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Preparation and Characterization of Cationic PLA-PEG Nanoparticles for Delivery of Plasmid DNA

机译:阳离子PLA-PEG纳米粒子的制备和表征用于质粒DNA的传递

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摘要

The purpose of the present work was to formulate and evaluate cationic poly(lactic acid)-poly(ethylene glycol) (PLA-PEG) nanoparticles as novel non-viral gene delivery nano-device. Cationic PLA-PEG nanoparticles were prepared by nanoprecipitation method. The gene loaded nanoparticles were obtained by incubating the report gene pEGFP with cationic PLA-PEG nanoparticles. The physicochemical properties (e.g., morphology, particle size, surface charge, DNA binding efficiency) and biological properties (e.g., integrity of the released DNA, protection from nuclease degradation, plasma stability, in vitro cytotoxicity, and in vitro transfection ability in Hela cells) of the gene loaded PLA-PEG nanoparticles were evaluated, respectively. The obtained cationic PLA-PEG nanoparticles and gene loaded nanoparticles were both spherical in shape with average particle size of 89.7 and 128.9 nm, polydispersity index of 0.185 and 0.161, zeta potentials of +28.9 and +16.8 mV, respectively. The obtained cationic PLA-PEG nanoparticles with high binding efficiency (>95%) could protect the loaded DNA from the degradation by nuclease and plasma. The nanoparticles displayed sustained-release properties in vitro and the released DNA maintained its structural and functional integrity. It also showed lower cytotoxicity than Lipofectamine 2000 and could successfully transfect gene into Hela cells even in presence of serum. It could be concluded that the established gene loaded cationic PLA-PEG nanoparticles with excellent properties were promising non-viral nano-device, which had potential to make cancer gene therapy achievable.
机译:本工作的目的是配制和评估阳离子聚(乳酸)-聚(乙二醇)(PLA-PEG)纳米粒子作为新型非病毒基因传递纳米装置。通过纳米沉淀法制备了阳离子PLA-PEG纳米颗粒。通过将报告基因pEGFP与阳离子PLA-PEG纳米颗粒孵育,获得了加载基因的纳米颗粒。物理化学特性(例如形态,粒度,表面电荷,DNA结合效率)和生物学特性(例如释放的DNA的完整性,核酸酶降解保护,血浆稳定性,体外细胞毒性和Hela细胞的体外转染能力)分别评估了载有基因的PLA-PEG纳米粒子的)。所得的阳离子PLA-PEG纳米粒子和基因加载的纳米粒子均为球形,平均粒径分别为89.7和128.9 nm,多分散指数分别为0.185和0.161,ζ电位分别为+28.9和+16.8 mV。所获得的具有高结合效率(> 95%)的阳离子PLA-PEG纳米粒子可以保护加载的DNA免受核酸酶和血浆的降解。纳米粒子在体外显示出持续释放的特性,释放的DNA保持其结构和功能的完整性。它也显示出比Lipofectamine 2000更低的细胞毒性,即使在有血清的情况下也可以成功地将基因转染到Hela细胞中。可以得出的结论是,已建立的具有优异性能的载有基因的阳离子PLA-PEG纳米粒子是有前途的非病毒纳米设备,具有使癌症基因治疗可实现的潜力。

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