期刊
JOURNAL OF CONTROLLED RELEASE
卷 141, 期 1, 页码 62-69出版社
ELSEVIER SCIENCE BV
DOI: 10.1016/j.jconrel.2009.08.019
关键词
Microfluidics; Liposome; Antisense ODN; G3139; Lipopolyplex; Bcl-2 down-regulation; Nanoparticle
资金
- NSF [EEC-0425626]
- NIH [R01: CA735243, R21: 131832]
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 63139 (oblimerson sodium or Genasense (TM)) 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 (zeta) 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. (C) 2009 Elsevier B.V. All rights reserved.
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