Stimulus-dependent release of tissue-regenerating factors by equine platelets.
Abstract: Platelet-rich plasma (PRP) is increasingly used for treatment of orthopaedic injuries. However, the effects of different stimuli on the release pattern of regenerative and proinflammatory factors from equine platelets are largely unknown and an optimal treatment protocol remains to be established. Objective: The aim of this study was to identify a stimulus that enhanced release of histopromotive factors (platelet-derived growth factor BB [PDGF] and transforming growth factor 1β[TGF]) without causing concurrent release of a proinflammatory mediator (CCL5). Methods: Washed platelets were prepared from 6 healthy ponies and release of growth factors and CCL5 measured using commercially available ELISAs for human proteins following incubation with or without thrombin, chitosan or equine recombinant tumour necrosis factor (erTNF) over 24 h and subsequently over 96 h. Additionally, noncoagulated samples were analysed. Results: Regardless of whether a stimulus was present or what stimulus was used, PDGF and TGF release was maximal by 0.5-1 h when clot formation took place and very little release was observed after 24 h. Growth factor release was minimal in noncoagulated samples. In contrast, CCL5 release was not associated with coagulation and appeared to persist for much longer. High concentrations of erTNF caused significantly greater release of CCL5 at 6 h than any other stimulus tested. Conclusions: Growth factor release from equine platelets is dependent on coagulation but independent of the initiating stimulus, and is accompanied by more sustained release of proinflammatory mediators. Conclusions: Supernatants collected from coagulated platelets could be an alternative treatment to PRP.
© 2011 EVJ Ltd.
Publication Date: 2011-09-09 PubMed ID: 21902713DOI: 10.1111/j.2042-3306.2011.00431.xGoogle Scholar: Lookup
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- Journal Article
- Research Support
- Non-U.S. Gov't
Summary
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This research investigates how different stimuli impact the release of regenerative and proinflammatory factors from equine platelets. This could help refine treatment protocols for orthopaedic injuries with platelet-rich plasma (PRP). The tests were initiated with or without thrombin, chitosan, or equine recombinant tumor necrosis factor (erTNF) and observed over 24 and 96 hour periods.
Methods
- Equine platelets were prepared from 6 healthy ponies.
- The release of growth factors (PDGF and TGF) and a proinflammatory mediator (CCL5) was measured.
- Commercial ELISAs for human proteins were used to measure the release following incubation with or without thrombin, chitosan, or erTNF.
- The measurement was carried out over 24h and subsequently over 96h.
- Noncoagulated samples were also analyzed.
Results
- The release of growth factors (PDGF and TGF) was observed to be maximal between 0.5 – 1h when clot formation occurred, irrespective of whether a stimulus was present or not. It was also noted that very little release occurred after 24h.
- It was observed that these growth factors were minimally released in noncoagulated samples.
- CCL5 release, on the other hand, did not associate with coagulation and was observed to persist much longer.
- High concentrations of erTNF resulted in considerably greater release of CCL5 at 6h compared to other stimuli tested.
Conclusions
- It was concluded that growth factor release from equine platelets is dependent on coagulation but independent of the initiating stimulus. This release was noted to be accompanied by a more sustained release of proinflammatory mediators.
- Another key conclusion from the study was that supernatants collected from coagulated platelets could potentially serve as an alternative treatment to PRP.
Cite This Article
APA
Dunkel B, Bolt DM, Smith RK, Cunningham FM.
(2011).
Stimulus-dependent release of tissue-regenerating factors by equine platelets.
Equine Vet J, 44(3), 346-354.
https://doi.org/10.1111/j.2042-3306.2011.00431.x Publication
Researcher Affiliations
- Departments of Veterinary Clinical Sciences and Veterinary Basic Sciences, The Royal Veterinary College, UK.
MeSH Terms
- Animals
- Blood Platelets / drug effects
- Blood Platelets / metabolism
- Cells, Cultured
- Chemokine CCL5 / genetics
- Chemokine CCL5 / metabolism
- Chitosan / pharmacology
- Gene Expression Regulation / drug effects
- Horses / blood
- Horses / metabolism
- Intercellular Signaling Peptides and Proteins / genetics
- Intercellular Signaling Peptides and Proteins / metabolism
- Platelet Activation / drug effects
- Platelet Activation / physiology
- Platelet-Derived Growth Factor / genetics
- Platelet-Derived Growth Factor / metabolism
- Thrombin / pharmacology
- Transforming Growth Factors / genetics
- Transforming Growth Factors / metabolism
- Tumor Necrosis Factor-alpha / pharmacology
Citations
This article has been cited 4 times.- McClain AK, McCarrel TM. The effect of four different freezing conditions and time in frozen storage on the concentration of commonly measured growth factors and enzymes in equine platelet-rich plasma over six months. BMC Vet Res 2019 Aug 14;15(1):292.
- Seabaugh KA, Thoresen M, Giguère S. Extracorporeal Shockwave Therapy Increases Growth Factor Release from Equine Platelet-Rich Plasma In Vitro. Front Vet Sci 2017;4:205.
- Jalowiec JM, D'Este M, Bara JJ, Denom J, Menzel U, Alini M, Verrier S, Herrmann M. An In Vitro Investigation of Platelet-Rich Plasma-Gel as a Cell and Growth Factor Delivery Vehicle for Tissue Engineering. Tissue Eng Part C Methods 2016 Jan;22(1):49-58.
- Di Ianni F, Merli E, Burtini F, Conti V, Pelizzone I, Di Lecce R, Parmigiani E, Squassino GP, Del Bue M, Lucarelli E, Ramoni R, Grolli S. Preparation and application of an innovative thrombocyte/leukocyte-enriched plasma to promote tissue repair in chelonians. PLoS One 2015;10(4):e0122595.
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