Cellular Uptake and Intracellular Trafficking of Poly(N-(2-Hydroxypropyl) Methacrylamide)

被引：6

作者：

Battistella, Claudia ^{[1
,2
]}

Guiet, Romain ^{[3
]}

Burri, Olivier ^{[3
]}

Seitz, Arne ^{[3
]}

Escrig, Stephane ^{[4
]}

Knott, Graham W. ^{[5
]}

Meibom, Anders ^{[4
,6
]}

Klok, Harm-Anton ^{[1
,2
]}

机构：

[1] Ecole Polytech Fed Lausanne, Inst Mat, Batiment MXD,Stn 12, CH-1015 Lausanne, Switzerland

[2] Ecole Polytech Fed Lausanne, Inst Sci & Ingn Chim, Lab Polymeres, Batiment MXD,Stn 12, CH-1015 Lausanne, Switzerland

[3] Ecole Polytech Fed Lausanne, Bioimaging & Opt Platform, Fac Sci Vie, Batiment AI,Stn 15, CH-1015 Lausanne, Switzerland

[4] Ecole Polytech Fed Lausanne, Sch Architecture Civil & Environm Engn, Lab Biol Geochem, CH-1015 Lausanne, Switzerland

[5] Ecole Polytech Fed Lausanne, Bioelectron Microscopy Core Facil, Fac Sci Vie, Batiment AI,Stn 19, CH-1015 Lausanne, Switzerland

[6] Univ Lausanne, Inst Earth Sci, Ctr Adv Surface Anal, CH-1015 Lausanne, Switzerland

来源：

BIOMACROMOLECULES | 2019年 / 20卷 / 01期

基金：

瑞士国家科学基金会;

关键词：

HPMA COPOLYMERS; DRUG-DELIVERY; COLOCALIZATION; NANOPARTICLES; CELLS; INTERNALIZATION; LOCALIZATION; FLUORINE; ORIGINS; FUTURE;

D O I：

10.1021/acs.biomac.8b01372

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

Cellular uptake and intracellular trafficking of polymer conjugates or polymer nanoparticles is typically monitored using fluorescence-based techniques such as confocal microscopy. While these methods have provided a wealth of insight into the internalization and trafficking of polymers and polymer nanoparticles, they require fluorescent labeling of the polymer or polymer nanoparticle. Because in biological media fluorescent dyes may degrade, be cleaved from the polymer or particle, or even change uptake and trafficking pathways, there is an interest in fluorescent label-free methods to study the interactions between cells and polymer nanomedicines. This article presents a first proof-of-concept that demonstrates the feasibility of NanoSIMS to monitor the intracellular localization of polymer conjugates. For the experiments reported here, poly(N-(2-hydroxypropyl) methacrylamide)) (PHPMA) was selected as a prototypical polymer-drug conjugate. This PHPMA polymer contained a F-19-label at the alpha-terminus, which was introduced in order to allow NanoSIMS analysis. Prior to the NanoSIMS experiments, the uptake and intracellular trafficking of the polymer was established using confocal microscopy and flow cytometry. These experiments not only provided detailed insight into the kinetics of these processes but were also important to select time points for the NanoSIMS analysis. For the NanoSIMS experiments, HeLa cells were investigated that had been exposed to the PHPMA polymer for a period of 4 or 15 h, which was known to lead to predominant lysosomal accumulation of the polymer. NanoSIMS analysis of resin-embedded and microtomed samples of the cells revealed a punctuated fluorine signal, which was found to colocalize with the sulfur signal that was attributed to the lysosomal compartments. The localization of the polymer in the endolysosomal compartments was confirmed by TEM analysis on the same cell samples. The results of this study illustrate the potential of NanoSIMS to study the uptake and intracellular trafficking of polymer nanomedicines.

引用

页码：231 / 242

页数：12

共 46 条

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