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少白细胞富血小板血浆制备的优化:对使用不同制备、储存和激活方法获得的少白细胞富血小板血浆的验证研究

Optimization of leukocyte-poor platelet-rich plasma preparation: a validation study of leukocyte-poor platelet-rich plasma obtained using different preparer, storage, and activation methods.

作者信息

Kikuchi Naoya, Yoshioka Tomokazu, Taniguchi Yu, Sugaya Hisashi, Arai Norihito, Kanamori Akihiro, Yamazaki Masashi

机构信息

Department of Orthopedic Surgery, Faculty of Medicine, University of Tsukuba, 1-1-1 Tennodai, Tsukuba, Ibaraki, 305-8575, Japan.

Regenerative Medicine for Musculoskeletal System, Faculty of Medicine, University of Tsukuba, 1-1-1 Tennodai, Tsukuba, Ibaraki, 305-8575, Japan.

出版信息

J Exp Orthop. 2019 Jun 3;6(1):24. doi: 10.1186/s40634-019-0190-8.

Abstract

BACKGROUND

Alternative methods of platelet-rich plasma (PRP) preparation, storage, and activation that can be stably reproduced are needed to improve PRP production. The purpose of this study was to investigate the effect of the preparer's experience on the quality of prepared PRP, chronological changes occurring in PRP, and the effect of the activation procedures on the release of several growth factors from PRP, using PRP prepared with the PRGF-Endoret Kit.

METHODS

Leukocyte-poor PRP samples from seventeen healthy volunteers were prepared using the PRGF-Endoret Kit and the PRGF IV System Centrifuge. The platelet and leukocyte concentrations were compared based on the preparer's experience. The concentrations of platelets, hepatocyte growth factor (HGF), platelet-derived growth factor-BB (PDGF-BB), and insulin-like growth factor-1 (IGF-1) were determined at 0 and 60 min after PRP preparation, and compared. Concentrations of the above growth factors from PRP activated by freeze-thaw cycling and by calcium chloride (CaCl) were also compared.

RESULTS

No significant difference was observed in the platelet concentrations and leukocyte contamination rates, based on the preparer's experience. At 60 min after PRP preparation, the platelet concentration decreased significantly, while the HGF, PDGF-BB, and IGF-1 concentrations remained unchanged. Activation with CaCl resulted in a significant increase in the PDGF-BB levels, although the HGF and IGF-1 concentrations remained unchanged.

CONCLUSIONS

The results of this study show that leukocyte-poor PRP prepared using the PRGF-Endoret Kit did not result in any qualitative difference that depended on the experience of the preparer. However, PRP preparation required standardization in terms of the time of blood count measurement. Growth factor concentrations in PRP differed according to the platelet-activation method used.

摘要

背景

为了改进富血小板血浆(PRP)的制备,需要可稳定重现的PRP制备、储存和激活的替代方法。本研究的目的是使用PRGF-Endoret试剂盒制备的PRP,研究制备者经验对所制备PRP质量的影响、PRP随时间的变化以及激活程序对PRP中几种生长因子释放的影响。

方法

使用PRGF-Endoret试剂盒和PRGF IV系统离心机,从17名健康志愿者中制备贫白细胞PRP样本。根据制备者的经验比较血小板和白细胞浓度。在PRP制备后0分钟和60分钟测定血小板、肝细胞生长因子(HGF)、血小板衍生生长因子-BB(PDGF-BB)和胰岛素样生长因子-1(IGF-1)的浓度,并进行比较。还比较了通过冻融循环和氯化钙(CaCl)激活的PRP中上述生长因子的浓度。

结果

根据制备者的经验,血小板浓度和白细胞污染率未观察到显著差异。在PRP制备后60分钟,血小板浓度显著下降,而HGF、PDGF-BB和IGF-1浓度保持不变。用CaCl激活导致PDGF-BB水平显著增加,尽管HGF和IGF-1浓度保持不变。

结论

本研究结果表明,使用PRGF-Endoret试剂盒制备的贫白细胞PRP在质量上没有因制备者经验而产生任何差异。然而,PRP制备在血细胞计数测量时间方面需要标准化。PRP中的生长因子浓度因所用的血小板激活方法而异。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8cd4/6546777/3adaed9b29ae/40634_2019_190_Fig1_HTML.jpg

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