Delivery of antisense oligodeoxyribonucleotide lipopolyplex nanoparticles assembled by microfluidic hydrodynamic focusing

Chee Guan Koh, Xulang Zhang, Shujun Liu, Sharon Golan, Bo Yu, Xiaojuan Yang, Jingjiao Guan, Yan Jin, Yeshayahu Talmon, Natarajan Muthusamy, Kenneth K. Chan, John C. Byrd, Robert J. Lee, Guido Marcucci, L. James Lee

Research output: Contribution to journalArticlepeer-review

51 Scopus citations

Abstract

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 GenasenseTM) 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.

Original languageEnglish (US)
Pages (from-to)62-69
Number of pages8
JournalJournal of Controlled Release
Volume141
Issue number1
DOIs
StatePublished - Jan 4 2010
Externally publishedYes

Bibliographical note

Funding Information:
We thank Le-Nguyen Huynh and Jiuxia Pang for helpful discussion on real-time RT-PCR and Western blots. We are grateful to NSF NSEC-CANPBD (Award No. EEC-0425626 ) and NIH for financial support ( R01: CA135243 and R21: 131832 ).

Keywords

  • Antisense ODN
  • Bcl-2 down-regulation
  • G3139
  • Lipopolyplex
  • Liposome
  • Microfluidics
  • Nanoparticle

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