreased MMP-3 concentrations in the DDFT-NBF explant co-culture media. These results support further evaluation of BM-EV as an acellular and 'off-the-shelf' intra-bursal/intrasynovial therapy for navicular pathologies.
© 2024 The Authors. Equine Veterinary Journal published by John Wiley & Sons Ltd on behalf of EVJ Ltd.
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Publication Date: 2024-04-08 PubMed ID: 38587145DOI: 10.1111/evj.14090Google 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.

This study investigated the anti-inflammatory and protective effects of extracellular vesicles (EVs) from equine bone marrow mesenchymal stem cells (BM-MSCs) on horse tendon and navicular bone tissue in the presence of a common inflammatory mediator.

Objectives and Methods

  • The research had two primary goals:
    1. To identify the different types of cytokines (a type of protein involved in cell signaling) and growth factors present in BM-EV.
    2. To test the potential of BM-EV to counter inflammation and protect the extracellular matrix (ECM, a network of molecules that provides physical and biochemical support to cells) in deep digital flexor tendon and navicular bone fibrocartilage tissues exposed to the inflammatory cytokine interleukin-1β (IL-1β).
  • This was an experimental in vitro study, meaning it was performed in a controlled laboratory environment (as opposed to in a live organism).
  • The researchers isolated BM-EV from equine BM-MSCs and analyzed their content of several cytokines and growth factors.
  • Explant co-cultures (isolated cells or tissues grown together) of deep digital flexor tendon and navicular bone fibrocartilage from seven horses were treated with either BM-EVs, IL-1β, or both, and the subsequent inflammatory and structural responses were examined.

Results

  • The BM-EV samples were found to contain il-6, IGF1, and VEGF, among other factors.
  • The researchers noted changes in the levels of IL-6 and MMP-3, markers of inflammation and tissue degradation, respectively, in the co-cultures treated with BM-EVs. While IL-6 levels were found to increase, MMP-3 levels were found to decrease. These alterations would suggest an anti-inflammatory response as well as a potential protection against tissue damage.
  • The content of sulphated glycosaminoglycan (a type of molecule involved in building the ECM) in the treated explants was reported to be higher than in the IL-1β-only control group, suggesting that the BM-EVs could have a protective effect on the structure of the tissues.

Limits and Conclusions

  • The study had several limitations, including limited sample size, the fact that the in vitro model may not accurately reflect the complex conditions inside a living organism, and the lack of a control group treated with non-MSC-derived EV.
  • Despite these limitations, the authors concluded that equine BM-EVs seem to contain anti-inflammatory and growth-promoting factors and that their application mitigated inflammation and potentially protected tissue integrity in the tested explant co-cultures.
  • These results suggest that BM-EVs could be a potentially useful therapy for treating inflammatory conditions in equine tendons and bones, but the authors stressed the need for further study.

Cite This Article

APA
Quam VG, Belacic ZA, Long S, Rice HC, Dhar MS, Durgam S. (2024). Equine bone marrow MSC-derived extracellular vesicles mitigate the inflammatory effects of interleukin-1β on navicular tissues in vitro. Equine Vet J. https://doi.org/10.1111/evj.14090

Publication

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

Researcher Affiliations

Quam, Vivian G
  • Department of Veterinary Clinical Sciences, College of Veterinary Medicine, The Ohio State University, Columbus, Ohio, USA.
  • Ballarat Veterinary Practice Equine Clinic, Miners Rest, Victoria, Australia.
Belacic, Zarah A
  • Department of Veterinary Clinical Sciences, College of Veterinary Medicine, The Ohio State University, Columbus, Ohio, USA.
Long, Sidney
  • Department of Veterinary Clinical Sciences, College of Veterinary Medicine, The Ohio State University, Columbus, Ohio, USA.
Rice, Hilary C
  • Department of Veterinary Clinical Sciences, College of Veterinary Medicine, The Ohio State University, Columbus, Ohio, USA.
Dhar, Madhu S
  • Department of Large Animal Clinical Sciences, College of Veterinary Medicine, University of Tennessee, Knoxville, Tennessee, USA.
Durgam, Sushmitha
  • Department of Veterinary Clinical Sciences, College of Veterinary Medicine, The Ohio State University, Columbus, Ohio, USA.

Grant Funding

  • KL2TR002734 / NCATS NIH HHS

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Citations

This article has been cited 3 times.
  1. Najeb M, Samy A, Rizk A, Mosbah E, Karrouf G. Regenerative biologics modulating inflammation and promoting tenogenesis in equine superficial digital flexor tendonitis: from molecular pathways to clinical translation. Ir Vet J 2025 Sep 17;78(1):21.
    doi: 10.1186/s13620-025-00309-zpubmed: 40963139google scholar: lookup
  2. Aeri A, Gorla M, Sharma GT. Veterinary Regenerative Medicine: The Evolving Role of Stem Cell-Based Therapies. Stem Cell Rev Rep 2025 Nov;21(8):2484-2510.
    doi: 10.1007/s12015-025-10963-zpubmed: 40900287google scholar: lookup
  3. Milczek-Haduch D, Żmigrodzka M, Witkowska-Piłaszewicz O. Extracellular Vesicles in Sport Horses: Potential Biomarkers and Modulators of Exercise Adaptation and Therapeutics. Int J Mol Sci 2025 May 3;26(9).
    doi: 10.3390/ijms26094359pubmed: 40362597google scholar: lookup
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