Investigating the uptake and intracellular fate of pH-sensitive liposomes by flow cytometry and spectral bio-imaging
Investigating the uptake and intracellular fate of pH-sensitive liposomes by flow cytometry and spectral bio-imaging
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DOI：
10.1016/j.jconrel.2005.10.018
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发表时间：
2006-02-21
影响因子：
10.8
通讯作者：
Peschka-Süss, R
中科院分区：
文献类型：
--
作者：
Huth, US;Schubert, R;Peschka-Süss, R
颗粒药物输送系统的细胞内命运取决于初始内在化模式和随后的细胞内贩运。除了网格蛋白介导的内吞作用外，还讨论了不同的细胞摄取机制，包括小窝介导的内吞作用，吞噬作用，大吞噬细胞增多症和不良特征的非clathrin-non-non-non-non-non-caveolae介导的介导的内细胞增多症。此外，还可以包括融合过程。对于有针对性的药物递送，小窝摄取的机制似乎是最有前途的，因为它不会将内部材料转运到可能降解内部材料的内体和溶酶体。因此，对内部化模式的了解是优化药物靶向的第一步。在两种不同的细胞类型中研究了pH敏感脂质体的摄取和细胞内行为。为了进行研究，采用了两种不同的方法：a）使用不同抑制剂的组合选择性地阻断不同的途径。通过流式细胞仪评估这些影响。 b）将荧光标记的标记物（特异于不同的内吞路线）与荧光标记的脂质体一起孵育，随后检查共定位（光谱生物成像）。COS-7和HUVEC通过不同的机制内化的pH-敏感性脂质体。 HUVEC似乎通过依赖性的内吞作用，小窝依赖性内吞作用和巨细胞增多症来内化，而COS-7细胞的行为不同：此处的大型细胞增多症与摄取无关。我们表明，流式细胞仪和光谱生物成像的结合提供了了解初始内在化模式并遵循脂质体的细胞内命运的可能性，这两者都是优化药物靶向系统的先决条件。 （c）2005 Elsevier B.V.保留所有权利。
The intracellular fate of particulate drug delivery systems is determined by the initial mode of internalization and the subsequent intracellular trafficking. Besides clathrin-mediated endocytosis, different cellular uptake mechanisms for internalization are discussed, including caveolae-mediated endocytosis, phagocytosis, macropinocytosis and the badly characterized non-clathrin-non-caveolae-mediated endocytosis. In addition, fusion processes could also be included. For a targeted drug delivery, the mechanism of caveolae uptake seems to be most promising since it does not transport internalized material to endosomes and lysosomes where internalized material might be degraded. The knowledge of the mode of internalization is therefore the first step for optimized drug targeting.The uptake and the intracellular behavior of pH-sensitive liposomes are investigated in two different cell types. For investigation, two different approaches were taken: a) a combination of different inhibitors was used to selectively block different pathways. These effects were evaluated by flow cytometry. b) Fluorescently labeled markers, specific for the different endocytic routes, were incubated with fluorescently labeled liposomes with a subsequent check for co-localization (spectral bio-imaging).COS-7 and HUVEC internalize pH-sensitive liposomes by means of different mechanisms. HUVEC seem to internalize via clathrin-dependent endocytosis, caveolae-dependent endocytosis and macropinocytosis whereas COS-7 cells behave differently: here macropinocytosis is not involved in the uptake. We show that the combination of flow cytometry and spectral bio-imaging offers the possibility to understand the initial mode of internalization and to follow the intracellular fate of liposomes, both of which are a prerequisite for optimizing drug targeting systems. (C) 2005 Elsevier B.V. All rights reserved.