Abstract
BackgroundThere is a lack information on the effects of the most commonly used anticoagulants for equine platelet rich plasmas (PRPs) elaboration on cell counts and growth factor release from platelet rich gels (PRGs). The aims of this study were 1) to compare the effects of the anticoagulants sodium citrate (SC), acid citrate dextrose solution A (ACD-A) and ACD-B on platelet (PLT), leukocyte (WBC) and on some parameters associated to platelet activation including mean platelet volume (MPV) and platelet distribution width (PDW) between whole blood, pure PRP (P-PRP) and platelet-poor plasma (PPP); 2) to compare transforming growth factor beta 1 (TGF-β1) and platelet-derived growth factor isoform BB (PDGF-BB) concentrations in supernatants from pure PRG (P-PRG), platelet-poor gel (PPG), P-PRP lysate (positive control) and plasma (negative control); 3) to establish the possible correlations between all the studied cellular and molecular parameters.ResultsIn all cases the three anticoagulants produced P-PRPs with significantly higher PLT counts compared with whole blood and PPP. The concentrations of WBCs were similar between P-PRP and whole blood, but significantly lower in PPP. The type of anticoagulant did not significantly affect the cell counts for each blood component. The anticoagulants also did not affect the MPV and PDW parameters. Independently of the anticoagulant used, all blood components presented significantly different concentrations of PDGF-BB and TGF-β1. The highest growth factor (GF) concentrations were observed from P-PRP lysates, followed by PRG supernatants, PPP lysates, PPG supernatants and plasma. Significant correlations were observed between PLT and WBC counts (ρ = 0.80), PLT count and TGF-β1 concentration (ρ = 0.85), PLT count and PDGF-BB concentration (ρ = 0.80) and PDGF-BB and TGF-β1 concentrations (ρ = 0.75). The type of anticoagulant was not correlated with any of the variables evaluated.ConclusionsThe anticoagulants did not significantly influence cell counts or GF concentrations in equine PRP. However, ACD-B was apparently the worst anticoagulant evaluated. It is necessary to perform additional research to determine the effect of anticoagulants on the kinetics of GF elution from P-PRG.
Highlights
There is a lack information on the effects of the most commonly used anticoagulants for equine platelet rich plasmas (PRPs) elaboration on cell counts and growth factor release from platelet rich gels (PRGs)
The highest growth factor (GF) concentrations were observed from pure-platelet rich plasma (P-platelet-rich plasma (PRP)) lysates, followed by PRG supernatants, Platelet poor plasma (PPP) lysates, Platelet poor plasma gel (PPG) supernatants and plasma (Table 2)
Correlations Significant correlations were observed between PLT and WBC counts (ρ = 0.80, P
Summary
There is a lack information on the effects of the most commonly used anticoagulants for equine platelet rich plasmas (PRPs) elaboration on cell counts and growth factor release from platelet rich gels (PRGs). It was recognized that among the regenerative therapies, platelet-rich plasma (PRP) is an autologous platelet concentrate suspended in plasma that, administrated in the wound site, releases growth factors and promotes the wound healing cascade [2,5]. Platelets contain a pool of growth factors, including transforming growth factor-b (TGF-β), platelet derived growth factor (PDGF) and vascular endothelial growth factor (VEGF), mainly contained in platelet alpha granules [6] that are released after platelet degranulation in the damage site and enhance tissue regeneration by stimulating cell proliferation, increasing extracellular matrix synthesis, promoting vascular ingrowth and reducing catabolic matrix-degrading cytokines such as interleukins and matrix metalloproteinases [5,7]. In vitro evidence suggests that leukoreduced PRP could be more suitable for the treatment of tendon and soft tissue injuries in horses, as this substance induces tendon anabolism and decreases the expression of catabolic cytokines when compared with L-PRP [9]
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