Intracellular uptake and trafficking of difluoroboron dibenzoylmethane-polylactide nanoparticles in HeLa cells.
Intracellular uptake and trafficking of difluoroboron dibenzoylmethane-polylactide nanoparticles in HeLa cells.
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DOI：
10.1021/nn901385y
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发表时间：
2010-05-25
期刊：
影响因子：
17.1
通讯作者：
Hamm-Alvarez SF
中科院分区：
文献类型：
--
作者：
Contreras J;Xie J;Chen YJ;Pei H;Zhang G;Fraser CL;Hamm-Alvarez SF
在本研究中，制备了基于二氟硼二苯甲酰甲烷-聚乳酸（BF2dbmPLA）的纳米颗粒。聚乳酸或聚丙交酯是常用的可降解聚合物，而硼染料具有消光系数大、发射量子产率高、2光子吸收和对周围环境敏感等优点。 BF2dbmPLA 表现出分子量依赖性发射特性，并且可以配制为稳定的纳米颗粒，这表明其独特的光学特性可能在多种环境下用于探测细胞内环境。在这里，我们表明 BF2dbmPLA 纳米颗粒通过内吞作用内化到培养的 HeLa 细胞中，并且在细胞环境中它们保留了其荧光特性。 BF2dbmPLA 纳米粒子具有光稳定性，在破坏常见光稳定性探针 LysoTracker™ 蓝色荧光的条件下，可抵抗激光诱导的光漂白。它们的内吞作用也是脂筏依赖性的，摄取后它们与膜区室中霍乱毒素 B 亚基的显着共定位以及它们对甲基-β-环糊精摄取的敏感性证明了这一点。此外，BF2dbmPLA 纳米颗粒内吞作用利用微管和肌动蛋白丝。内化的 BF2dbmPLA 纳米颗粒不会在酸性晚期核内体和溶酶体中积累，而是在核周非溶酶体区室中积累。这些发现证明了使用具有多种发射特性的新型 BF2dbmPLA 纳米颗粒进行原位活细胞成像的可行性，并表明它们的内源摄取是通过脂筏依赖性内吞机制发生的。
In this study, nanoparticles based on difluoroboron dibenzoylmethane-poly(lactic acid) (BF2dbmPLA) are prepared. Polylactic acid or polylactide is a commonly used degradable polymer, while the boron dye possesses a large extinction coefficient, high emission quantum yield, 2-photon absorption, and sensitivity to the surrounding environment. BF2dbmPLA exhibits molecular weight-dependent emission properties, and can be formulated as stable nanoparticles, suggesting that its unique optical properties may be useful in multiple contexts for probing intracellular environments. Here we show that BF2dbmPLA nanoparticles are internalized into cultured HeLa cells by endocytosis, and that within the cellular milieu they retain their fluorescence properties. BF2dbmPLA nanoparticles are photostable, resisting laser-induced photobleaching under conditions that destroy the fluorescence of a common photostable probe, LysoTracker™ blue. Their endocytosis is also lipid raft-dependent, as evidenced by their significant co-localization with cholera toxin B subunit in membrane compartments after uptake, and their sensitivity of uptake to methyl-β-cyclodextrin. Additionally, BF2dbmPLA nanoparticle endocytosis utilizes microtubules and actin filaments. Internalized BF2dbmPLA nanoparticles do not accumulate in acidic late endosomes and lysosomes, but within a perinuclear non-lysosomal compartment. These findings demonstrate the feasibility of using novel BF2dbmPLA nanoparticles exhibiting diverse emission properties for in situ, live cell imaging, and suggest that their endogenous uptake occurs through a lipid-raft dependent endocytosis mechanism.
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影响因子：
4.3
作者：
Cogné-Laage, E;Allemand, JF;Jullien, L
通讯作者：
Jullien, L
影响因子：
13.3
作者：
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通讯作者：
Drezek, Rebekah
影响因子：
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作者：
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Mailaender, Volker
影响因子：
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作者：
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通讯作者：
Kruth, Howard S.
影响因子：
6.2
作者：
Harush-Frenkel, Oshrat;Rozentur, Eva;Altschuler, Yoram
通讯作者：
Altschuler, Yoram