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富血小板血浆粉末:一种用于生长因子浓度标准化的新制备方法。

Platelet-Rich Plasma Powder: A New Preparation Method for the Standardization of Growth Factor Concentrations.

作者信息

Kieb Matthias, Sander Frank, Prinz Cornelia, Adam Stefanie, Mau-Möller Anett, Bader Rainer, Peters Kirsten, Tischer Thomas

机构信息

Department of Orthopedics, Rostock University Medical Center, Rostock, Germany.

Center of Surgery, Ernst von Bergmann Hospital, Potsdam, Germany.

出版信息

Am J Sports Med. 2017 Mar;45(4):954-960. doi: 10.1177/0363546516674475. Epub 2016 Dec 13.

Abstract

BACKGROUND

Platelet-rich plasma (PRP) is widely used in sports medicine. Available PRP preparations differ in white blood cell, platelet, and growth factor concentrations, making standardized research and clinical application challenging.

PURPOSE

To characterize a newly standardized procedure for pooled PRP that provides defined growth factor concentrations.

STUDY DESIGN

Controlled laboratory study.

METHODS

A standardized growth factor preparation (lyophilized PRP powder) was prepared using 12 pooled platelet concentrates (PCs) derived from different donors via apheresis. Blood samples and commercially available PRP (SmartPrep-2) served as controls (n = 5). Baseline blood counts were analyzed. Additionally, single PCs (n = 5) were produced by standard platelet apheresis. The concentrations of vascular endothelial growth factor (VEGF), basic fibroblast growth factor (bFGF), platelet-derived growth factor AB (PDGF-AB), transforming growth factor β1 (TGF-β1), insulin-like growth factor 1 (IGF-1), interleukin (IL)-1α, IL-1β, and IL-1 receptor agonist (IL-1RA) were analyzed by enzyme-linked immunosorbent assay, and statistical analyses were performed using descriptive statistics, mean differences, 95% CIs, and P values (analysis of variance).

RESULTS

All growth factor preparation methods showed elevated concentrations of the growth factors VEGF, bFGF, PDGF-AB, and TGF-β1 compared with those of whole blood. Large interindividual differences were found in VEGF and bFGF concentrations. Respective values (mean ± SD in pg/mL) for whole blood, SmartPrep-2, PC, and PRP powder were as follows: VEGF (574 ± 147, 528 ± 233, 1087 ± 535, and 1722), bFGF (198 ± 164, 410 ± 259, 151 ± 99, and 542), PDGF-AB (2394 ± 451, 17,846 ± 3087, 18,461 ± 4455, and 23,023), and TGF-β1 (14,356 ± 4527, 77,533 ± 13,918, 68,582 ± 7388, and 87,495). IGF-1 was found in SmartPrep-2 (1539 ± 348 pg/mL). For PC (2266 ± 485 pg/mL), IGF-1 was measured at the same levels of whole blood (2317 ± 711 pg/mL) but was not detectable in PRP powder. IL-1α was detectable in whole blood (111 ± 35 pg/mL) and SmartPrep-2 (119 ± 44 pg/mL).

CONCLUSION

Problems with PRP such as absent standardization, lack of consistency among studies, and black box dosage could be solved by using characterized PRP powder made by pooling and lyophilizing multiple PCs. The new PRP powder opens up new possibilities for PRP research as well as for the treatment of patients.

CLINICAL RELEVANCE

The preparation of pooled PRP by means of lyophilization may allow physicians to apply a defined amount of growth factors by using a defined amount of PRP powder. Moreover, PRP powder as a dry substance with no need for centrifugation could become ubiquitously available, thus saving time and staff resources in clinical practice. However, before transferring the results of this basic science study to clinical application, regulatory issues have to be cleared.

摘要

背景

富血小板血浆(PRP)在运动医学中被广泛应用。现有的PRP制剂在白细胞、血小板和生长因子浓度方面存在差异,这使得标准化研究和临床应用具有挑战性。

目的

对一种新的标准化混合PRP程序进行特性分析,该程序可提供确定的生长因子浓度。

研究设计

对照实验室研究。

方法

使用通过单采术从不同供体获得的12份混合血小板浓缩物(PC)制备标准化生长因子制剂(冻干PRP粉末)。血液样本和市售PRP(SmartPrep-2)作为对照(n = 5)。分析基线血细胞计数。此外,通过标准血小板单采术制备单个PC(n = 5)。采用酶联免疫吸附测定法分析血管内皮生长因子(VEGF)、碱性成纤维细胞生长因子(bFGF)、血小板衍生生长因子AB(PDGF-AB)、转化生长因子β1(TGF-β1)、胰岛素样生长因子1(IGF-1)、白细胞介素(IL)-1α、IL-1β和IL-1受体拮抗剂(IL-1RA)的浓度,并使用描述性统计、均值差异、95%置信区间和P值(方差分析)进行统计分析。

结果

与全血相比,所有生长因子制备方法均显示VEGF、bFGF、PDGF-AB和TGF-β1生长因子浓度升高。发现VEGF和bFGF浓度存在较大个体间差异。全血、SmartPrep-2、PC和PRP粉末的各自值(以pg/mL为单位的均值±标准差)如下:VEGF(574±147、528±233、1087±535和1722)、bFGF(198±164、410±259、151±99和542)、PDGF-AB(2394±451、17846±3087、18461±4455和23023)以及TGF-β1(14356±4527、77533±13918、68582±7388和87495)。在SmartPrep-2中发现了IGF-1(1539±348 pg/mL)。对于PC(2266±485 pg/mL),IGF-1的测量水平与全血(2317±711 pg/mL)相同,但在PRP粉末中未检测到。IL-1α在全血(111±35 pg/mL)和SmartPrep-2(119±44 pg/mL)中可检测到。

结论

通过混合和冻干多个PC制成的特性化PRP粉末可以解决PRP存在的标准化问题、研究之间缺乏一致性以及黑箱剂量等问题。新的PRP粉末为PRP研究以及患者治疗开辟了新的可能性。

临床意义

通过冻干制备混合PRP可能使医生能够通过使用确定量的PRP粉末来应用确定量的生长因子。此外,PRP粉末作为一种无需离心的干燥物质可以随处可得,从而在临床实践中节省时间和人力资源。然而,在将这项基础科学研究的结果转化为临床应用之前,必须解决监管问题。

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