Delivery of antisense oligodeoxyribonucleotide lipopolyplex nanoparticles assembled by microfluidic hydrodynamic focusing.
Delivery of antisense oligodeoxyribonucleotide lipopolyplex nanoparticles assembled by microfluidic hydrodynamic focusing.
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DOI：
10.1016/j.jconrel.2009.08.019
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发表时间：
2010-01-04
影响因子：
10.8
通讯作者：
Lee, L. James
中科院分区：
文献类型：
--
作者：
Koh, Chee Guan;Zhang, Xulang;Liu, Shujun;Golan, Sharon;Yu, Bo;Yang, Xiaojuan;Guan, Jingjiao;Jin, Yan;Talmon, Yeshayahu;Muthusamy, Natarajan;Chan, Kenneth K.;Byrd, John C.;Lee, Robert J.;Marcucci, Guido;Lee, L. James
建立了一种多入口微流控流体动力聚焦（MF）系统，用于制备含Bcl-2反义脱氧寡核苷酸（ODN）的脂质复合物（LP）。脂质多聚物纳米颗粒由ODN：鱼精蛋白：脂质（1：0.3：12.5重量/重量比）组成，脂质包括DC-Chol：卵PC：PEG-DSPE（40：58：2 mol/mol%）。使用K562人红白血病细胞（其含有丰富的Bcl-2和转铁蛋白受体（TfR）的过表达）和G3139（oblimerson sodium或Genasense™）分别作为模型细胞系和药物，在常规批量混合（BM）方法和MF方法之间比较了mRNA和蛋白质水平上的Bcl-2下调以及细胞摄取和凋亡。通过动态光散射（DLS）和Zeta电位（Zeta potential，Zeta）测量分别表征脂质多聚物的大小和表面电荷，同时通过凝胶电泳测定ODN的包封率。低温透射电子显微镜（Cryo-TEM）用于确定LP的形态。我们的研究结果表明，MF生产的LP纳米粒子具有相似的结构，但较小的尺寸和尺寸分布相比，BM LP纳米粒子。MF LP纳米粒具有更高水平的Bcl-2反义摄取，并且显示出比BM LP纳米粒更有效的Bcl-2蛋白水平下调。
A multi-inlet microfluidic hydrodynamic focusing (MF) system to prepare lipopolyplex (LP) containing Bcl-2 antisense deoxyoligonucleotide (ODN) was developed and evaluated. The lipopolyplex nanoparticles consist of ODN:protamine:lipids (1:0.3:12.5 wt/wt ratio) and the lipids included DC-Chol:egg PC:PEG–DSPE (40:58:2 mol/mol%). Using K562 human erythroleukemia cells, which contain an abundance of Bcl-2 and overexpression of transferrin receptors (TfR), and G3139 (oblimerson sodium or Genasense™) as a model cell line and drug, respectively, the Bcl-2 down-regulation at the mRNA and protein levels as well as cellular uptake and apoptosis was compared between the conventional bulk mixing (BM) method and the MF method. The lipopolyplex size and surface charge were characterized by dynamic light scattering (DLS) and zeta potential (ζ) measurement, respectively, while the ODN encapsulation efficiency was determined by gel electrophoresis. Cryogenic transmission electron microscopy (Cryo-TEM) was used to determine the morphology of LPs. Our results demonstrated that MF produced LP nanoparticles had similar structures but smaller size and size distribution compared to BM LP nanoparticles. MF LP nanoparticles had higher level of Bcl-2 antisense uptake and showed more efficient down-regulation of Bcl-2 protein level than BM LP nanoparticles.
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影响因子：
15
作者：
Jahn, A;Vreeland, WN;Locascio, LE
通讯作者：
Locascio, LE
影响因子：
4.8
作者：
Sato, Y;Yamauchi, N;Niitsu, Y
通讯作者：
Niitsu, Y
影响因子：
3.9
作者：
Jahn, Andreas;Vreeland, Wyatt N.;Gaitan, Michael
通讯作者：
Gaitan, Michael
影响因子：
4.9
作者：
Yang X;Koh CG;Liu S;Pan X;Santhanam R;Yu B;Peng Y;Pang J;Golan S;Talmon Y;Jin Y;Muthusamy N;Byrd JC;Chan KK;Lee LJ;Marcucci G;Lee RJ
通讯作者：
Lee RJ
影响因子：
--
作者：
Kerkmann, Miren;Lochmann, Dirk;Hartmann, Gunther
通讯作者：
Hartmann, Gunther