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Autologous conditioned serum: the comparative cytokine profiles of two commercial methods (IRAP and IRAP II) using equine blood.
Abstract: Osteoarthritis (OA) is one of the most prevalent and debilitating conditions affecting the horse. Autologous conditioned serum (ACS), commercially available as IRAP and IRAP II, is a recently developed treatment for OA in which plasma is prepared from venous blood by incubation with glass beads for 24 h. This product has been shown to increase anti-inflammatory cytokines and growth factors in human blood. However, data for equine ACS preparations are lacking. Objective: To characterise the protein profiles produced by commercially available ACS systems in equine blood. Methods: Blood was drawn from 5 horses into 6 groups: red top vacutainer (control), IRAP and IRAP II, with and without heparin. Samples were collected 1 or 24 h post draw and analysed for IL-1ra, IL-10, IGF-1, TGF-β, TNF-α and IL-1β using ELISAs. Results: Twenty-four hour IRAP and IRAP II samples contained significantly higher levels of all cytokines relative to 1 h serum controls. At 24 h, IRAP II contained significantly higher levels of IL-1ra and IRAP contained significantly higher levels of TNF-α, compared to 24 h controls. In addition, TGF-β, IL-10 and IL-1β in IRAP and IRAP II sera were similar to 24 h serum controls. The addition of heparin significantly reduced levels of IGF-1, TNF-α and TGF-β, and significantly elevated levels of IL-1ra. Conclusions: The cytokine profile that IRAP II produced is modestly better than IRAP. Incubation of whole blood in glass tubes stimulated cytokine synthesis, although not as efficiently as IRAP II. Conclusions: Although high levels of IL-1ra were found in ACS, elevation of other factors suggests these cytokines play a previously understated role in clinical improvements. Because ACS has been shown to alleviate clinical symptoms of OA, the present study suggests that factors other than IL-1ra alone might be involved in its clinical efficacy. Species-dependent elevations of cytokines warrant further investigation and optimisation of the systems appears to be necessary based on the differences between human and equine blood.
© 2011 EVJ Ltd.
Publication Date: 2011-03-11 PubMed ID: 21496084DOI: 10.1111/j.2042-3306.2010.00321.xGoogle Scholar: Lookup The Equine Research Bank provides access to a large database of publicly available scientific literature. Inclusion in the Research Bank does not imply endorsement of study methods or findings by Mad Barn.
- Journal Article
- Research Support
- Non-U.S. Gov't
Summary
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The research investigates the effect of two commercially available treatments – IRAP and IRAP II – on horse blood to compare their efficiency for treating Osteoarthritis (OA). The study found that both treatments boosted anti-inflammatory cytokines and growth factors, with IRAP II producing marginally better results than IRAP.
Methodology
- The study used blood samples drawn from five horses, which were divided into six groups: red top vacutainer (control), IRAP and IRAP II, applied both with and without the addition of heparin.
- Samples were collected at 1 hour or 24 hours after the draw and were analysed for different cytokines (proteins impacting cell behaviour) using Enzyme-Linked Immunosorbent Assays (ELISAs). The measured cytokines included IL-1ra, IL-10, IGF-1, TGF-β, TNF-α and IL-1β.
Results
- Data showed that 24-hour IRAP and IRAP II samples had significantly higher levels of all cytokines compared to the 1-hour controls.
- At the 24-hour mark, IRAP II samples contained notably higher levels of IL-1ra, a protein that regulates inflammation, while IRAP samples had higher levels of TNF-α, a cytokine involved in inflammatory reactions.
- Meanwhile, the levels of TGF-β, IL-10 and IL-1β were found to be similar in both IRAP and IRAP II comparisons to control.
- The addition of heparin (a medication that prevents blood clots) decreased levels of IGF-1, TNF-α and TGF-β, and increased levels of IL-1ra.
Conclusions
- The study indicates that even though IRAP II produced a slightly better cytokine profile than IRAP, both treatments increase cytokine synthesis in equine blood.
- Additionally, the study suggests that not only high IL-1ra levels, but also other related factors, might play a role in clinical improvements relating to OA therapy.
- Due to observed species-dependent variations in cytokine levels, further investigation and optimization of these anti-OA systems is recommended, especially considering differences between human and equine blood.
Cite This Article
APA
Hraha TH, Doremus KM, McIlwraith CW, Frisbie DD.
(2011).
Autologous conditioned serum: the comparative cytokine profiles of two commercial methods (IRAP and IRAP II) using equine blood.
Equine Vet J, 43(5), 516-521.
https://doi.org/10.1111/j.2042-3306.2010.00321.x Publication
Researcher Affiliations
- Gail Holmes Equine Orthopaedic Research Center, Colorado State University, Colorado, USA.
MeSH Terms
- Animals
- Blood Transfusion, Autologous / veterinary
- Cytokines / genetics
- Cytokines / metabolism
- Enzyme-Linked Immunosorbent Assay / veterinary
- Gene Expression Regulation
- Horses / blood
- Interleukin-1 Receptor Accessory Protein / metabolism
- RNA, Messenger / genetics
- RNA, Messenger / metabolism
- Real-Time Polymerase Chain Reaction / veterinary
- Serum / chemistry
Citations
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