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

doi:10.1016/j.jconrel.2009.08.019

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^*

^*此作品的通信作者

生物藥學研究所

研究成果: Article › 同行評審

76 引文斯高帕斯（Scopus）

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 G3139 (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 (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.

原文	English
頁（從 - 到）	62-69
頁數	8
期刊	Journal of Controlled Release
卷	141
發行號	1
DOIs	https://doi.org/10.1016/j.jconrel.2009.08.019
出版狀態	Published - 4 1月 2010

存取文件

10.1016/j.jconrel.2009.08.019

其它文件與鏈接

Link to publication in Scopus

引用此

Koh, C. G., Zhang, X., Liu, S., Golan, S., Yu, B., Yang, X., Guan, J., Jin, Y., Talmon, Y., Muthusamy, N., Chan, K. K., Byrd, J. C., Lee, R. J., Marcucci, G., & Lee, L. J. (2010). Delivery of antisense oligodeoxyribonucleotide lipopolyplex nanoparticles assembled by microfluidic hydrodynamic focusing. Journal of Controlled Release, 141(1), 62-69. https://doi.org/10.1016/j.jconrel.2009.08.019

@article{8a46babc16aa4652a08b7929d187461d,

title = "Delivery of antisense oligodeoxyribonucleotide lipopolyplex nanoparticles assembled by microfluidic hydrodynamic focusing",

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.",

keywords = "Antisense ODN, Bcl-2 down-regulation, G3139, Lipopolyplex, Liposome, Microfluidics, Nanoparticle",

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

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 ). ",

year = "2010",

month = jan,

day = "4",

doi = "10.1016/j.jconrel.2009.08.019",

language = "English",

volume = "141",

pages = "62--69",

journal = "Journal of Controlled Release",

issn = "0168-3659",

publisher = "Elsevier",

number = "1",

}

Koh, CG, Zhang, X, Liu, S, Golan, S, Yu, B, Yang, X, Guan, J, Jin, Y, Talmon, Y, Muthusamy, N, Chan, KK, Byrd, JC, Lee, RJ, Marcucci, G & Lee, LJ 2010, 'Delivery of antisense oligodeoxyribonucleotide lipopolyplex nanoparticles assembled by microfluidic hydrodynamic focusing', Journal of Controlled Release, 卷 141, 編號 1, 頁 62-69. https://doi.org/10.1016/j.jconrel.2009.08.019

TY - JOUR

T1 - Delivery of antisense oligodeoxyribonucleotide lipopolyplex nanoparticles assembled by microfluidic hydrodynamic focusing

AU - Koh, Chee Guan

AU - Zhang, Xulang

AU - Liu, Shujun

AU - Golan, Sharon

AU - Yu, Bo

AU - Yang, Xiaojuan

AU - Guan, Jingjiao

AU - Jin, Yan

AU - Talmon, Yeshayahu

AU - Muthusamy, Natarajan

AU - Chan, Kenneth K.

AU - Byrd, John C.

AU - Lee, Robert J.

AU - Marcucci, Guido

AU - Lee, L. James

N1 - 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 ).

PY - 2010/1/4

Y1 - 2010/1/4

N2 - 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.

AB - 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.

KW - Antisense ODN

KW - Bcl-2 down-regulation

KW - G3139

KW - Lipopolyplex

KW - Liposome

KW - Microfluidics

KW - Nanoparticle

UR - http://www.scopus.com/inward/record.url?scp=70849083668&partnerID=8YFLogxK

U2 - 10.1016/j.jconrel.2009.08.019

DO - 10.1016/j.jconrel.2009.08.019

M3 - Article

C2 - 19716852

AN - SCOPUS:70849083668

SN - 0168-3659

VL - 141

SP - 62

EP - 69

JO - Journal of Controlled Release

JF - Journal of Controlled Release

IS - 1

ER -

Delivery of antisense oligodeoxyribonucleotide lipopolyplex nanoparticles assembled by microfluidic hydrodynamic focusing

摘要

存取文件

其它文件與鏈接

指紋

引用此