Carvalho Alexandra, Ferreira Ana Filipa, Soares Maria, Santos Susana, Tomé Patrícia, Machado-Simões Juliana, Pais Ana Sofia, Sousa Ana Paula, Paiva Artur, Almeida-Santos Teresa
Reproductive Medicine Unit, Gynecology, Obstetrics, Reproduction and Neonatology Department, Centro Hospitalar e Universitário de Coimbra, 3004-561 Coimbra, Portugal.
CICS-UBI-Health Sciences Research Centre, University of Beira Interior, 6200-506 Covilhã, Portugal.
Bioengineering (Basel). 2024 Feb 23;11(3):209. doi: 10.3390/bioengineering11030209.
Platelet-rich plasma (PRP) has emerged as a promising therapy in regenerative medicine. However, the lack of standardization in PRP preparation protocols presents a challenge in achieving reproducible and accurate results. This study aimed to optimize the PRP preparation protocol by investigating the impact of two different anticoagulants, sodium citrate (SC) and ethylenediaminetetraacetic acid (EDTA), and resuspension media, plasma versus sodium chloride (NaCl). Platelet recovery rates were calculated and compared between groups, in addition to platelet activity and vascular endothelial growth factor (VEGF) released into plasma after PRP activation. The platelet recovery rate was higher with EDTA in comparison to SC (51.04% vs. 29.85%, = 0.005). Platelet activity was also higher, with a higher expression of two platelet antibodies, platelet surface P-Selectin (CD62p) and PAC-1, in the EDTA group. The concentration of VEGF was higher with SC in comparison to EDTA (628.73 vs. 265.44 pg/mL, = 0.013). Platelet recovery rates and VEGF levels were higher in PRP resuspended in plasma when compared to NaCl (61.60% vs. 48.61%, = 0.011 and 363.32 vs. 159.83 pg/mL, = 0.005, respectively). Our study reinforces the superiority of EDTA (as anticoagulant) and plasma (for resuspension) in obtaining a higher platelet recovery and preserving platelet functionality during PRP preparation.
富血小板血浆（PRP）已成为再生医学中一种很有前景的治疗方法。然而，PRP制备方案缺乏标准化给获得可重复且准确的结果带来了挑战。本研究旨在通过研究两种不同抗凝剂柠檬酸钠（SC）和乙二胺四乙酸（EDTA）以及重悬介质（血浆与氯化钠（NaCl））的影响来优化PRP制备方案。计算并比较了各组之间的血小板回收率，以及PRP激活后释放到血浆中的血小板活性和血管内皮生长因子（VEGF）。与SC相比，EDTA的血小板回收率更高（51.04%对29.85%，P = 0.005）。EDTA组的血小板活性也更高，两种血小板抗体血小板表面P-选择素（CD62p）和PAC-1的表达更高。与EDTA相比，SC的VEGF浓度更高（628.73对265.44 pg/mL，P = 0.013）。与NaCl相比，重悬于血浆中的PRP的血小板回收率和VEGF水平更高（分别为61.60%对48.61%，P = 0.011和363.32对159.83 pg/mL，P = 0.005）。我们的研究强化了EDTA（作为抗凝剂）和血浆（用于重悬）在PRP制备过程中获得更高血小板回收率和保持血小板功能方面的优越性。
