Colloidally stable novel copolymeric system for gene delivery in complete growth media
Colloidally stable novel copolymeric system for gene delivery in complete growth media
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DOI：
10.1016/j.jconrel.2007.05.008
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发表时间：
2007-08-16
影响因子：
10.8
通讯作者：
Mallapragada, Surya K.
中科院分区：
文献类型：
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作者：
Agarwal, Ankit;Vilensky, Rita;Mallapragada, Surya K.
基于聚（2-二乙基氨基乙基甲基丙烯酸酯）（PDEAEM）和普朗尼克F127的新型阳离子五嵌段共聚物作为非病毒基因递送载体进行了评价，从生理化学的角度来看，在完全生长培养基中的稳定性和转染效率。引入了一种新的策略来空间稳定这种基于Pluronic的阳离子聚合物的聚合复合物，以防止与血清蛋白聚集。当阳离子五嵌段共聚物将质粒DNA缩合成100-150 nm直径的纳米复合物时，添加到制剂中的未改性Pluronic与聚合复合物表面上的五嵌段共聚物自组装，并用其长聚（环氧乙烷）链在空间上屏蔽五嵌段共聚物的阳离子PDEAEM链。这些包被的复合物在补充血清的缓冲液中形成150-250 nm直径的胶体稳定分散体。冷冻-TEM显微照片还表明，未改性的普朗尼克涂层复合物减少血清蛋白的聚集。五嵌段共聚物保留了聚合复合物内凝聚的质粒DNA的完整性，并对其被核酸酶降解提供了有效的抗性。虽然在核酸酶消化后由ExGen 500（（R））聚合复合物保留的DNA的总量多于由五嵌段共聚物保留的DNA的总量，但以超螺旋形式保留的质粒的量没有显著差异。用未修饰的Pluronic包被的聚合复合物在完全生长培养基中的SKOV 3细胞中提供了有效的转染，在转染的细胞数量方面与ExGen 5000提供的相当，并且在表达的总转基因蛋白方面低一个数量级。这些空间屏蔽的聚合复合物还表现出比未涂覆的五嵌段共聚物的聚合复合物低得多的细胞毒性，并且显著低于相关浓度下ExGen 500（（R））的细胞毒性。这种胶体稳定的、多功能的、多组分的基因递送系统还在生理温度下形成热可逆的可注射水凝胶，如Pluronics，其可用于聚合复合物的持续递送，并且有希望用于全身应用。(C)2007 Elsevier B. V.保留所有权利。
Novel cationic pentablock copolymers based on poly(2-diethylaminoethylmethacrylate) (PDEAEM) and Pluronic F127 were evaluated as non-viral gene delivery vectors from a physiochemical point of view for stability and transfection efficiency in complete growth media. A novel strategy was introduced to sterically stabilize the polyplexes of such Pluronic-based cationic polymers against aggregation with serum proteins. As cationic pentablock copolymers condense plasmid DNA into nanoplexes of 100-150 nm diameter, unmodified Pluronic added to the formulation self-assemble with the pentablock copolymers on the surface of polyplexes and shield the cationic PDEAEM chains of pentablock copolymers sterically with its long poly(ethyleneoxide) chains. These coated polyplexes formed colloidally stable dispersions of 150-250 nm diameter in serum-supplemented buffers. Cryo-TEM micrographs also showed that coating polyplexes with unmodified Pluronic reduced aggregation in serum proteins. Pentablock copolymers preserved the integrity of plasmid DNA condensed inside the polyplexes and provided efficient resistance to its degradation by nucleases. Though the total amount of DNA retained by ExGen 500((R)) polyplexes after nuclease digestion was more than that retained by pentablock copolymers, the amount of plasmid retained in supercoiled form was not significantly different. Polyplexes coated with unmodified Pluronic provided efficient transfection in SKOV3 cells in complete growth media, comparable to that provided by ExGen 5000 in terms of number of cells transfected, and one order less in terms of total transgene protein expressed. These sterically shielded polyplexes also exhibited much lower cytotoxicities than uncoated polyplexes of pentablock copolymers, and significantly lower than the cytotoxicity of ExGen 500((R)) at relevant concentrations. This colloidally stable, versatile, multi-component gene delivery system also forms thermo-reversible injectable hydrogels like Pluronics at physiological temperatures that can be used for sustained delivery of polyplexes, and is promising for systemic applications. (C) 2007 Elsevier B.V. All rights reserved.