Delivery of antisense oligodeoxyribonucleotide lipopolyplex nanoparticles assembled by microfluidic hydrodynamic focusing

JOURNAL OF CONTROLLED RELEASE

卷:141

Article

Koh, Chee Guan¹  Zhang, Xulang²  Liu, Shujun^3,4  Golan, Sharon⁵  Yu, Bo^1,2  Yang, Xiaojuan⁶  Guan, Jingjiao²  Jin, Yan²  Talmon, Yeshayahu⁵  Muthusamy, Natarajan^3,4  Chan, Kenneth K.^3,6  Byrd, John C.^3,4  Lee, Robert J.^2,3,6  Marcucci, Guido^3,4  Lee, L. James^1,2 

[1] Ohio State Univ, Dept Chem & Biomol Engn, Columbus, OH 43210 USA

[2] Ohio State Univ, NSF Nanoscale Sci & Engn Ctr NSEC CANPD, Columbus, OH 43210 USA

[3] Ohio State Univ, Ctr Comprehens Canc, Columbus, OH 43210 USA

[4] Ohio State Univ, Dept Internal Med, Dept Hematol & Oncol, Columbus, OH 43210 USA

[5] Technion Israel Inst Technol, Dept Chem Engn, IL-32000 Haifa, Israel

[6] Ohio State Univ, Coll Pharm, Div Pharmaceut, Columbus, OH 43210 USA

关键词: Microfluidics;    Liposome;    Antisense ODN;    G3139;    Lipopolyplex;    Bcl-2 down-regulation;    Nanoparticle;   

DOI  :  10.1016/j.jconrel.2009.08.019

来源: Elsevier

【 摘 要 】

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.

【 授权许可】

Free