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Fabrication of a Novel Core-Shell Gene Delivery System Based on a Brush-Like Polycation of α, β–Poly (L-Aspartate-Graft-PEI)

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Abstract

Purpose

A novel core-shell gene delivery system was fabricated in order to improve its gene transfection efficiency, particularly in the presence of serum.

Materials and Methods

α, β–poly (L-aspartate-graft-PEI) (PAE) was simply synthesized by ring-opening reaction of poly (L-succinimide) with low molecular weight (LMW) linear polyethylenimine (PEI, Mn = 423). PAE/DNA nanoparticles were characterized. Condensation and protection ability of plasmid by PAE were confirmed by agarose gel electrophoresis assay. Cytotoxicity of the polymer and polymer/DNA nanoparticles were measured by MTS assay. Gene transfection efficiencies were evaluated both in vitro and in vivo.

Results

Core-shell nanoparticles assembled between DNA and PAE showed positive zeta potential, narrow size distribution, and spherical compact shapes with size below 250 nm when N/P ratio is above 10. Cytotoxicity of PAE was rather lower than that of PEI 25K, while the most efficient gene transfection and serum resistant ability of PAE/DNA complexes were higher than that of PEI 25K. Bafilomycin A1 treatment suggested “proton sponge” mechanism of PAE-mediated gene transfection. PAE/pEGFP-N2 nanoparticles also showed good gene expression in vivo and were dominantly distributed in kidney, liver, spleen and lung after intravenous administration.

Conclusions

The results demonstrated the potential use of PAE as an effective gene carrier.

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Abbreviations

DCC:

N, N′-dicyclohexylcarbodiimide

DMEM:

Dulbecco’s modified Eagle’s medium

DMF:

N, N-dimethylformamide

DMSO:

dimethyl sulfoxide

EGFP:

enhanced green fluorescent protein

FBS:

fetal bovine serum

HGF:

(hepatocyte growth factor)

LMW:

low molecular weight

MPS:

mononuclear phagocytic system

PAE:

α, β–poly (L-aspartate-graft-PEI)

PEI:

polyethylenimine

PSI:

poly (L-succinimide)

RLUs:

Relative light units

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Acknowledgements

This work was financially supported by Korea Science and Engineering Foundation (ROI-2005-000-10087-0) and Natural Science Foundation of China (No. 20504010). We also acknowledge the National Instrumentation Center for Environmental Management (NICEM) for permission to take NMR measurements. J. H. Yu was supported by KOSEF as a post-doctor under Korea-China Young Scientist Exchange Program, and J.S. Quan was supported by BK21 program.

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Authors and Affiliations

  1. Department of Agricultural Biotechnology, Seoul National University, Seoul, 151-921, South Korea

    Jia-Hui Yu, Ji-Shan Quan & Chong-Su Cho

  2. College of New Drug Innovation Research & Development, East China Normal University, Shanghai, 200062, P. R. China

    Jia-Hui Yu

  3. College of Veterinary Medicine, Seoul National University, Seoul, 151-742, South Korea

    Jung-Taek Kwon, Cheng-Xiong Xu, Bo Sun, Hu-Lin Jiang & Myung-Haing Cho

  4. College of Pharmacy, Yanbian University, Jilin Province, 133000, P. R. China

    Ji-Shan Quan

  5. Research Institute of Agriculture and Life Sciences, Seoul National University, Seoul, 151-921, South Korea

    Chong-Su Cho

  6. Department of Polymer Science and Engineering, Sunchon National University, Sunchon, 540-742, South Korea

    Jae-Woon Nah

  7. Department of Nuclear Medicine, Research Institute of Clinical Medicine, Chonbuk National University School of Medicine, Jeonju, 561-712, South Korea

    Eun-Mi Kim & Hwan-Jeong Jeong

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  1. Jia-Hui Yu
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Corresponding authors

Correspondence to Myung-Haing Cho or Chong-Su Cho.

Additional information

J.-H. Yu and J.-S. Quan have contributed equally to this work.

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Yu, JH., Quan, JS., Kwon, JT. et al. Fabrication of a Novel Core-Shell Gene Delivery System Based on a Brush-Like Polycation of α, β–Poly (L-Aspartate-Graft-PEI). Pharm Res 26, 2152–2163 (2009). https://doi.org/10.1007/s11095-009-9928-9

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  • DOI: https://doi.org/10.1007/s11095-009-9928-9

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