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金标记的嵌段共聚物胶束在多个亚细胞位点显示出金聚集体。

Gold-labeled block copolymer micelles reveal gold aggregates at multiple subcellular sites.

作者信息

Soo Patrick Lim, Sidorov Stanislav N, Mui Jeannie, Bronstein Lyudmila M, Vali Hojatollah, Eisenberg Adi, Maysinger Dusica

机构信息

Department of Chemistry, McGill University, 801 Sherbrooke Street W., Montreal, Quebec, Canada.

出版信息

Langmuir. 2007 Apr 24;23(9):4830-6. doi: 10.1021/la063375s. Epub 2007 Mar 29.


DOI:10.1021/la063375s
PMID:17391054
Abstract

There is increasing interest in the usefulness of block copolymer micelles as drug delivery vehicles. However, their subcellular distribution has not been explored extensively, mostly because of the lack of adequately labeled block copolymers. In a previous study, we showed that fluorescently labeled block copolymer micelles entered living cells and co-localized with cytoplasmic organelles selectively labeled with fluorescent dyes. The details of the observed co-localizations were, however, limited by the resolution of the fluorescence approach, which is ca. 500 nm. Using transmission electron microscopy (TEM), we established time- and concentration-dependent subcellular distributions of gold-labeled micelles within human embryonic kidney (HEK 293) cells and human lung carcinoma (A549) cells. Gold particles were incorporated into poly(4-vinylpyridine)-block-poly(ethylene oxide) (P4VP21-b-PEO45) micelles. Data from dynamic light scattering (DLS) and TEM analyses revealed that the sizes of the gold particles ranged from 4 to 8 nm. The cells survived up to 24 h in the presence of low gold-labeled micelle concentrations (0.73 microg/mL), but cell death occurred at higher concentrations (i.e., kidney cells are more susceptible than lung cells). Over 24 h periods of equivalent exposure, lung cells internalized significantly more gold-incorporated micelles than kidney cells. Although micelles were added to the cell culture media as dispersed colloidal particles, the presence of serum in these media caused aggregation. These aggregates occurred mainly close to the cell plasma membrane at early times (5-10 min); however, at later times (24 h) aggregated particles were seen inside endosomes and lysozomes. Thus, gold-incorporated (labeled) micelles can serve as a valuable extension of the fluorescence approach to visualizing the localization of micelles in subcellular compartments, improving the resolution by at least 20-fold.

摘要

人们越来越关注嵌段共聚物胶束作为药物递送载体的效用。然而,它们在亚细胞水平的分布尚未得到广泛研究,主要原因是缺乏标记充分的嵌段共聚物。在之前的一项研究中,我们表明荧光标记的嵌段共聚物胶束能够进入活细胞,并与用荧光染料选择性标记的细胞质细胞器共定位。然而,所观察到的共定位细节受到荧光方法分辨率的限制,该分辨率约为500纳米。利用透射电子显微镜(TEM),我们确定了金标记胶束在人胚肾(HEK 293)细胞和人肺癌(A549)细胞内的时间和浓度依赖性亚细胞分布。金颗粒被掺入聚(4-乙烯基吡啶)-嵌段-聚(环氧乙烷)(P4VP21-b-PEO45)胶束中。动态光散射(DLS)和TEM分析数据显示,金颗粒的大小在4到8纳米之间。在低金标记胶束浓度(0.73微克/毫升)下,细胞在长达24小时内存活,但在较高浓度下会发生细胞死亡(即肾细胞比肺细胞更敏感)。在等效暴露的24小时期间,肺细胞内化的金掺入胶束明显多于肾细胞。尽管胶束作为分散的胶体颗粒添加到细胞培养基中,但这些培养基中血清的存在会导致聚集。这些聚集体主要在早期(5-10分钟)出现在细胞膜附近;然而,在后期(24小时),在内体和溶酶体内可以看到聚集的颗粒。因此,金掺入(标记)胶束可作为荧光方法的有价值扩展,用于可视化胶束在亚细胞区室中的定位,将分辨率提高至少20倍。

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引用本文的文献

[1]
Drug Release by Direct Jump from Poly(ethylene-glycol-b-ε-caprolactone) Nano-Vector to Cell Membrane.

Molecules. 2016-11-30

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