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.

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.

scholarly journals Direct visualization of the binding and internalization of a ferritin conjugate of epidermal growth factor in human carcinoma cells A-431.

1979 ◽  
Vol 81 (2) ◽  
pp. 382-395 ◽  
Author(s):  
H T Haigler ◽  
J A McKanna ◽  
S Cohen
Keyword(s):  
Plasma Membrane ◽  
Growth Factor ◽  
Epidermal Growth Factor ◽  
Carcinoma Cells ◽  
Biologically Active ◽  
Molar Ratio ◽  
Multivesicular Bodies ◽  
Egf Receptors ◽  
Human Carcinoma ◽  
Epidermal Growth

We have prepared a conjugate of epidermal growth factor (EGF) and ferritin that retains substantial binding affinity for cell receptors and is biologically active. Glutaraldehyde-activated EGF was covalently linked to ferritin to produce a conjugate that contained EGF and ferritin in a 1:1 molar ratio. The conjugate was separated from free ferritin by affinity chromatography using antibodies to EGF. Monolayers of human epithelioid carcinoma cells (A-431) were incubated with EGF:ferritin at 4 degrees C and processed for transmission electron microscopy. Under these conditions, approximately 6 X 10(5) molecules of EGF:ferritin bound to the plasma membrane of each cell. In the presence of excess native EGF, the number of bound ferritin particles was reduced by 99%, indicating that EGF:ferritin binds specifically to cellular EGF receptors. At 37 degrees C, cell-bound EGF:ferritin rapidly redistributed in the plane of the plasma membrane to form small groups that were subsequently internalized into pinocytic vesicles. By 2.5 min at 37 degrees C, 32% of the cell-bound EGF:ferritin was localized in vesicles. After 2.5 min, there was a decrease in the proportion of conjugate in vesicles with a concomitant accumulation of EGF:ferritin in multivesicular bodies. By 30 min, 84% of the conjugate was located in structures morphologically identified as multivesicular bodies or lysosomes. These results are consistent with other morphological and biochemical studies utilizing 125I-EGF and fluorescein-conjugated EGF.

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.

scholarly journals In Vitro Gene Delivery Mediated by Asialofetuin-Appended Cationic Liposomes Associated with γ-Cyclodextrin into Hepatocytes

2011 ◽  
Vol 2011 ◽  
pp. 1-13 ◽  
Author(s):  
Keiichi Motoyama ◽  
Yoshihiro Nakashima ◽  
Yukihiko Aramaki ◽  
Fumitoshi Hirayama ◽  
Kaneto Uekama ◽  
...  
Keyword(s):  
Gene Transfer ◽  
Gene Delivery ◽  
Plasmid Dna ◽  
Transfection Efficiency ◽  
Cationic Liposomes ◽  
Nonviral Vector ◽  
Liposomal Membrane ◽  
Transfer Activity ◽  
Potential Use

The purpose of this study is to evaluate in vitro gene delivery mediated by asialofetuin-appended cationic liposomes (AF-liposomes) associating cyclodextrins (CyD/AF-liposomes) as a hepatocyte-selective nonviral vector. Of various CyDs, AF-liposomes associated with plasmid DNA (pDNA) and γ-cyclodextrin (γ-CyD) (pDNA/γ-CyD/AF-liposomes) showed the highest gene transfer activity in HepG2 cells without any significant cytotoxicity. In addition, γ-CyD enhanced the encapsulation ratio of pDNA with AF-liposomes, and also increased gene transfer activity as the entrapment ratio of pDNA into AF-liposomes was increased. γ-CyD stabilized the liposomal membrane of AF-liposomes and inhibited the release of calcein from AF-liposomes. The stabilizing effect of γ-CyD may be, at least in part, involved in the enhancing gene transfer activity of pDNA/γ-CyD/AF-liposomes. Therefore, these results suggest the potential use of γ-CyD for an enhancer of transfection efficiency of AF-liposomes.

PEGylated Plier-Like Cationic Niosomes on Gene Delivery in HeLa Cells

2019 ◽  
Vol 819 ◽  
pp. 151-156
Author(s):  
Supusson Pengnam ◽  
Samarwadee Plianwong ◽  
Kanokwan Singpanna ◽  
Nattisa Ni-yomtham ◽  
Widchaya Radchatawedchakoon ◽  
...  
Keyword(s):  
Particle Size ◽  
Gene Delivery ◽  
Hela Cells ◽  
Transfection Efficiency ◽  
Cationic Lipid ◽  
Dna Delivery ◽  
Molar Ratio ◽  
The Novel ◽  
Low Cytotoxicity ◽  
Cationic Niosomes

Lipid-based formulations have been used as a widespread carrier to improve gene delivery. Niosomes, one type of lipid-based vesicular systems are produced from non-ionic surfactants which are generally inexpensive and potentially more stable than phospholipids. This article was to develop PEGylated cationic niosomes for DNA delivery. Thin film hydration and sonication method were applied for cationic niosomes. The niosome formulations were composed of Span 20, cholesterol (Chol) and plier-like cationic lipid B (PCL-B) with or without cholesterol-polyethylene glycol 2000 (Chol-PEG). The physicochemical properties of cationic niosomes and nioplexes were evaluated including particle size, zeta potential, DNA condensation and serum protection. The transfection efficiency and cell viability were examined in HeLa cells. The particle size and surface charge of PEGylated cationic niosome containing Span 20: Chol: PCL-B: Chol-PEG at the molar ratio of 2.5: 2.5: 1.5: 0.14 (N-PEG2) were 129.47 ± 2.15 nm and 25.93 ± 4.18 mV, respectively. These PEGylated cationic niosomes could condense pDNA into the nanosize particles and also enhance the serum protection ability for at least 6 h. Moreover, N-PEG2 exhibited high transfection efficiency in comparison with lipofectamine® 2000 and low cytotoxicity. Therefore, the novel PEGylated cationic niosomes have the capability to develop as a promising potential carrier for DNA delivery.

Decaarginine-PEG-Artificial Lipid/DNA Complex for Gene Delivery: Nanostructure and Transfection Efficiency

2008 ◽  
Vol 8 (5) ◽  
pp. 2308-2315 ◽  
Author(s):  
Masahiko Furuhata ◽  
Radostin Danev ◽  
Kuniaki Nagayama ◽  
Yoshifumi Yamada ◽  
Hiroko Kawakami ◽  
...  
Keyword(s):  
Gene Delivery ◽  
Cellular Uptake ◽  
Plasmid Dna ◽  
Fluorescent Protein ◽  
Transfection Efficiency ◽  
Distribution Analysis ◽  
Lipid Concentration ◽  
Uptake Mechanism ◽  
Transmission Electron ◽  
Green Fluorescent

Oligoarginine conjugates are highly efficient vectors for the delivery of plasmid DNA into cells. Decaarginine-conjugated lipid (Arg10-PEG-lipid) was synthesized and the effects of Arg10-PEG-lipid concentration at a fixed DNA concentration on transfection efficiency and the structure of the complexes were studied below and above critical micelle concentration (CMC), and at the lipid nitrogen/DNA phosphate (N/P) ratio corresponding to transfection, respectively. Arg10-PEG-lipid at the concentration below CMC showed stronger interaction with DNA by fluorescence intensity distribution analysis, and significantly higher luciferase and green fluorescent protein expression than that above CMC. A phase-contrast cryo-transmission electron microscope (cryo-TEM) experiment showed that the morphology of the complexes depended on the N/P ratio. At a low N/P ratio corresponding to that in transfection at a lipid concentration below CMC, a net-like structure developed in which plasmid DNA was involved. A further increase in the N/P ratio, a large fibrous nanostructure of complexes, was also observed. Without DNA, these structures were not obtained. The cellular uptake mechanism of complexes using flow cytometry with inhibitors suggested that complexes with two different morphologies showed similar cellular uptake and uptake mechanism, macropinocytosis. Differences in transfection efficiency of the complexes may be explained by a large fibrous nanostructure inhibiting the cellular internalization of complexes or the release of DNA from macropinosomes into cytoplasm. Arg10-PEG-lipid/DNA complexes formed a favorable nanostructure for gene delivery, depending on the N/P ratio in water.

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