Improved cellular uptake of chitosan-modi fled PLGA nanospheres by A549 cells
Improved cellular uptake of chitosan-modi fled PLGA nanospheres by A549 cells
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DOI：
10.1016/j.ijpharm.2009.07.023
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发表时间：
2009-12-01
影响因子：
5.8
通讯作者：
Kawashima, Yoshiaki
中科院分区：
文献类型：
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作者：
Tahara, Kohei;Sakai, Takeshi;Kawashima, Yoshiaki
作者先前开发了聚(DL-乳酸-乙醇酸共聚)(PLGA)纳米球(NSS)作为肺部给药的纳米颗粒药物载体。本研究表明壳聚糖(CS)修饰的PLGA NSS(CS-PLGA NSS)优先被人肺腺癌细胞(A549)摄取。采用水-油-水乳化溶剂挥发法制备了聚乳酸纳米粒，并对其进行了表面修饰。测定PLGA NS的平均粒径和表面电荷等物理化学参数，以确定影响细胞摄取PLGA NS的参数。荧光分光光度法检测A549细胞摄取能力，激光共聚焦扫描显微镜观察细胞摄取情况。用MTS法比较非系统和CS-PLGA NS系统的体外细胞毒性。细胞对PLGA NS的摄取随着直径减小到亚微米水平和CS介导的表面修饰而增加。细胞对PLGA NS的摄取是能量依赖的，表现为在较低的孵育温度和高渗生长介质中摄取的减少，高渗生长介质被用作笼蛋白包裹的凹坑内吞的抑制剂。A549细胞以能量依赖的方式摄取CS-PLGA NSS，提示胞内蛋白介导的内吞过程。CS-PLGA NS具有较低的细胞毒性，与非PLGA NS相似。(C)2009爱思唯尔B.V.保留所有权利。
The authors have previously developed poly(DL-lactic-co-glycolic acid) (PLGA) nanospheres (NSs) as a nanoparticulate drug carrier for pulmonary administration. The present study demonstrates that chitosan (CS)-modified PLGA NSs (CS-PLGA NSs) are preferentially taken up by human lung adenocarcinoma cells (A549). PLGA NSs prepared using a water-oil-water emulsion solvent evaporation method were surface-modified by adsorption of CS. The physicochemical parameters of PLGA NS, including average size and surface charge, were measured to identify which parameter influenced cellular uptake of PLGA NS. Uptake was confirmed using fluorescence spectrophotometry and was visualized in A549 cells with confocal laser scanning microscopy (CLSM). The cytotoxicities of non- and CS-PLGA NS systems were compared in vitro by MTS assay. Cellular uptake of PLGA NS increased with decreasing diameter to the submicron level and with CS-mediated surface modification. Cellular uptake of PLGA NS was energy dependent, as shown by a reduction in uptake at lower incubation temperatures and in hypertonic growth medium used as an inhibitor of clathrin-coated pit endocytosis. CS-PLGA NSs were taken up by A549 cells in an energy-dependent manner, suggesting a clathrin-mediated endocytic process. CS-PLGA NS demonstrated low cytotoxicity, similar to non-PLGA NS. (C) 2009 Elsevier B.V. All rights reserved.