Multifunctional magnetic nanocarriers for delivery of siRNA and shRNA plasmid to mammalian cells: Characterization, adsorption and release behaviors

Chi-Hsien Liu; Cheng-Han Lin; Yi-Jun Chen; Wei-Chi Wu; Chun-Chao Wang

doi:10.1016/j.colsurfb.2022.112861

Multifunctional magnetic nanocarriers for delivery of siRNA and shRNA plasmid to mammalian cells: Characterization, adsorption and release behaviors

Colloids Surf B Biointerfaces. 2022 Nov:219:112861. doi: 10.1016/j.colsurfb.2022.112861. Epub 2022 Sep 20.

Authors

Chi-Hsien Liu¹, Cheng-Han Lin², Yi-Jun Chen², Wei-Chi Wu³, Chun-Chao Wang⁴

Affiliations

¹ Department of Chemical and Materials Engineering, Chang Gung University, 259, Wen-Hwa First Road, Kwei-Shan, Taoyuan 33302, Taiwan; Research Center for Chinese Herbal Medicine and Research Center for Food and Cosmetic Safety, College of Human Ecology, Chang Gung University of Science and Technology, 261, Wen-Hwa First Road, Taoyuan 33302, Taiwan; Department of Chemical Engineering, Ming Chi University of Technology, 84, Gung-Juan Road, New Taipei City 24301, Taiwan; Department of Ophthalmology, Chang Gung Memorial Hospital, Linkou, 5, Fu-Hsing Street, Taoyuan 33305, Taiwan. Electronic address: chl@mail.cgu.edu.tw.
² Department of Chemical and Materials Engineering, Chang Gung University, 259, Wen-Hwa First Road, Kwei-Shan, Taoyuan 33302, Taiwan.
³ Department of Ophthalmology, Chang Gung Memorial Hospital, Linkou, 5, Fu-Hsing Street, Taoyuan 33305, Taiwan; College of Medicine, Chang Gung University, Taoyuan 33302, Taiwan. Electronic address: weichi666@gmail.com.
⁴ Institute of Molecular Medicine & Department of Medical Science, National Tsing Hua University, 101, Kuang-Fu Road, Hsinchu 30013, Taiwan. Electronic address: ccwang@life.nthu.edu.tw.

PMID: 36162177
DOI: 10.1016/j.colsurfb.2022.112861

Abstract

Nucleic acids are promising candidates for treating various diseases. Nucleic acid is negatively charged and hydrophilic; therefore, it is not efficiently taken up by cells. Successful gene therapy requires the development of carriers for efficient delivery of gene-expressing DNA plasmid and small interfering RNA (siRNA) duplex. In this study, we developed MNP-CA-PEI, a citric acid (CA)-modified magnetic nanoparticle (MNP) cross-linked with polyethyleneimine (PEI), using carbonyldiimidazole as the crosslinker. The physical properties of MNP-CA-PEI (particle size, morphologies, surface coating, surface potentials, magnetic hystereses, superparamagnetic behaviors, and infrared spectra) were systematically characterized by transmission electron microscopy imaging, dynamic light scattering, thermogravimetric analysis, superconducting quantum interference device, and Fourier transform infrared spectroscopy. The adsorption isotherm and kinetics were determined by the Langmuir model, the Freundlich model, a pseudo-first-order equation, and a pseudo-second-order equation. MNP-CA-PEI could form polyelectrolyte complexes with negatively charged nucleic acids, enabling the efficient delivery of nucleic acids into cells. Using MNP-CA-PEI nanoparticles, we magnetically triggered the intracellular delivery of green fluorescence protein (GFP)-expressing DNA plasmid, plasmid-expressing short hairpin RNA (shRNA) against GFP, or siRNA targeting GFP into different cell lines. Nucleic acid/MNP-CA-PEI displayed the enhanced cellular uptake of GFP-expressing DNA plasmid, and it improved the silencing efficiency of shRNA and siRNA, determined by fluorescence imaging. Gene knockdowns mediated by shRNA and siRNA were also confirmed by a quantitative real-time polymerase chain reaction. MNP-CA-PEI delivered nucleic acids into cytosol through caveolae-mediated endocytosis. This study introduces a new MNP functionalization that can be used for the magnetically driven intracellular delivery of nucleic acids.

Keywords: Delivery; Magnetofection; Polyethyleneimine; ShRNA; SiRNA.