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. 2016 Mar 7;2(1):e000071.
doi: 10.1136/bmjsem-2015-000071. eCollection 2016.

Normal Platelet Function in Platelet Concentrates Requires Non-Platelet Cells: A Comparative in Vitro Evaluation of Leucocyte-Rich (Type 1a) and Leucocyte-Poor (Type 3b) Platelet Concentrates

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Free PMC article

Normal Platelet Function in Platelet Concentrates Requires Non-Platelet Cells: A Comparative in Vitro Evaluation of Leucocyte-Rich (Type 1a) and Leucocyte-Poor (Type 3b) Platelet Concentrates

William R Parrish et al. BMJ Open Sport Exerc Med. .
Free PMC article

Abstract

Background: Therapeutic success of platelet-rich plasma (PRP) may vary based on the composition and preparation method. The objective of this study was to evaluate the cellular components of platelet concentrates produced by a leucocyte-rich (LR-PRP) and a leucocyte-poor PRP systems (LP-PRP).

Methods: Parameters evaluated included platelet recovery, platelet concentration, red blood cell (RBC) and white blood cell (WBC) composition, platelet growth factor release and stimulation of human tendon cell proliferation in vitro.

Results: Platelet recoveries were 52% for LP-PRP and 89% for LR-PRP. LR-PRP demonstrated greater reproducibility with a 4.2% coefficient of variation (CV) compared with 19.4% for LP-PRP (p<0.001). LR-PRP demonstrated a greater increase in platelet concentration (7.9-fold) than LP-PRP (2.2-fold; p<0.001). LP-PRP showed 5.0-fold reductions in WBCs, while LR-PRP showed a 4.0-fold increase (p<0.001). LP-PRP reduced RBCs to a haematocrit of 0.25, while LR-PRP reduced haematocrit to 11.8. LP-PRP did not coagulate robustly on reactivation with CaCl2, and released significantly lower levels of epidermal growth factor (EGF) and transforming growth factor β1 (TGF-β1) than whole blood (p<0.03). LP-PRP also did not stimulate tendon cell proliferation greater than whole blood. In contrast, LR-PRP showed increases in each growth factor on activation with CaCl2 (p<0.01) and stimulated greater proliferation (p<0.05) compared with whole blood. Forced activation of LP-PRP with exogenous thrombin rescued the coagulation deficiency and induced greater growth factor release than comparable whole blood (p<0.03).

Conclusions: These data suggest that non-platelet cellular components in platelet concentrates are important for proper platelet function, including thrombin generation, growth factor release and clot retraction.

Keywords: Haematology; Plasma; Platelet-Rich Plasma.

Figures

Figure 1
Figure 1
Fold change in platelet concentration. Bars represent the mean ± SD of the fold increase in platelet content (n=12 donors). Fold increase=platelet count in platelet concentrate/whole blood platelet count.
Figure 2
Figure 2
Platelet recovery. Bars represent the mean ± SD of the percentage of total platelets recovered in platelet concentrates (n=12 donors). Per cent platelet recovery=(total platelet count of platelet concentrate (concentration × total volume)/total platelet count of whole blood (concentration × total volume))×100.
Figure 3
Figure 3
Stimulation of tendon cell proliferation. Data represent the mean ± SD of preparations from five individual donors. All preparations from each donor were assayed in parallel using reactivated platelet-poor plasma as the baseline control for each serum concentration tested.
Figure 4
Figure 4
Serum growth factor release. Platelet-poor plasma, whole blood and platelet concentrates from the LP-PRP and LR-PRP systems were reactivated with a final concentration of 25 mM CaCl2 for 80 min at room temperature. Growth factor levels were measured in the resulting serum by ELISA. Bars represent the mean ± SD from five independent donors. All preparations from each donor were assayed in parallel. LP-PRP, leucocyte-poor platelet-rich plasma; LR-PRP, leucocyte-rich platelet-rich plasma.
Figure 5
Figure 5
Growth factor release from LP-PRP system preparations. Bars represent the mean ± SD of the fold change in serum growth factor content of LP-PRP activated with CaCl2 (blue bars; n=5 donors) or exogenously with CaCl2 + thrombin (red bars; n=7 donors). *p<0.03 compared with whole blood. **p<0.01 compared with whole blood. The green line represents whole blood platelet growth factor levels (set to 1-fold for comparison). LP-PRP, leucocyte-poor platelet-rich plasma.

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