Pluronic decorated-nanogels with temperature-responsive volume transitions, cytotoxicities, and transfection efficiencies
Pluronic decorated-nanogels with temperature-responsive volume transitions, cytotoxicities, and transfection efficiencies
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DOI：
10.1016/j.ejpb.2008.05.027
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发表时间：
2008-10-01
影响因子：
4.9
通讯作者：
Yoo, Hyuk Sang
中科院分区：
文献类型：
--
作者：
Lee, Jung Im;Yoo, Hyuk Sang
DNA纳米凝胶是通过将Pluronic化学缀合到阳离子聚合物/DNA复合物的表面来制备的，以制备具有内体破坏能力的热响应性纳米凝胶。胺反应性 Pluronic 通过激活 Pluronic 的羟基而制备，随后与预先形成的 PEI/DNA 复合物反应。通过测量缀合反应期间释放的硝基苯基来监测缀合过程。当温度从 20 摄氏度升高到 37 摄氏度时，纳米凝胶的特性（尺寸和 zeta 电位）发生显着变化。纳米凝胶的多峰尺寸分布也证实了纳米凝胶在温度变化时的可变尺寸和分布。电子显微镜和原子力显微镜也证实了热响应纳米凝胶的形态和尺寸的调制。共聚焦显微镜显示纳米凝胶在低温下破坏溶酶体和内体，从而证实了内体破坏能力。存活率结果表明，随着温度从37℃降低至20℃，纳米凝胶的细胞毒性增加，表明塌陷的Pluronic链在调节细胞毒性中发挥作用。还在 NIH3T3 细胞中测量了纳米凝胶的体外转染效率。随着温度降低至 20°C，转染效率增加，从而证实内体破坏在提高转染效率方面发挥了重要作用。 (C) 2008 Elsevier B.V. 保留所有权利。
DNA nanogels were prepared by chemically conjugating Pluronic to the surface of cationic polymer/DNA complex in order to prepare thermo-responsive nanogels with endosomal disrupting abilities. Amine-reactive Pluronic was prepared by activating hydroxyl groups of Pluronic and subsequently reacted with pre-formed PEI/DNA complex. The conjugation process was monitored by measuring liberated nitrophenyl groups during the conjugation reaction. The properties of the nanogels (size and zeta-potential) changed significantly when temperature was increased from 20 to 37 degrees C. The multimodal size distribution of the nanogel also confirmed the variable sizes and distribution of the nanogel upon changing temperatures. Electron microscopy and atomic force microscopy also confirmed the modulated morphologies and sizes of the thermo-responsive nanogel. Confocal microscopy revealed that the nanogel disrupted lysosomes and endosomes at low temperatures, thus confirming endosomal disrupting abilities. The survival rates results showed that the cytotoxicities of the nanogel increased as the temperature decreased from 37 to 20 degrees C, showing that collapsed Pluronic chain played a role in modulating cytotoxicities. In vitro transfection efficiencies of the nanogel were also measured in NIH3T3 cells. Transfection efficiencies increased as the temperature decreased to 20 degrees C, thus confirming that endosomal disruptions played a significant role in increasing transfection efficiencies. (C) 2008 Elsevier B.V. All rights reserved.