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用于快速简便分离尿细胞外囊泡的嵌合纳米复合材料。

Chimeric nanocomposites for the rapid and simple isolation of urinary extracellular vesicles.

机构信息

Department of Biotechnology, College of Life Science and Biotechnology, Yonsei University, Seoul, Republic of Korea.

Department of Urology, Asan Medical Center, University of Ulsan College of Medicine, Seoul, Republic of Korea.

出版信息

J Extracell Vesicles. 2022 Feb;11(2):e12195. doi: 10.1002/jev2.12195.

Abstract

Cancer cell-derived extracellular vesicles (EVs) are promising biomarkers for cancer diagnosis and prognosis. However, the lack of rapid and sensitive isolation techniques to obtain EVs from clinical samples at a sufficiently high yield limits their practicability. Chimeric nanocomposites of lactoferrin conjugated 2,2-bis(methylol)propionic acid dendrimer-modified magnetic nanoparticles (LF-bis-MPA-MNPs) are fabricated and used for simple and sensitive EV isolation from various biological samples via a combination of electrostatic interaction, physically absorption, and biorecognition between the surfaces of the EVs and the LF-bis-MPA-MNPs. The speed, efficiency, recovery rate, and purity of EV isolation by the LF-bis-MPA-MNPs are superior to those obtained by using established methods. The relative expressions of exosomal microRNAs (miRNAs) from isolated EVs in cancerous cell-derived exosomes are verified as significantly higher than those from noncancerous ones. Finally, the chimeric nanocomposites are used to assess urinary exosomal miRNAs from urine specimens from 20 prostate cancer (PCa), 10 benign prostatic hyperplasia (BPH), patients and 10 healthy controls. Significant up-regulation of miR-21 and miR-346 and down-regulation of miR-23a and miR-122-5p occurs in both groups compared to healthy controls. LF-bis-MPA-MNPs provide a rapid, simple, and high yield method for human excreta analysis in clinical applications.

摘要

癌细胞衍生的细胞外囊泡(EVs)是癌症诊断和预后的有前途的生物标志物。然而,缺乏快速和敏感的分离技术,无法从临床样本中以足够高的产量获得 EV,限制了它们的实用性。本文制备了乳铁蛋白偶联 2,2-双(羟甲基)丙酸树枝状大分子修饰的磁性纳米粒子(LF-bis-MPA-MNPs)的嵌合纳米复合材料,并通过 EV 与 LF-bis-MPA-MNPs 表面之间的静电相互作用、物理吸附和生物识别的结合,从各种生物样本中简单、灵敏地分离 EV。LF-bis-MPA-MNPs 分离 EV 的速度、效率、回收率和纯度均优于使用现有方法获得的结果。从分离的 EV 中获得的癌源性细胞衍生的外泌体中的外泌体 microRNAs(miRNAs)的相对表达明显高于非癌性细胞衍生的外泌体中的相对表达。最后,使用嵌合纳米复合材料评估了来自 20 例前列腺癌(PCa)、10 例良性前列腺增生(BPH)患者和 10 例健康对照者尿液标本中的尿外泌体 miRNAs。与健康对照组相比,两组的 miR-21 和 miR-346 均显著上调,miR-23a 和 miR-122-5p 均显著下调。LF-bis-MPA-MNPs 为临床应用中人类排泄物的分析提供了一种快速、简单、高产的方法。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5f28/8859916/5187e1c49da4/JEV2-11-e12195-g007.jpg

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