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对在不同离心力(g)和时间下制备的富血小板血浆(PRP)中的血小板及血小板衍生生长因子进行定量分析。

Quantification of platelets and platelet derived growth factors from platelet-rich-plasma (PRP) prepared at different centrifugal force (g) and time.

作者信息

Arora Satyam, Doda Veena, Kotwal Urvershi, Dogra Mitu

机构信息

Department of Transfusion Medicine, Dr Ram Manohar Lohia Hospital, New Delhi, India.

Department of Transfusion Medicine, Dr Ram Manohar Lohia Hospital, New Delhi, India.

出版信息

Transfus Apher Sci. 2016 Feb;54(1):103-10. doi: 10.1016/j.transci.2016.01.028. Epub 2016 Feb 3.

Abstract

INTRODUCTION

Platelet derived biomaterials represent a key source of cytokines and growth factors extensively used for tissue regeneration; wound healing and tissue repair. Our study was to quantify platelets and growth factors released by PRP when prepared at different centrifugal force (g) and time.

MATERIAL AND METHODS

Our study was approved by the institutional ethical committee. One hundred millilitres of whole blood (WB) was collected in bag with CPDA as the anticoagulant(AC); (14 mL for 100 mL WB ratio). Nine aliquots of 10 mL each were made from the bag and set of three aliquots were made a group. PRP was prepared at varying centrifugal force (group A: -110 g, group B: -208 g & group C: -440 g) & time (1: -5 min, 2: -10 min & 3: -20 min). Contents of each PRP prepared were analysed. Commercial sandwich ELISA kits were used to quantify the concentrations of CD62P (Diaclone SAS; France), Platelet derived growth factors-AB (Qayee-Bio; China), transforming growth factor-β1 (DRG; Germany) and vascular endothelial growth factor (Boster Immuno Leader; USA) released in each PRP prepared.

RESULTS

Eight volunteers were enrolled in the study (24-30 years). The baseline blood counts of all the volunteers were comparable (p ≥ 0.05). Mean ± SD of platelet yield of all nine groups ranged from 17.2 ± 4.2% to 78.7 ± 5.7%. Each PRP was activated with calcified thromboplastin to quantify the growth factors released by them. Significantly higher (p < 0.05) transforming growth factor-β1 and vascular endothelial growth factor were released compared to the baseline.

CONCLUSION

Our study highlights the variation in both force (g) and time results in changes at cellular level and growth factor concentrations.

摘要

引言

血小板衍生生物材料是细胞因子和生长因子的重要来源,广泛应用于组织再生、伤口愈合和组织修复。我们的研究旨在量化在不同离心力(g)和时间制备的富血小板血浆(PRP)释放的血小板和生长因子。

材料与方法

我们的研究获得了机构伦理委员会的批准。以CPDA作为抗凝剂(AC),在袋中采集100毫升全血(WB);(14毫升用于100毫升WB比例)。从袋中取出9个10毫升的等分试样,并将3个等分试样分为一组。在不同的离心力(A组:-110 g,B组:-208 g和C组:-440 g)和时间(1:-5分钟,2:-10分钟和3:-20分钟)下制备PRP。分析制备的每种PRP的成分。使用商业夹心ELISA试剂盒量化在制备的每种PRP中释放的CD62P(法国Diaclone SAS)、血小板衍生生长因子-AB(中国Qayee-Bio)、转化生长因子-β1(德国DRG)和血管内皮生长因子(美国Boster Immuno Leader)的浓度。

结果

8名志愿者参与了该研究(24 - 30岁)。所有志愿者的基线血细胞计数具有可比性(p≥0.05)。所有九组的血小板产量平均值±标准差范围为17.2±4.2%至78.7±5.7%。每种PRP用钙化凝血活酶激活以量化它们释放的生长因子。与基线相比,转化生长因子-β1和血管内皮生长因子的释放显著更高(p < 0.05)。

结论

我们的研究强调了力(g)和时间的变化会导致细胞水平和生长因子浓度的变化。

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