scholarly journals Synthesis, and Characterization, and Evaluation of Cellular Effects of the FOL-PEG-g-PEI-GAL Nanoparticles as a Potential Non-Viral Vector for Gene Delivery

2010 ◽  
Vol 2010 ◽  
pp. 1-10 ◽  
Author(s):  
S. Ghiamkazemi ◽  
A. Amanzadeh ◽  
R. Dinarvand ◽  
M. Rafiee-Tehrani ◽  
M. Amini
Keyword(s):  
Gene Delivery ◽  
Zeta Potential ◽  
Surface Charge ◽  
Cell Line ◽  
Graft Copolymer ◽  
Viral Vector ◽  
Transfection Efficiency ◽  
High Surface ◽  
Liver Cells ◽  
Dna Complexes

In this manuscript, we synthesized the potential non viral vector for gene delivery with proper transfection efficiency and low cytotoxicity. Polyethylenimine (PEI) is a well-known cationic polymer which has high positive surface charge for condensing plasmid DNA. However; it is highly cytotoxic in many cell lines because of the high surface charge, non-biodegradability and non-biocompatibility. To enhance PEI biodegradability, the graft copolymer “PEG-g-PEI” was synthesized. To target cancer liver cells, two targeting ligands folic acid and galactose (lactobionic acid) which are over expressed on human hepatocyte carcinoma were attached to graft copolymer and “FOL-PEG-g-PEI-GAL” copolymer was synthesized. Composition of this grafted copolymer was characterized using1H-NMR and FTIR spectra. The molecular weight and zeta potential of this copolymer was compared to PEI. The particle size and zeta potential of FOL-PEG-g-PEI-GAL/DNA complexes at various N/P ratio were measured using dynamic light scattering (DLS). Cytotoxicity of the copolymer was also studied in cultured HepG2 human hepatoblastoma cell line. The FOL-PEG-g-PEI-GAL/DNA complexes at various N/P ratios exhibited no cytotoxicity in HepG2 cell line compared to PEI 25K as a control. The novel copolymer showed enhanced biodegradability in physiological conditions in compared with PEI and targeted cultured HepG2 cells. More importantly, significant transfection efficiency was exhibited in cancer liver cells. Together, our results showed that “FOL-PEG-g-PEI-GAL” nanoparticals could be considered as a useful non-viral vector for targeted gene delivery.

Cholic Acid-Modified Polyethylenimine for Gene Delivery into Human Carcinoma Cells

2014 ◽  
Vol 1060 ◽  
pp. 3-6 ◽  
Author(s):  
Wanlop Weecharangsan ◽  
Orapan Paecharoenchai ◽  
Nattisa Niyomtham ◽  
Praneet Opanasopit ◽  
Boon-ek Yingyongnarongkul ◽  
...  
Keyword(s):  
Gene Delivery ◽  
Zeta Potential ◽  
Cholic Acid ◽  
Plasmid Dna ◽  
Gel Electrophoresis ◽  
Transfection Efficiency ◽  
Carcinoma Cells ◽  
Molar Ratio ◽  
Dna Complexes ◽  
Human Carcinoma

Polyethylenimine (PEI) was modified by cholic acid at a molar ratio of 1:1. Cholic acid (CA)-modified PEI (PEI-CA) were evaluated for formation of DNA complexes. PEI-CA/pEGFP plasmid DNA complexes were characterized for their size and zeta potential. Gel electrophoresis showed total retardation for PEI-CA/pEGFP complexes formed at weight ratios above 0.25. The particle size and zeta potential of the complexes at a polymer-to-DNA ratio of 0.5 were 295.3 nm and 30.5 mV, respectively. The transfection efficiency of PEI-CA/pEGFP complexes was comparable to unmodified PEI. Cytotoxicity result showed that PEI-CA had lower cytoxicity than PEI. This study suggests that PEI-CA has potential utility as a gene delivery carrier.

The Influence of Physicochemical Parameters on the Efficacy of Non-Viral DNA Transfection Complexes: A Comparative Study

2006 ◽  
Vol 6 (9) ◽  
pp. 2776-2782 ◽  
Author(s):  
Carsten Kneuer ◽  
Carsten Ehrhardt ◽  
Heike Bakowsky ◽  
M. N. V. Ravi Kumar ◽  
Volker Oberle ◽  
...  
Keyword(s):  
Zeta Potential ◽  
Mammalian Cells ◽  
Transfection Efficiency ◽  
Dna Delivery ◽  
Dna Transfection ◽  
Dna Complexes ◽  
General Validity ◽  
Delivery Vehicles ◽  
Foreign Dna ◽  
Pronounced Reduction

Various polycationic vehicles have been developed to facilitate the transfer of foreign DNA into mammalian cells. Structure-activity studies suggested that biophysical properties, such as size, charge, and morphology of the resulting DNA complexes determine transfection efficiency within one class of vector. To investigate the general validity of these criteria, we studied the efficacy of a variety of DNA delivery vehicles including liposomes (DOTAP, SAINT2) with and without helper lipid (DOPE), the polymer polyethyleneimine (PEI), and cationic nanoparticles (Si26H, PLGA/chitosan) in a comparative manner. Sizes of the DNA complexes varied between 100 and 500 nm for PEI polyplexes and DOTAP/DOPE lipoplexes, respectively. The zeta potential was positive for PEI, Si26H, and DOTAP based complexes, while it was neutral for SAINT2-DNA complexes and negative for PLGA/chitosan-DNA complexes. The latter finding was elucidated by AFM, showing a layer of DNA adsorbed onto the nanoparticles. Transfection activity was negligible for PLGA/chitosan nanospheres, moderate for Si26H nanospheres and high for all other complexes, PEI being the most active carrier. The liposomal preparations were of low (DOTAP) or moderate (SAINT2) stability in serum, resulting in a pronounced reduction of gene expression, which was partially restored by the addition of chloroquine. In conclusion, transfection efficiency (i) seems to require a positive or neutral zeta potential, (ii) is depending on size, e.g., is higher for smaller particles, and (iii) requires a vector that is stable in serum.

CATIONIC POLYMER BASED GENE DELIVERY: UPTAKE AND INTRACELLULAR TRAFFICKING

COSMOS ◽  
2014 ◽  
Vol 10 (01) ◽  
pp. 17-24
Author(s):  
YOONKHEI HO ◽  
HENG-PHON TOO
Keyword(s):  
Gene Delivery ◽  
Transfection Efficiency ◽  
Cell Types ◽  
Endosomal Escape ◽  
Current Status ◽  
Uptake Mechanism ◽  
Nuclear Entry ◽  
Dna Complexes ◽  
Major Drawback ◽  
Gene Carriers

To date, low transfection efficiency remains the major drawback of polymer based gene delivery. Many cell types including stem cells, fibroblast and neurons are known to be poorly transfected with polymer based gene carriers and the high toxicity severely restrict their utility in gene delivery. Continual efforts are made to identify cellular barriers to efficient transfection as these carriers have low immunogenicity, ease of manufacturing and scalability. Here, we summarize the current status of understanding on uptake mechanism of polymer-DNA complexes (polyplexes), their endosomal escape, cytosolic transport and nuclear entry of pDNA.

Novel Poly(Ester Amine) Based on Polycaprolactone and Polyethylenimine as a Gene Carrier: Effect of Hydrophobicity on Transfection Efficiency and Cytotoxicity

2007 ◽  
Vol 342-343 ◽  
pp. 453-456 ◽  
Author(s):  
Rohidas B. Arote ◽  
Tae Hee Kim ◽  
You Kyoung Kim ◽  
Dhananjay Jere ◽  
Hu Lin Jiang ◽  
...  
Keyword(s):  
Gene Delivery ◽  
Transfection Efficiency ◽  
Gene Carrier ◽  
Particle Sizes ◽  
Dna Complexes ◽  
Novel Gene ◽  
Carrier Effect ◽  
Biodegradable Poly ◽  
Low Cytotoxicity

Novel, biodegradable poly(ester amine)s (PEAs) were synthesized using hydrophobic polycaprolactone diacrylate (PCLDA) and highly cationic polyethylenimine (PEI). This novel gene carrier can form stable DNA complexes with particle sizes around 200 nm, and showing excellent transfection efficiency and relatively low cytotoxicity compared with PEI 25K. Effect of hydrophobicity on transfection efficiency and cytotoxicity was profound and was relatively important parameter for the success of gene delivery.

The Effect of Spermidine and Spermine on Chitosan-Mediated Gene Delivery

2020 ◽  
Vol 859 ◽  
pp. 113-119
Author(s):  
Auayporn Apirakaramwong ◽  
Songporn Sunthornphan ◽  
Nutcha Pasuthawong ◽  
Tanawin Intaravicha ◽  
Nattapon Ruangthai ◽  
...  
Keyword(s):  
Gene Delivery ◽  
Viral Vector ◽  
Transfection Efficiency ◽  
Weight Ratio ◽  
Green Fluorescence Protein ◽  
Ternary Complexes ◽  
Average Cell ◽  
Living Organisms ◽  
Almost All

Chitosan (CS) is a biodegradable and highly positive charged polymer that can be utilized as a non-viral vector for gene delivery. However, its transfection efficiency is low. Spermidine (SPD) and spermine (SPM) are ordinary polyamines found in almost all living organisms. Their structures contain tri- and tetra-amine, respectively. The present study aims to elucidate the effect of SPD or SPM on transfection efficiency and cytotoxicity of CS/DNA complexes in human cervical carcinoma (HeLa) cells. The ternary complexes (CS, DNA and SPD or SPM) at different weight ratios and mixing orders were formulated and tested on transfection efficiency. Their particle sizes and charges were also estimated. CS that has a molecular weight of 45,000 Daltons and degree of deacetylation of 85 was able to form complete complex with plasmid DNA expressing enhanced green fluorescence protein (pEGFP-C2) at weight ratio of 4. The SPM/CS/DNA (1:4:1) and SPD/ CS/DNA (512:4:1) complexes illustrated the greatest transfection efficiency. Moreover, the transfection efficiency was affected by the mixing order between CS, DNA and SPD or SPM. For cell toxicity experiments, more than 85% the average cell survival of the complexes were detected by 3-(4,5-dimethylthiazolyl-2)-2,5-diphenyltetrazolium bromide (MTT) cell growth assay. These findings suggest that addition of either SPD or SPM to CS before forming ternary complexes with DNA may significantly improve gene delivery potential in vitro.

Cationic liposome–DNA complexes: from liquid crystal science to gene delivery applications

2006 ◽  
Vol 364 (1847) ◽  
pp. 2573-2596 ◽  
Author(s):  
Cyrus R Safinya ◽  
Kai Ewert ◽  
Ayesha Ahmad ◽  
Heather M Evans ◽  
Uri Raviv ◽  
...  
Keyword(s):  
Gene Delivery ◽  
Laser Scanning ◽  
Liquid Crystalline ◽  
Transfection Efficiency ◽  
Three Dimensional ◽  
Cationic Liposome ◽  
Laser Scanning Confocal Microscopy ◽  
Dna Complexes ◽  
Scanning Confocal Microscopy ◽  
Crystalline Nature

At present, there is an unprecedented level of interest in the properties and structures of complexes consisting of DNA mixed with oppositely charged cationic liposomes (CLs). The interest arises because the complexes mimic natural viruses as chemical carriers of DNA into cells in worldwide human gene therapy clinical trials. However, since our understanding of the mechanisms of action of CL–DNA complexes interacting with cells remains poor, significant additional insights and discoveries will be required before the development of efficient chemical carriers suitable for long-term therapeutic applications. Recent studies describe synchrotron X-ray diffraction, which has revealed the liquid crystalline nature of CL–DNA complexes, and three-dimensional laser-scanning confocal microscopy, which reveals CL–DNA pathways and interactions with cells. The importance of the liquid crystalline structures in biological function is revealed in the application of these modern techniques in combination with functional transfection efficiency measurements, which shows that the mechanism of gene release from complexes in the cell cytoplasm is dependent on their precise liquid crystalline nature and the physical and chemical parameters (for example, the membrane charge density) of the complexes. In §5 , we describe some recent new results aimed at developing bionanotube vectors for gene delivery.

Hepatocyte-Specific Gene Delivery with Galactose-Bearing Cationic Polymers with Different Molecular Structures

2012 ◽  
Vol 86 ◽  
pp. 86-91 ◽  
Author(s):  
Maria Chiara Munisso ◽  
Atsushi Mahara ◽  
Yoichi Tachibana ◽  
Jeong Hun Kang ◽  
Satoshi Obika ◽  
...  
Keyword(s):  
Gene Delivery ◽  
Viral Vector ◽  
Transfection Efficiency ◽  
Molecular Structures ◽  
Specific Gene ◽  
Cationic Polymers ◽  
Gene Therapies ◽  
Polymeric Carriers ◽  
Significant Efficacy

Since the promising virus -based gene therapies are often limited by problems such as the immunity of virus itself, the development of an efficient non-viral vector is of prime importance. For this reason, several synthetic nonviral polymeric carriers including cationic sequences have been molecularly designed. It is well known that the polymeric carriers with some cationic groups buffer the endosomal pH resulting in the enhanced transfection efficiency, but also in a relatively high toxicity. In the last decades, the polymers bearing pendant carbohydrates (glycopolymers) was proved to have relatively less toxic. Since the glycopolymers may not only decrease the toxicity of the cationic chain but also serve as targeting agent, we have rationally designed new glycopolymer-based gene delivery carriers. The interaction of carrier/gene polyplexes with hepatocytes and their intracellular trafficking were investigated in vitro. Our results show the significant efficacy of the galactose moieties on the uptake by hepatocytes, in a ligand specific manner.

scholarly journals pH-Sensitive Pegylated Cationic Lipid-DNA Complexes for Gene Delivery: Transfection Efficiency and Live Cell Imaging Studies

2012 ◽  
Vol 102 (3) ◽  
pp. 501a-502a ◽  
Author(s):  
Chia-Ling Chan ◽  
Ramsey Majzoub ◽  
Rahau S. Shirazi ◽  
Keng-San Liang ◽  
Kai K. Ewert ◽  
...  
Keyword(s):  
Gene Delivery ◽  
Live Cell Imaging ◽  
Cell Imaging ◽  
Transfection Efficiency ◽  
Cationic Lipid ◽  
Ph Sensitive ◽  
Live Cell ◽  
Imaging Studies ◽  
Dna Complexes

Cationic Gelatin as a Gene Carrier

1995 ◽  
Vol 394 ◽  
Author(s):  
K. E. Brown ◽  
J. Bathon ◽  
C. H. Huang ◽  
R. Dalai ◽  
K. W. Leong
Keyword(s):  
Mammalian Cells ◽  
Viral Vector ◽  
Transfection Efficiency ◽  
Cell Types ◽  
Cell Transfection ◽  
Serum Free Medium ◽  
Free Medium ◽  
Mobility Shift ◽  
Dna Complexes ◽  
Dye Reduction

AbstractCationic gelatin was evaluated as a non-viral vector for cell transfection. We hypothesized that cationic gelatin would be a nontoxic alternative to already existing viral and non-viral cationic vectors. Cationic gelatin was synthesized by modifying gelatin with hexanediamine. Complexation of cationic gelatin with psv-β-gal plasmid caused an electrophoretic mobility shift of the plasmid. Cationic gelatin/DNA complexes were optimized in terms of transfection efficiency in CHODUK XB1 and COS 7 cell lines. Maximal gene expression for both cell types occurred in serum free medium with chloroquine (100 μM) at cationic gelatin/DNA ratios of approximately 2 and 7. In comparison with DEAE dextran, polylysine and Lipofectamine, cationic gelatin was the most efficient in transfecting COS 7 cells, with up to 18% cells transfected. In a dye reduction cytotoxicity assay, cationic gelatin caused < 5% of cells to become nonviable at a concentration of 100 μg/ml, while the other transfection reagents tested at the same concentration caused 25–100% of cell death. These results suggest that cationic gelatin holds promise as an effective vehicle for gene delivery to mammalian cells.

Glucosylated Polypropylenimine Dendrimer as a Novel Gene Carrier

2007 ◽  
Vol 342-343 ◽  
pp. 457-460 ◽  
Author(s):  
You Kyoung Kim ◽  
In Kyu Park ◽  
Hu Lin Jiang ◽  
Rohidas B. Arote ◽  
Hwan Jeong Jeong ◽  
...  
Keyword(s):  
Sulfuric Acid ◽  
Dna Binding ◽  
Gene Delivery ◽  
Hela Cell ◽  
Cell Line ◽  
Plasmid Dna ◽  
Transfection Efficiency ◽  
Hela Cell Line ◽  
Binding Ability ◽  
Low Cytotoxicity

Polypropylenimine (PPI) dendrimers have been used by many researchers as gene delivery carriers due to their high functionality. Glucose as a kind of carbohydrate is biocompatible and hydrophilic. In this study, we synthesized glucosylated PPI (G-PPI) dendrimers to reduce cytotoxicity. Glucose substitution of G-PPI dendrimers was determined by the sulfuric acid micromethod. The G-PPI dendrimer was complexed with plasmid DNA in various N/P ratios, and the complex was characterized. G-PPI dendrimers showed good DNA binding ability and high protection of DNA from nuclease attack. The G-PPI dendrimer had low cytotoxicity compared to PPI dendrimer by cytotoxicity assay. Also, transfection efficiency was influenced by glucosylation degree and the transfection efficiency for the G-PPI-5 was slightly higher than that of PPI dendrimer in HeLa cell line.

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