Fluorescence and electron microscopy probes for cellular and tissue uptake of poly(D,L-lactide-co-glycolide) nanoparticles
Fluorescence and electron microscopy probes for cellular and tissue uptake of poly(D,L-lactide-co-glycolide) nanoparticles
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DOI：
10.1016/s0378-5173(03)00295-3
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发表时间：
2003-08-27
影响因子：
5.8
通讯作者：
Labhasetwar, V
中科院分区：
文献类型：
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作者：
Panyam, J;Sahoo, SK;Labhasetwar, V
由聚（D,L-丙交酯-乙交酯）（PLGA）和聚（丙交酯）（PLA）配制的纳米颗粒正在被广泛研究用于不同的治疗应用，例如持续药物、疫苗和基因递送。对于许多此类应用，有必要研究纳米颗粒的细胞内分布以及组织摄取，以优化封装治疗剂的功效。荧光和电子显微镜技术通常用于上述目的。通常分别用作电子显微镜和荧光显微镜的模型颗粒的胶体金颗粒和荧光聚苯乙烯可能不是这些研究中 PLGA/PLA 纳米颗粒的合适替代品，主要是因为它们的物理性质存在差异，而且它们不含任何治疗剂。本研究的目的是开发和表征适用于共焦/荧光和透射电子显微镜 (TEM) 研究的 PLGA 纳米颗粒配方。为了实现这一目标，配制了含有 6-香豆素作为荧光标记、四氧化锇作为电子显微镜标记、牛血清白蛋白 (BSA) 作为模型蛋白的 PLGA 纳米颗粒。将负载标记的纳米粒子的不同物理性质（例如粒径、zeta电位、残留PVA含量和蛋白质负载）与未负载的纳米粒子进行比较，发现没有显着差异。此外，加载标记的纳米颗粒制剂与未加载的纳米颗粒一样对细胞无毒。负载 6-香豆素的纳米颗粒被发现可用于使用共焦显微镜研究细胞内纳米颗粒的摄取和分布，而负载四氧化锇的纳米颗粒被发现可用于使用 TEM 研究纳米颗粒在细胞和组织中的摄取和分布。结论是，6-香豆素和四氧化锇可以分别作为有用的荧光和电子显微镜探针，用于掺入纳米颗粒中以研究其细胞和组织分布。 (C) 2003 Elsevier B.V. 保留所有权利。
Nanoparticles formulated from Poly(D,L-lactide-co-glycolide) (PLGA) and poly(lactide) (PLA) are being extensively investigated for different therapeutic applications such as for sustained drug, vaccine, and gene delivery. For many of these applications, it is necessary to study the intracellular distribution as well as the tissue uptake of nanoparticles to optimize the efficacy of the encapsulated therapeutic agent. Fluorescence and electron microscopic techniques are usually used for the above purposes. Colloidal gold particles and fluorescent polystyrene, which are generally used as model particles for electron and fluorescence microscopy, respectively, may not be suitable alternatives to PLGA/PLA nanoparticles for these studies mainly because of the differences in their physical properties and also because they do not contain any therapeutic agent. The aim of the present study was to develop and characterize PLGA nanoparticle formulations that would be suitable for confocal/fluorescence and transmission electron microscopic (TEM) Studies. Towards this objective, PLGA nanoparticles containing 6-coumarin as a fluorescent marker and osmium tetroxide as an electron microscopic marker with bovine serum albumin (BSA) as a model protein were formulated. Different physical properties of marker-loaded nanoparticles such as particle size, zeta potential, residual PVA content and protein-loading were compared with those of unloaded nanoparticles and were found to be not significantly different. Furthermore, marker-loaded nanoparticle formulations were non-toxic to the cells as unloaded nanoparticles. Nanoparticles loaded with 6-coumarin were found to be useful for studying intracellular nanoparticle uptake and distribution using confocal microscopy while osmium tetroxide-loaded nanoparticles were found to be useful for studying nanoparticle uptake and distribution in cells and tissue using TEM. It was concluded that 6-coumarin and osmium tetroxide could serve as useful fluorescence and electron microscopy probes, respectively, for incorporation into nanoparticles to study their cellular and tissue distribution. (C) 2003 Elsevier B.V. All rights reserved.