In this work, we examined the effects of the surface charge of stem cell membranes and DNA/polyethyleneimine (PEI) nanocomplexes on gene transfection efficiency, because PEI was one of the most reliable and efficient carriers, and rat bone marrow mesenchymal stem cells (rBMSCs) and rat muscle-derived stem cells (rMDSCs) were one of the readily accessible and plentiful sources of stem cells. Thus, we compared the efficiency of DNA transfection in rBMSCs and rMDSCs using the PEI as a gene carrier. Transfection efficiency was evaluated on the basis of electrostatic interaction between negatively charged stem cell membranes and positively charged DNA/PEI nanocomplexes. DNA was fully complexed with PEI at negative-to-positive (NIP) charge ratios greater than 2, as confirmed by gel electrophoresis and fluorescence measurements. DNA and PEI formed spherical nanocomplexes ranging in diameter from 150 nm to 500 nm. The positive surface charge of DNA/PEI nanocomplexes increased with an increasing N/P charge ratio, as measured using dynamic light scattering and a single-walled carbon nanotube-based field-effect transistor device. rBMSCs and rMDSCs both carried a negative surface charge, with rBMSCs being more negatively charged. The transfection efficiency of rMDSCs measured using DNA/PEI nanocomplexes was very low (1%-5%) at most of the N/P charge ratios tested, whereas better efficiencies were observed with rBMSCs (1%-17%). Nanocomplexes with high NIP charge ratios were cytotoxic to both rBMSCs and rMDSCs. Collectively, the results indicate that rBMSCs were more effectively transfected with DNA/PEI nanocomplexes than were rMDSCs, reflecting the higher negative charge of rBMSC membranes that facilitate the interaction with positively charged DNA/PEI nanocomplexes.
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http://dx.doi.org/10.1166/jbn.2015.2060 | DOI Listing |
Acta Biomater
November 2018
Center for Functional Biomaterials, School of Materials Science and Engineering, and Key Laboratory for Polymeric Composite and Functional Materials of Ministry of Education, Sun Yat-sen University, Guangzhou 510275, China; Department of Materials Science and Engineering, Johns Hopkins University, Baltimore, MD 21218, USA; Institute for NanoBioTechnology, Johns Hopkins University, Baltimore, MD 21218, USA; Department of Biomedical Engineering, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA; Translational Tissue Engineering Center, Johns Hopkins University School of Medicine, Baltimore, MD 21287, USA. Electronic address:
Lipid-based nanoparticles (LNPs) have been developed to address the transport and uptake barriers to enhance the delivery efficiency of plasmid DNA therapeutics. In these systems, plasmid DNA can be encapsulated through condensation by a cationic lipid to form lipo-complexes, or polycation following complexation into cationic liposomes to form lipo-polyplexes. Conventional methods for achieving these two DNA-delivering LNP vehicles suffer from significant batch-to-batch variation, poor scalability and complicated multi-step preparation procedures.
View Article and Find Full Text PDFIn this work, we examined the effects of the surface charge of stem cell membranes and DNA/polyethyleneimine (PEI) nanocomplexes on gene transfection efficiency, because PEI was one of the most reliable and efficient carriers, and rat bone marrow mesenchymal stem cells (rBMSCs) and rat muscle-derived stem cells (rMDSCs) were one of the readily accessible and plentiful sources of stem cells. Thus, we compared the efficiency of DNA transfection in rBMSCs and rMDSCs using the PEI as a gene carrier. Transfection efficiency was evaluated on the basis of electrostatic interaction between negatively charged stem cell membranes and positively charged DNA/PEI nanocomplexes.
View Article and Find Full Text PDFJ Mater Chem B
January 2015
State Key Laboratory of Applied Organic Chemistry, Key Laboratory of Nonferrous Metals Chemistry and Resources Utilization of Gansu Province, College of Chemistry and Chemical Engineering, Lanzhou University, Lanzhou, China.
Polycationic vectors are often used to deliver DNA for cancer therapies, but their inefficiency in releasing DNA from the polyplexes after endosomal escape limits DNA transcription and their efficient application in vivo. In this study, DNA/PEI polyplexes were cross-linked by a reduction-sensitive disulfide bond and then further complexed with electrostatic competitive heparin (HP) and hyaluronidase (HAase)-sensitive hyaluronate (HA) to obtain DNA/PEIS/HA-HP (DPSHA-HP). DPSHA-HP was stable in an extracellular environment (pH = 7.
View Article and Find Full Text PDFACS Appl Mater Interfaces
August 2014
State Key Laboratory of Applied Organic Chemistry, Key Laboratory of Nonferrous Metals Chemistry and Resources Utilization of Gansu Province, College of Chemistry and Chemical Engineering, Institute of Biochemical Engineering & Environmental Technology, Lanzhou University, Lanzhou 730000, China.
Many synthetic Au-based cationic nanoparticles (AuNPs) for nonviral gene delivery show high efficiency in vitro, but their excessive charge density, harsh reducing conditions, and nontarget delivery prevent their application in vivo. Herein, we constructed a sandwich-type layered polyethylenimine (PEI)-coated gold nanocomposite outerlaid with a nucleus-targeted Dexamethasone (Dexa), namely, Au-PEI/DNA/PEI-Dexa nanocomplex, for DNA delivery system using a low molecular weight PEI as a mild reducing agent. The nucleus-targeting Au-PEI/DNA/PEI-Dexa nanocomplex with low positive charge and low cytotoxicity condensed DNA and protected from enzymatic degradation.
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