Characterization of the transgene expression generated by branched and linear polyethylenimine-plasmid DNA nanoparticles in vitro and after intraperitoneal injection in vivo
Characterization of the transgene expression generated by branched and linear polyethylenimine-plasmid DNA nanoparticles in vitro and after intraperitoneal injection in vivo
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DOI：
10.1016/j.jconrel.2008.04.014
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发表时间：
2008-09-10
影响因子：
10.8
通讯作者：
Salem, Aliasger K.
中科院分区：
文献类型：
--
作者：
Intra, Janjira;Salem, Aliasger K.
聚乙烯亚胺 (PEI) 是一种阳离子聚合物，已显示出在体外和体内传递基因的巨大潜力。将阳离子 PEI 与带负电荷的质粒 DNA (pDNA) 混合，会自发静电形成稳定的纳米颗粒复合物。 PEI的结构可以是支链的或直链的。在这项研究中，我们发现支链 PEI 与 pDNA 的静电相互作用比线性 PEI 更强，这导致在相同的 pDNA 浓度下具有更大的压实度、更高的 zeta 电位和更小的纳米颗粒尺寸。对于直链和支链 PEI，增加相同体积和相同氮磷 (N:P) 比例混合的 pDNA 浓度会导致平均粒径更大。增加 N:P 比率会增加 HEK293、COS7 和 HeLa 细胞系中分支 PEI-pDNA 纳米颗粒和线性 PEI-pDNA 纳米颗粒产生的荧光素酶活性。增加制备支化 PEI-pDNA 纳米颗粒时的 N:P 比例也会增加 HepG2 细胞中的荧光素酶表达，但不会增加线性 PEI-pDNA 纳米颗粒产生的荧光素酶表达。在所有细胞系中，N:P 比例为 10 及以上制备的支化 PEI-pDNA 纳米颗粒比线性 PEI-pDNA 纳米颗粒产生显着更高的荧光素酶活性。 HEK293细胞中荧光素酶活性最高，各细胞系中荧光素酶表达量遵循HEK293>COS7>HepG2>HeLa的顺序。腹膜内 (19) 注射 PEI-pDNA 纳米颗粒很有吸引力，因为它简单、可重复，并且通常会导致驻留在腹膜中的纳米颗粒复合物产生储库效应。 IP给药途径避免了PEI-pDNA纳米颗粒在肺部积聚，并且纳米颗粒不穿过血脑屏障。在本研究中，利用生物发光成像（BLI），我们表明改变PEI结构和PEI-pDNA纳米颗粒的剂量对体内IP注射后转基因表达的强度和持续时间有显着影响，但增加N:P比率则不会。增加腹膜内注射的所有 PEI-pDNA 纳米颗粒制剂的剂量和 N:P 比率不会降低小鼠的存活率，并且根据身体状况评分 (BCS) 技术确定，所有小鼠仍保持良好的健康状况。 (C) 2008 Elsevier B.V. 保留所有权利。
Polyethylenimine (PEI) is a cationic polymer that has shown significant potential for delivering genes in vitro and in vivo. Mixing cationic PEI with negatively charged plasmid DNA (pDNA) results in the spontaneous electrostatic formation of stable nanoparticle complexes. The structure of PEI can be branched or linear. In this study, we show that branched PEI has a stronger electrostatic interaction with pDNA than linear PEI, which accounts for greater compaction, higher zeta potentials and smaller nanoparticle sizes at equivalent pDNA concentrations. For both linear and branched PEI, increasing the concentration of pDNA mixed in the same volume and at the same nitrogen to phosphate (N:P) ratio results in larger average particle sizes. Increasing the N:P ratio increases luciferase activity generated by branched PEI-pDNA nanoparticles and linear PEI-pDNA narroparticles in HEK293, COS7 and HeLa cell lines. Increasing the N:P ratio at which branched PEI-pDNA nanoparticles are prepared also increases luciferase expression in HepG2 cells but does not increase luciferase expression generated by linear PEI-pDNA nanoparticles. In all of the cell lines, branched PEI-pDNA nanoparticles prepared at N:P ratios of 10 and above generated significantly higher luciferase activity than linear PEI-pDNA nanoparticles. Luciferase activity was highest in the HEK293 cells and luciferase expression in each of the cell lines followed the order of HEK293>COS7>HepG2>HeLa. Intraperitoneal (19) injection of PEI-pDNA nanoparticles is attractive because it is simple, reproducible and often leads to a depot effect of nanoparticle complexes residing in the peritoneum. The IP route of administration avoids PEI-pDNA nanoparticle accumulation in the lung and the nanoparticles do not pass through the blood-brain barrier.In this study, using bioluminescent imaging (BLI), we show that changing the PEI structure and dose of the PEI-pDNA nanoparticles has a significant impact on the strength and duration of transgene expression after IP injection in vivo but increasing the N:P ratio does not. Increasing the dose and N:P ratio for all the PEI-pDNA nanoparticle formulations injected IP did not reduce mice survival and all mice remained in good health as determined by the Body Condition Scoring (BCS) technique. (C) 2008 Elsevier B.V. All rights reserved.