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BMC medicine2025; 23(1); 439; doi: 10.1186/s12916-025-04231-7

Cryopreserved equine umbilical cord tissue allograft characterization and biocompatibility in vivo in musculoskeletal tissues: a controlled study.

Abstract: The use of micro-particulate allografts is rising, but knowledge about the protein characterization and biocompatibility of umbilical cord-derived allografts (UC) in vivo is limited. Methods: Proteomic analyses using mass spectrometry (MS) determined equine UC protein relative quantification and functions using total spectral counts (TSC). UC cytokines were quantified by enzyme-linked immunosorbent assay (ELISA). Three in vivo studies assessed recipient clinical and tissue biocompatibility in joints and ligaments. Results: Proteomics revealed 2645 annotated TSCs. Proteins of > 89 TSC were considered abundant and were present in all donors. Proteins within the same donor had a 4.7% mean variation. Inflammatory cytokines were low in UC. In vivo, the prospective randomized, masked, controlled study in carpal joints and ligaments of clinically normal horses had median scores of 0 (none) for lameness and pain for 42 days. Synovial fluid showed a transient transudative synovitis after UC injection that was greater than baseline and control and returned to normal after day 5 (P < 0.001). Synovial fluid inflammatory cytokines were low; however, the anti-inflammatory cytokines Il-1ra, Il-10, and Il-1ra/Il-1 ratio were greater after UC injection than at baseline and control (P < 0.001). Blood hematology, chemistries, and serum amyloid A did not reveal systemic effects. The in vivo study of osteoarthritis and desmitis/tendonitis improved in lameness and pain over a 28-day study and had parallel synovial fluid results to the normal horse study, also without adverse events. The in vivo pathologic study evaluated joint and ligament tissues 2 and 5 days after injection and corresponding lymph nodes for evidence of the allograft or inflammation. The synovial membrane, articular cartilage, and lymph nodes were histologically normal, except for mild inflammation in the injection tracts. Conclusions: Well-defined proteins were consistently present in different donors and within batches. Proteins included fibrillar and glycan proteins with a variety of roles and regulatory functions in the connective tissue matrix. The rise in Il-1ra and high Il-1ra/Il-1 ratio after UC injection could block the catabolic effect of Il-1. No adverse events were observed. Within the limits of this study, UC was safe for injection into joints and ligaments in clinically normal horses.
© 2025. The Author(s).
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Publication Date: 2025-07-23 PubMed ID: 40702469PubMed Central: PMC12288288DOI: 10.1186/s12916-025-04231-7Google 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.

The research is about testing the biocompatibility of umbilical cord-derived allografts (UCs) in equine (horse) musculoskeletal tissues. The study reveals that these allografts have a consistent protein composition and can safely be injected into the joints and ligaments without causing adverse effects.

Proteomic Analysis and Cytokine Assessment

  • The research used mass spectrometry, a technique for measuring the mass and concentration of ions, to quantify and understand the function of proteins in the equine umbilical cord (UC).
  • The researchers identified 2645 annotated total spectral counts (TSCs), with proteins having more than 89 TSCs being considered abundant and consistently present across all samples.
  • There was a mean variation of only 4.7% among proteins within a single donor, indicating a high level of consistency.
  • Inflammation-causing cytokines were found to be low in the UC. These cytokines were quantified using enzyme-linked immunosorbent assay (ELISA), a common lab technique to measure antigens in a sample.

In Vivo Biocompatibility Studies

  • Three separate studies were conducted to assess the biocompatibility of UCs when injected into the joints and ligaments of horses.
  • The first, a randomized controlled study, involved injecting the UCs into the carpal (wrist) joints and ligaments of clinically normal horses. These horses displayed no signs of lameness or pain for the duration of the 42-day study.
  • Synovial fluid, a lubricating fluid in joints that reduces friction between cartilage, exhibited a brief increase in inflammation following the injection. However, this returned to normal after five days.
  • The researchers found low levels of inflammatory cytokines in the synovial fluid, but they observed a higher count of specific anti-inflammatory cytokines (Il-1ra and Il-10) after UC injection, suggesting that UC might have an inflammation-dampening effect.

Additional In Vivo Studies

  • The second study focused on horses with osteoarthritis and tendonitis. When treated with UC, these horses demonstrated improved conditions over a 28-day period, without any adverse side effects.
  • The final study investigated the tissues and corresponding lymph nodes for inflammation or other impacts of the UCs two and five days post-injection. The researchers found the synovial membranes, articular cartilage, and lymph nodes to be largely normal, except for mild inflammation.

Conclusions and Implications

  • The study concludes that UC allografts have a consistent protein composition, which includes fibrillar and glycan proteins that play various important roles in the connective tissue matrix.
  • The rise in specific anti-inflammatory cytokines after UC injection suggests that UCs might help to block inflammation, making them potentially useful for treatment in a variety of inflammatory conditions.
  • No adverse effects were observed in any of the in vivo studies, indicating that UCs are safe for injection into horse joints and ligaments.

Cite This Article

APA
Bertone AL, Reinemeyer C, Tsaprailis G, Ragland D, Leise B. (2025). Cryopreserved equine umbilical cord tissue allograft characterization and biocompatibility in vivo in musculoskeletal tissues: a controlled study. BMC Med, 23(1), 439. https://doi.org/10.1186/s12916-025-04231-7

Publication

BMC medicine
ISSN: 1741-7015
NlmUniqueID: 101190723
Country: England
Language: English
Volume: 23
Issue: 1
Pages: 439
PII: 439

Researcher Affiliations

Bertone, Alicia L
  • Department of Veterinary Clinical Sciences (Emeritus), The Ohio State University, 1900 Coffey Rd, Columbus, OH, 43210, USA. Alicia.bertone@gmail.com.
Reinemeyer, Craig
  • East Tennessee Clinical Research, Inc, 80 Copper Ridge Farm Rd, Rockwood, TN, 37854, USA.
Tsaprailis, George
  • The Herbert Wertheim UF Scripps Institute for Biomedical Innovation & Technology; Mass Spectrometry and Proteomics Core Facility, University of Florida, 130 Scripps Way 1B2, Jupiter, FL, 33458, USA.
Ragland, Daniel
  • Veterinary Reference Laboratories, 7540 Louis Pasteur Drive, Suite 200, San Antonio, TX, 78229, USA.
Leise, Britta
  • Department of Veterinary Clinical Sciences, Louisiana State University, Dr Skip Bertman, Baton Rouge, LA, USA.

MeSH Terms

  • Animals
  • Horses
  • Allografts
  • Umbilical Cord / transplantation
  • Cryopreservation
  • Cytokines / analysis
  • Cytokines / metabolism
  • Prospective Studies
  • Female
  • Proteomics
  • Synovial Fluid
  • Male

Conflict of Interest Statement

Declarations. Ethics approval and consent to participate: Animal studies were performed in accordance with and by approval of Institutional Animal Care and Use Committees (ETCR-24-03; LSU-24-089). ARRIVE checklist completed. Consent for publication: NA. Competing interests: ALB was an external consultant for EPL charged to design and manage the research for this study as a third-party contractor due to her publishing and science background in regenerative medicine. She is not an employee and had no equity or other arrangement to benefit from any data outcome.

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