Crosslinked nanocarriers based upon poly(ethylene imine) for systemic plasmid delivery: In vitro characterization and in vivo studies in mice
Crosslinked nanocarriers based upon poly(ethylene imine) for systemic plasmid delivery: In vitro characterization and in vivo studies in mice
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DOI：
10.1016/j.jconrel.2007.01.007
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发表时间：
2007-04-23
影响因子：
10.8
通讯作者：
Kissel, Thomas
中科院分区：
文献类型：
--
作者：
Neu, Michael;Germershaus, Oliver;Kissel, Thomas
使用低分子量交联剂二硫代双（琥珀酰亚胺基丙酸酯）（DSP）生成用于细胞内DNA释放的交联聚（乙烯亚胺）（PEI）聚合复合物。二硫键的交联复合物是敏感的细胞内的氧化还原条件和DNA的释放，观察使用溴化乙锭排斥试验和动态光散射。进行转染实验以阐明细胞外和细胞内氧化还原条件的影响。比较了未交联和交联的聚合物的药代动力学和器官蓄积，并在静脉注射后24 h在小鼠中测量基因表达模式。交联的PEI和质粒DNA形成稳定的聚合物，其尺寸范围为100-300 nm，zeta电位在+16 A和+26.1 mV之间。DNA释放发生在二硫键裂解后。还原条件下的细胞培养实验以及谷胱甘肽负载细胞证实了所提出的细胞内活化。细胞内谷胱甘肽状态对转染效率的显着影响observed. Pharmacodynamicprofile的交联PEI/DNA polyplexes在小鼠静脉给药后显示出较高的血液中的交联polyplexes的水平。这些复合物主要积聚在肝和肺中。体内转染数据显示肺转染显著减少（不需要的），而肝转染占主导地位。这些研究表明，交联的聚合复合物在循环中更稳定，并且在静脉内施用后保持其转染效率。(c)2007 Elsevier B. V.保留所有权利。
Crosslinked poly(ethylene imine) (PEI) polyplexes for intracellular DNA release were generated using a low molecular weight crosslinking reagent, Dithiobis(succinimidyl propionate) (DSP). Disulfide bonds of the crosslinked polyplexes were susceptible to intracellular redox conditions and DNA release was observed using an ethidium bromide exclusion assay and dynamic light scattering. Transfection experiments were performed to elucidate the effect of extra- and intracellular redox conditions. Pharmacokinetics and organ accumulation of uncrosslinked and crosslinked polyplexes were compared and gene expression patterns were measured in mice 24 h after intravenous injection.Crosslinked PEI and plasmid DNA formed stable polyplexes in a size range of 100-300 nm, with zeta potentials between + 16A and +26.1 mV. DNA release occurred after cleavage of the disulfide bonds. Cell culture experiments under reducing conditions as well as with glutathione loaded cells confirmed the proposed intracellular activation. A significant influence of the intracellular glutathione status on the transfection efficiency was observed.Pharmacokinetic profiles of crosslinked PEI/DNA polyplexes in mice after intravenous administration showed higher blood levels for crosslinked polyplexes. These polyplexes accumulated mainly in the liver and the lungs. In vivo transfection data revealed significantly reduced (unwanted) lung transfection while liver transfection predominated. These studies suggest that crosslinked polyplexes are more stable in circulation and retain their transfection efficiency after intravenous administration. (c) 2007 Elsevier B.V. All rights reserved.