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一种新型硅基平台,用于选择性分离、定量和分子分析小型细胞外囊泡。

A Novel Silicon Platform for Selective Isolation, Quantification, and Molecular Analysis of Small Extracellular Vesicles.

机构信息

Dipartimento di Fisica e Astronomia, Università di Catania, Catania, 95123, Italy.

CNR-IPCF, Istituto per i Processi Chimico-Fisici, Messina, 98158, Italy.

出版信息

Int J Nanomedicine. 2021 Sep 28;16:5153-5165. doi: 10.2147/IJN.S310896. eCollection 2021.

Abstract

INTRODUCTION

Small extracellular vesicles (sEVs), thanks to their cargo, are involved in cellular communication and play important roles in cell proliferation, growth, differentiation, apoptosis, stemness and embryo development. Their contribution to human pathology has been widely demonstrated and they are emerging as strategic biomarkers of cancer, neurodegenerative and cardiovascular diseases, and as potential targets for therapeutic intervention. However, the use of sEVs for medical applications is still limited due to the selectivity and sensitivity limits of the commonly applied approaches.

METHODS

Novel sensing solutions based on nanomaterials are arising as strategic tools able to surpass traditional sensor limits. Among these, Si nanowires (Si NWs), realized with cost-effective industrially compatible metal-assisted chemical etching, are perfect candidates for sEV detection.

RESULTS

In this paper, the realization of a selective sensor able to isolate, concentrate and quantify specific vesicle populations, from minimal volumes of biofluid, is presented. In particular, this Si NW platform has a detection limit of about 2×10 sEVs/mL and was tested with follicular fluid and blastocoel samples. Moreover, the possibility to detach the selectively isolated sEVs allowing further analyses with other approaches was demonstrated by SEM analysis and several PCRs performed on the RNA content of the detached sEVs.

DISCUSSION

This platform overcomes the limit of detection of traditional methods and, most importantly, preserves the biological content of sEVs, opening the route toward a reliable liquid biopsy analysis.

摘要

简介

由于其携带的货物,小细胞外囊泡(sEVs)参与细胞通讯,并在细胞增殖、生长、分化、凋亡、干性和胚胎发育中发挥重要作用。它们对人类病理学的贡献已得到广泛证明,并且作为癌症、神经退行性和心血管疾病的战略生物标志物以及治疗干预的潜在靶点而出现。然而,由于常用方法的选择性和灵敏度限制,sEVs 在医学应用中的使用仍然受到限制。

方法

基于纳米材料的新型传感解决方案作为战略工具出现,能够超越传统传感器的限制。在这些解决方案中,通过具有成本效益的工业兼容金属辅助化学蚀刻实现的硅纳米线(Si NWs)是检测 sEV 的理想选择。

结果

在本文中,提出了一种选择性传感器的实现,该传感器能够从最小体积的生物流体中分离、浓缩和定量特定的囊泡群体。特别是,该 Si NW 平台的检测限约为 2×10 sEVs/mL,并已在卵泡液和囊胚样本中进行了测试。此外,通过 SEM 分析和对分离的 sEVs 的 RNA 内容进行的多次 PCR,证明了可以分离选择性分离的 sEVs 以允许使用其他方法进行进一步分析的可能性。

讨论

该平台克服了传统方法的检测极限,最重要的是,保留了 sEVs 的生物内容,为可靠的液体活检分析开辟了道路。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e0bc/8487288/6d35c6a6a891/IJN-16-5153-g0001.jpg

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