Functionalization of poly(amidoamine) dendrimers with hydrophobic chains for improved gene delivery in mesenchymal stem cells

Santos, José Lúıs; Oliveira, Hugo; Pandita, Deepti; Rodrigues, João; Pêgo, Ana Paula; Granja, Pedro L.; Tomás, Helena

doi:10.1016/j.jconrel.2010.01.034

Cited by 174 publications

(135 citation statements)

References 50 publications

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“…We chose the widely used dendrimer as a model to test the influence of the size and surface group as well as cytotoxicity and endocytosis on the hESC gene transfection. Our work is irrespective of those reports on dendrimer variable gene transfer efficacy in different cell types 18,20 and different from those observations of mere cytotoxicity but no influence on cellular differentiation of amino functionalized dendrimers for mesenchymal stem cells.…”

Section: ■ Discussioncontrasting

confidence: 99%

“…By contrast, G5, G4.5COOH, and G5OH were used to evaluate the impact of the end group on gene delivery upon previous observation that G5 is the best size for gene transduction for mesenchymal stem cells. 20 Dendrimer particle sizes and zeta potential were measured by dynamic light scattering (DLS) and electrophoretic light scattering (ELS) using a Zetasizer Nano-ZS, Malvern Instruments (Worcestershire, UK), in water or mTeSR medium at a concentration of 100 μM and pH 7.4. Dendrimers used for cellular study were dissolved in mTeSR medium at predetermined concentrations.…”

Section: ■ Experimental Sectionmentioning

confidence: 99%

“…(3) They have changeable surface functional groups besides amine, such as hydroxyl or carboxyl groups or even the arginine-glycine-aspartic acid (Arg-Gly-Asp, RGD) sequence. 20 …”

mentioning

confidence: 99%

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Improving the Gene Transfection in Human Embryonic Stem Cells: Balancing with Cytotoxicity and Pluripotent Maintenance

Luo

Lü

Pan

et al. 2016

ACS Appl. Mater. Interfaces

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Manipulation of genes in human embryonic stem cells (hESCs) is imperative for their highly potential applications; however, the transduction efficiency remains very low. Although existing evidence revealed the type, size, and zeta potential of vector affect gene transfection efficiency in cells, the systematic study in hESCs is scarce. In this study, using poly(amidoamine) (PAMAM) dendrimers ended with amine, hydroxyl, or carboxyl as model, we tested the influences of size and surface group as well as cytotoxicity and endocytosis on hESC gene transfection. We found that in culture medium of mTeSR the particle sizes of G5, G7, G4.5COOH, and G5OH were around 5 nm and G1 had a smaller size of 3.14 nm. G5 and G7 had a slight and significant positive zeta potential, respectively, whereas G1 was slightly negative, and G4.5COOH and G5OH were significantly negative. We demonstrated that only amine-terminated dendrimers accomplished gene transfection in hESCs, which is greater than that from Lipofectamine 2000 transfection. Ten micromolar G5 had the greatest efficiency and was better than 1000 μM G1. Only a low concentration (0.5 and 1 μM) of G7 realized gene delivery. Amine-ended dendrimers, especially with higher generations, were detrimental to the growth and pluripotent maintenance of hESCs. In contrast, similarly sized hydroxyl-and carboxyl-terminated dendrimers exerted much lower cytotoxicity, in which carboxyl-terminated dendrimer maintained pluripotency of hESCs. We also confirmed the endocytosis into and significant exocytosis from hESCs using FITC-labeled G5 dendrimer. These results suggested that careful considerations of size, concentration, and zeta potential, particularly the identity and position of groups, as well as minimized exocytosis in the design of a vector for hESC gene delivery are necessary, which helps to better design an effective vector in hESC gene transduction.

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Section: ■ Discussioncontrasting

confidence: 99%

Section: ■ Experimental Sectionmentioning

confidence: 99%

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Improving the Gene Transfection in Human Embryonic Stem Cells: Balancing with Cytotoxicity and Pluripotent Maintenance

Luo

Lü

Pan

et al. 2016

ACS Appl. Mater. Interfaces

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“…A possible answer is suggested by a balance of several factors influencing the transfection efficacy, such as chain length of the perfluoroaliphatic acid, number of conjugated ligands and residual surface charges, and degradability of the fluorinated dendrimer. This phenomenon is also observed in the gene transfection efficacies of dendrimers modified with aliphatic acid of different chain lengths 48 . The exact reason behind this question needs further investigations on the fluorinated dendrimers.…”

Section: Resultsmentioning

confidence: 71%

A fluorinated dendrimer achieves excellent gene transfection efficacy at extremely low nitrogen to phosphorus ratios

et al. 2014

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Polymers have shown great promise in the design of high efficient and low cytotoxic gene vectors. Here we synthesize fluorinated dendrimers for use as gene vectors. Fluorinated dendrimers achieve excellent gene transfection efficacy in several cell lines (higher than 90% in HEK293 and HeLa cells) at extremely low N/P ratios. These polymers show superior efficacy and biocompatibility compared with several commercial transfection reagents such as Lipofectamine 2000 and SuperFect. Fluorination enhances the cellular uptake of the dendrimer/DNA polyplexes and facilitates their endosomal escape. In addition, the fluorinated dendrimer shows excellent serum resistance and exhibits high gene transfection efficacy even in medium containing 50% FBS. The results suggest that fluorinated dendrimers are a new class of highly efficient gene vectors and fluorination is a promising strategy to design gene vectors without involving sophisticated syntheses.

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“…The addition of hydrophobic moieties to polycations has led to enhanced gene transfection in a variety of constructs such as polyethyleneimine (PEI), poly-l-lysine (PLL), PAMAM dendrimers, dendrons, and other similar structures (14,15). These modifications are thought to show their effects by a variety of mechanisms such as improved physical Ahad Mokhtarzadeh and Seyed Meghdad Tabatabai contributed equally to this work.…”

Section: Introductionmentioning

confidence: 99%

Preparation of Effective and Safe Gene Carriers by Grafting Alkyl Chains to Generation 5 Polypropyleneimine

et al. 2015

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Abstract. Gene therapy is a novel method to treat a variety of diseases including genetic disorders and cancer. Nonviral gene carriers have now gained considerable attention as gene carrier systems. Polyamidoamine (PAMAM) and polypropyleneimine (PPI) are the two most widely used denderimers in gene delivery studies. The aim of the current study was to investigate the effects of modification of generation 5 polypropyleneimine (G5 PPI) dendrimers with alkanoate groups as hydrophobic moieties on DNA transfection and cytotoxicity. Six, 10, and 16 carbon derivatives of bromoalkanoic acids were conjugated onto PPI with 10%, 30%, and 50% of surface amine grafting. Ethidium bromide exclusion assay results proved the ability of modified carriers to condense DNA. Transfection assay showed higher DNA delivery potential for 30% and 50% grafting with decanoate moieties compared to native G5 PPI and Superfect TM . 3-(4,5-Dimethylthiazol-2-yl)-2,5-di phenyltetrazolium bromide (MTT) and apoptosis experiments showed lower toxicity for modified carriers compared to unmodified PPI. The hemolytic effect of grafted carriers was not significantly different from G5 PPI. Size and zeta potential measurements revealed that polyplex size was less than 200 nm and electrical charges were in the range 14-25 mV. The hydrophobic modifications improved transfection activity and toxicity of G5 PPI without negatively affecting hemocompatibility. These modified carriers are therefore promising candidates for further in vivo investigations.

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Functionalization of poly(amidoamine) dendrimers with hydrophobic chains for improved gene delivery in mesenchymal stem cells

Cited by 174 publications

References 50 publications

Improving the Gene Transfection in Human Embryonic Stem Cells: Balancing with Cytotoxicity and Pluripotent Maintenance

Improving the Gene Transfection in Human Embryonic Stem Cells: Balancing with Cytotoxicity and Pluripotent Maintenance

A fluorinated dendrimer achieves excellent gene transfection efficacy at extremely low nitrogen to phosphorus ratios

Preparation of Effective and Safe Gene Carriers by Grafting Alkyl Chains to Generation 5 Polypropyleneimine

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