Cationic lipid-nanoceria hybrids, a novel nonviral vector-mediated gene delivery into mammalian cells: investigation of the cellular uptake mechanism.

Joydeep Das,Hyuk Song,Chankyu Park,Han Geuk Seo,Ssang-Goo Cho,Jin-Hoi Kim,Jae Woong Han,Yun-Jung Choi

doi:10.1038/srep29197

Abstract

Gene therapy is a promising technique for the treatment of various diseases. The development of minimally toxic and highly efficient non-viral gene delivery vectors is the most challenging undertaking in the field of gene therapy. Here, we developed dimethyldioctadecylammonium bromide (DODAB)–nanoceria (CeO2) hybrids as a new class of non-viral gene delivery vectors. These DODAB-modified CeO2 nanoparticles (CeO2/DODAB) could effectively compact the pDNA, allowing for highly efficient gene transfection into the selected cell lines. The CeO2/DODAB nanovectors were also found to be non-toxic and did not induce ROS formation as well as any stress responsive and pro-survival signaling pathways. The overall vector performance of CeO2/DODAB nanohybrids was comparable with lipofectamine and DOTAP, and higher than calcium phosphate and DEAE-dextran for transfecting small plasmids. The increased cellular uptake of the nanovector/DNA complexes through clathrin- and caveolae-mediated endocytosis and subsequent release from the endosomes further support the increased gene transfection efficiency of the CeO2/DODAB vectors. Besides, CeO2/DODAB nanovectors could transfect genes in vivo without any sign of toxicity. Taken together, this new nano-vector has the potential to be used for gene delivery in biomedical applications.

Highlights

IntroductionWe propose that by combining the advantage of dimethyldioctadecylammonium bromide (DODAB) as a gene delivery vector and the biocompatible nature of nanoceria to prepare a one-particle system (CeO2/DODAB), high gene delivery efficiency can be achieved
The synthesized CeO2 was characterized by energy dispersive spectroscopy (EDS) and Fourier transform infrared (FTIR) spectroscopic analyses
We prepared a new class of nanoceria (CeO2)-based gene delivery vectors, and dimethyldioctadecylammonium bromide (DODAB) was utilized for surface modification of negatively charged CeO2 nanoparticles via simple electrostatic interaction

Summary

Introduction

We propose that by combining the advantage of DODAB as a gene delivery vector and the biocompatible nature of nanoceria to prepare a one-particle system (CeO2/DODAB), high gene delivery efficiency can be achieved To test this hypothesis we (i) prepared negatively charged nanoceria from ammonium cerium(IV) nitrate with subsequent characterization, (ii) characterized positively charged nanoceria prepared via surface modification using DODAB, (iii) evaluated the size and surface zeta potentials of the nanoparticles and nanoparticle-DNA complexes, (iv) assessed the DNA complexing ability and protection against DNase I; (v) checked the in vitro transfection efficiency and cytocompatibility of the nanoparticles and intracellular distribution of the nanoparticle-DNA complexes, (vi) investigated the intracellular uptake pathways of the nanoparticle-DNA complexes, and (vii) evaluated the in vivo transfection efficiency and biocompatibility of the nanoparticles

Methods

Results

Conclusion