Department of Orthopaedic Surgery, Hallym University Kangnam Sacred Heart Hospital, Seoul, South Korea.
Department of Orthopaedic Surgery, Seoul National University College of Medicine, Seoul, South Korea.
J Orthop Res. 2020 Apr;38(4):777-784. doi: 10.1002/jor.24520. Epub 2019 Nov 19.
This study aimed to evaluate growth factor concentration in platelet-rich plasma (PRP) (leukocyte-rich PRP) based on storage temperature, duration of storage, and method of activation. PRP samples were stored at 24℃ (room temperature group), 4℃ (refrigerator group), and -70℃ (deep-freezer group). In each temperature, four aliquots were prepared based on the time of analysis (immediately, 1, 3, and 7 days after preparation). After storage, concentrations of platelet-derived growth factor-AA (PDGF-AA), transforming growth factor-β (TGF-β), vascular endothelial growth factor (VEGF), insulin-like growth factor-1 (IGF-1), and fibroblast growth factor-basic (FGF-B) were assessed with/without activation using Quantikine colorimetric sandwich immunoassay kits. PRP was activated with 10% Triton-X for PDGF-AA, VEGF, FGF-B, IGF-1 measurement and sonication for TGF-β1 measurement. Without activation, PDGF-AA concentration was highest on day 7 in the room temperature group. With activation, the concentration of PDGF-AA was constant over the observation period at all temperatures. Without activation, the TGF-β1 concentration remained negligible over the observation period at all temperatures. However, with activation, TGF-β1 gradually increased to its highest concentration on day 7 at all temperatures. Over the observation period, VEGF and IGF-1 concentrations were constant with and without activation at all temperatures. Without activation, FGF-B concentration increased, with the highest concentration observed on day 7 in the deep-freezer group. With activation, FGF-B concentration decreased after day 1 in the room temperature group. Growth factor concentration in PRP differed significantly based on storage temperature, duration of storage, and method of activation. Appropriate storage conditions and activation are important to optimize its effects on desired clinical outcomes. © 2019 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 38:777-784, 2020.
本研究旨在评估基于储存温度、储存时间和激活方法的富血小板血浆(PRP)(富含白细胞的 PRP)中的生长因子浓度。PRP 样本分别在 24℃(室温组)、4℃(冰箱组)和-70℃(深冻组)下储存。在每个温度下,根据分析时间(制备后立即、1 天、3 天和 7 天)制备 4 个等分试样。储存后,使用 Quantikine 比色夹心免疫测定试剂盒评估血小板衍生生长因子-AA(PDGF-AA)、转化生长因子-β(TGF-β)、血管内皮生长因子(VEGF)、胰岛素样生长因子-1(IGF-1)和碱性成纤维细胞生长因子-basic(FGF-B)的浓度,同时/不进行激活。用 10%Triton-X 激活 PDGF-AA、VEGF、FGF-B 和 IGF-1 的测量,用超声处理激活 TGF-β1 的测量。未经激活,室温组 PRP 中 PDGF-AA 的浓度在第 7 天最高。经激活后,在所有温度下,PDGF-AA 的浓度在整个观察期内保持不变。未经激活,在所有温度下,TGF-β1 的浓度在整个观察期内保持微不足道。然而,经激活后,TGF-β1 在所有温度下的浓度逐渐增加至第 7 天的最高浓度。在整个观察期内,在所有温度下,经激活和未经激活,VEGF 和 IGF-1 的浓度保持不变。未经激活,FGF-B 浓度增加,在深冻组中第 7 天观察到最高浓度。经激活后,室温组中 PDGF-AA 的浓度在第 1 天之后降低。PRP 中的生长因子浓度因储存温度、储存时间和激活方法的不同而有显著差异。适当的储存条件和激活对于优化其对预期临床结果的影响非常重要。