scholarly journals Binding Affinity, Cellular Uptake, and Subsequent Intracellular Trafficking of the Nano-Gene Vector P123-PEI-R13

2016 ◽  
Vol 2016 ◽  
pp. 1-8
Author(s):  
Yaguang Zhang ◽  
Hongmei Shu ◽  
Jing Hu ◽  
Min Zhang ◽  
Junweng Wu ◽  
...  
Keyword(s):  
Molecular Weight ◽  
Cellular Uptake ◽  
Intracellular Trafficking ◽  
Transfection Efficiency ◽  
Cross Linking ◽  
Low Molecular Weight ◽  
Gene Vector ◽  
Dna Complexes ◽  
Dynamic Source

A nano-gene vector PEI-P123-R13 was synthesized by cross-linking low molecular weight PEI with P123 and further coupling bifunctional peptide R13 to the polymer for targeting tumor and increasing cellular uptake. The binding assessment of R13 toαvβ3 positive cells was performed by HRP labeling. The internalization pathways of P123-PEI-R13/DNA complexes were investigated based on the effect of specific endocytic inhibitors on transfection efficiency. The mechanism of intracellular trafficking was investigated based on the effect of endosome-lysosome acidification inhibitors, cytoskeleton, and dynein inhibitors on transfection efficiency. The results indicated that the bifunctional peptide R13 had the ability of binding toαvβ3 positive cellsin vitro. The modification of P123-PEI-R13 with R13 made it display new property of internalization. P123-PEI-R13/DNA complexes were conducted simultaneously via clathrin-mediated endocytosis, caveolin-mediated endocytosis, macropinocytosis, and possible energy-independent route. After internalization, P123-PEI-R13/DNA complexes could escape from the endosome-lysosome system because of its acidification and further took microtubule as the track and dynein as the dynamic source to be transported toward the microtubule (+) end, to wit nucleus, under the action of microfilament, and with the aid of intermediate filament.

Low Molecular Weight Branched Polyethylenimine-Graft-Lentinan as a Novel Synthetic Gene Carrier

2013 ◽  
Vol 815 ◽  
pp. 293-298
Author(s):  
Li Jing Min ◽  
Jing Fen Li
Keyword(s):  
Molecular Weight ◽  
Gel Electrophoresis ◽  
Transfection Efficiency ◽  
Gene Transfection ◽  
Water Soluble ◽  
Low Molecular Weight ◽  
Dna Migration ◽  
Branched Polyethylenimine ◽  
Soluble Polysaccharide

LMW Branched Polyethylenimine-graft-lentinan was synthesized by performing crosslinker CDI in the presence of water-soluble Polysaccharide (Mw 70,000) and polyethylenimine (PEI, Mw 600). The chemistry of the PEI-g-lentinan obtained were characterized. The results indicated that the amines of lentinan were grafted with PEI. Gel electrophoresis showed that DNA migration was retarded completely at a N/P ratio of 10/1, indicating good DNA condensation capability of PEI-g-lentinan. The cytotoxicity of PEI-g-lentinan was evaluated, and the results reflected that PEI-g-lentinan had a lower cytotoxicity than PEI (25 K). Gene transfection efficiency of PEI-g-chitosan in cos-7 cells was determined. Remarkably,PEI-g-lentinan showed a higher transfection efficiency than that of PEI (25 K) in vitro.

scholarly journals Synergistic Effect of Low Molecular Weight Polyethylenimine and Polyethylene Glycol Components in Dynamic Nonviral Vector Structure, Toxicity, and Transfection Efficiency

Molecules ◽  
2019 ◽  
Vol 24 (8) ◽  
pp. 1460 ◽  
Author(s):  
Bogdan Florin Craciun ◽  
Gabriela Gavril ◽  
Dragos Peptanariu ◽  
Laura Elena Ursu ◽  
Lilia Clima ◽  
...  
Keyword(s):  
Molecular Weight ◽  
Nucleic Acid ◽  
Transfection Efficiency ◽  
Morphological Data ◽  
Low Molecular Weight ◽  
Nonviral Vectors ◽  
Main Challenge ◽  
Vector Structure ◽  
Imine Bond

When studying polyethylenimine derivatives as nonviral vectors for gene delivery, among the important issues to be addressed are high toxicity, low transfection efficiency, and nucleic acid polyplex condensation. The molecular weight of polyethylenimine, PEGylation, biocompatibility and, also, supramolecular structure of potential carrier can all influence the nucleic acid condensation behavior, polyplex size, and transfection efficiency. The main challenge in building an efficient carrier is to find a correlation between the constituent components, as well as the synergy between them, to transport and to release, in a specific manner, different molecules of interest. In the present study, we investigated the synergy between components in dynamic combinatorial frameworks formed by connecting PEGylated squalene, poly-(ethyleneglycol)-bis(3-aminopropyl) and low molecular weight polyethylenimine components to 1,3,5-benzenetrialdehyde, via reversible imine bond, applying a dynamic combinatorial chemistry approach. We report comparative structural and morphological data, DNA binding affinity, toxicity and transfection efficiency concerning the ratio of polyethylenimine and presence or absence of poly-(ethyleneglycol)-bis(3-aminopropyl) in composition of dynamic combinatorial frameworks. In vitro biological assessments have revealed the fact that nonviral vectors containing poly-(ethyleneglycol)-bis(3-aminopropyl) and the lowest amount of polyethylenimine have significant transfection efficiency at N/P 50 ratio and display insignificant cytotoxicity on the HeLa cell line.

scholarly journals A Low-Molecular-Weight Polyethylenimine/pDNA-VEGF Polyplex System Constructed in a One-Pot Manner for Hindlimb Ischemia Therapy

Pharmaceutics ◽  
2019 ◽  
Vol 11 (4) ◽  
pp. 171 ◽  
Author(s):  
Xiaoshuang Guo ◽  
Zihan Yuan ◽  
Yang Xu ◽  
Xiaotian Zhao ◽  
Zhiwei Fang ◽  
...  
Keyword(s):  
Molecular Weight ◽  
Gene Delivery ◽  
High Efficiency ◽  
Transfection Efficiency ◽  
Low Molecular Weight ◽  
Physical Interaction ◽  
Hindlimb Ischemia ◽  
One Pot

Peripheral arterial disease (PAD) is often characterized by continued reduction in blood flow supply to limbs. Advanced therapeutic strategies like gene therapy could potentially be applied to limb ischemia therapy. However, developing a gene delivery system with low toxicity and high efficiency remains a great challenge. In this study, a one-pot construction was used to integrate vector synthesis and polyplex fabrication simultaneously in a simple and robust manner. We fabricated an interpenetrating gene delivery network through the physical interaction between low-molecular-weight polyethylenimine (PEI 1.8 kDa) and plasmid DNA (pDNA) and the chemical bonding between PEI and glutaraldehyde (GA), which was named the glutaraldehydelinked-branched PEI (GPEI) polyplex. The final GPEI polyplex system was pH-responsive and biodegradable due to the imine linkage and it could successfully deliver desired vascular endothelial growth factor (VEGF) pDNA. Compared with PEI (25 kDa)/pDNA polyplexes, GPEI polyplexes showed lower cytotoxicity and higher transfection efficiency both in vitro and in vivo. In addition, we demonstrated that GPEI polyplexes could efficiently promote the formation of new capillaries in vivo, which may provide a practicable strategy for clinical hindlimb ischemia therapy in the future.

scholarly journals Synthesis and Properties of Low-Molecular-Weight PEI-Based Lipopolymers for Delivery of DNA

Polymers ◽  
2018 ◽  
Vol 10 (10) ◽  
pp. 1060 ◽  
Author(s):  
Miao-Miao Xun ◽  
Zheng Huang ◽  
Ya-Ping Xiao ◽  
Yan-Hong Liu ◽  
Ji Zhang ◽  
...  
Keyword(s):  
Drug Delivery ◽  
Molecular Weight ◽  
Transfection Efficiency ◽  
Dna Delivery ◽  
Low Molecular Weight ◽  
Zeta Potentials ◽  
Poor Efficacy ◽  
Mcf 7

Rapid enzymatic degradation and fragmentation during DNA administration can result in limited gene expression, and consequently, poor efficacy. It is necessary to use novel vectors for DNA delivery. Herein, we aimed to design useful carriers for enhancing transfection efficiency (TE). These lipopolymers were prepared through Michael addition reactions from low-molecular-weight (LMW) polyethyleneimine (PEI) and linkers with three kinds of steroids. Agarose gel electrophoresis assay results displayed that the three lipopolymers could condense plasmid DNA well, and the formed polyplexes had appropriate sizes around 200–300 nm, and zeta potentials of about +25–40 mV. The results of in vitro experiments using HeLa, HEK293, and MCF-7 cells showed that these lipopolymers present higher TE than 25-kDa PEI, both in the absence and presence of 10% serum. Flow cytometry and confocal microscopy studies also demonstrated that these lipopolymer/DNA complexes present higher cellular uptake and intracellular distribution. The measurement of critical micelle concentration (CMC) revealed that these lipopolymers could form micelles, which are suited for drug delivery. All results suggest that the three materials may serve as hopeful candidates for gene and drug delivery in future in vivo applications.

Cellular Uptake and Transfection Efficiency of Plasmid DNA Using Low Molecular Weight Polyethylenimine

2004 ◽  
Vol 34 (4) ◽  
pp. 263-267
Author(s):  
Gil-Jae Jeong ◽  
Kui-Lye Park ◽  
Ji-Young Shin ◽  
Han-Gon Choi ◽  
Yu-Kyoung Oh
Keyword(s):  
Molecular Weight ◽  
Cellular Uptake ◽  
Plasmid Dna ◽  
Transfection Efficiency ◽  
Low Molecular Weight

The low-molecular-weight phosphotyrosine phosphatase is a negative regulator of FcγRIIA-mediated cell activation

Blood ◽  
2007 ◽  
Vol 110 (6) ◽  
pp. 1871-1878 ◽  
Author(s):  
Francesca Mancini ◽  
Stefania Rigacci ◽  
Andrea Berti ◽  
Cesare Balduini ◽  
Mauro Torti
Keyword(s):  
Molecular Weight ◽  
Cell Activation ◽  
Human Platelets ◽  
Negative Regulator ◽  
Cross Linking ◽  
Low Molecular Weight ◽  
Phosphotyrosine Phosphatase ◽  
Affinity Receptor ◽  
Activation Motif

Abstract Activation of human platelets by cross-linking of the low-affinity receptor for immunoglobulin G (FcγRIIA) is initiated by Src kinase–mediated phosphorylation of the immunoreceptor tyrosine–based activation motif (ITAM) within the receptor, but the identity of the enzyme responsible for its dephosphorylation and inactivation is unknown. Here we report that the 18-kDa low-molecular-weight phosphotyrosine phosphatase (LMW-PTP) is expressed in human platelets and undergoes subcellular redistribution upon FcγRIIA cross-linking. In vitro, LMW-PTP was found to efficiently dephosphorylate activated FcγRIIA and LAT, but not Syk or phospholipase Cγ2. In the megakaryocytic cell line DAMI, antibody-induced phosphorylation of FcγRIIA was rapid and transient. The late dephosphorylation of FcγRIIA was dramatically delayed upon reduction of LMW-PTP expression by siRNA. Strikingly, overexpression of LMW-PTP resulted in the inhibition of antibody-induced phosphorylation of FcγRIIA, and caused a more rapid dephosphorylation. In addition, overexpression of LMW-PTP inhibited activation of Syk downstream of FcγRIIA and reduced intracellular Ca2+ mobilization. These results demonstrate that LMW-PTP is responsible for FcγRIIA dephosphorylation, and is implicated in the down-regulation of cell activation mediated by this ITAM-bearing immunoreceptor.

Effects of Unfractionated and Low Molecular Weight Heparins on Platelet Thromboxane Biosynthesis “In Vivo”

1994 ◽  
Vol 72 (06) ◽  
pp. 942-946 ◽  
Author(s):  
Raffaele Landolfi ◽  
Erica De Candia ◽  
Bianca Rocca ◽  
Giovanni Ciabattoni ◽  
Armando Antinori ◽  
...  
Keyword(s):  
Molecular Weight ◽  
Unfractionated Heparin ◽  
Low Molecular Weight Heparin ◽  
Primary Source ◽  
In Vivo Studies ◽  
Low Molecular Weight ◽  
Heparin Administration ◽  
To Receive

SummarySeveral “in vitro” and “in vivo” studies indicate that heparin administration may affect platelet function. In this study we investigated the effects of prophylactic heparin on thromboxane (Tx)A2 biosynthesis “in vivo”, as assessed by the urinary excretion of major enzymatic metabolites 11-dehydro-TxB2 and 2,3-dinor-TxB2. Twenty-four patients who were candidates for cholecystectomy because of uncomplicated lithiasis were randomly assigned to receive placebo, unfractionated heparin, low molecular weight heparin or unfractionaed heparin plus 100 mg aspirin. Measurements of daily excretion of Tx metabolites were performed before and during the treatment. In the groups assigned to placebo and to low molecular weight heparin there was no statistically significant modification of Tx metabolite excretion while patients receiving unfractionated heparin had a significant increase of both metabolites (11-dehydro-TxB2: 3844 ± 1388 vs 2092 ±777, p <0.05; 2,3-dinor-TxB2: 2737 ± 808 vs 1535 ± 771 pg/mg creatinine, p <0.05). In patients randomized to receive low-dose aspirin plus unfractionated heparin the excretion of the two metabolites was largely suppressed thus suggesting that platelets are the primary source of enhanced thromboxane biosynthesis associated with heparin administration. These data indicate that unfractionated heparin causes platelet activation “in vivo” and suggest that the use of low molecular weight heparin may avoid this complication.

The Effect of Dextran of Various Molecular Weight on the Coagulation in Dogs

1961 ◽  
Vol 06 (01) ◽  
pp. 015-024 ◽  
Author(s):  
Sven Erik Bergentz ◽  
Oddvar Eiken ◽  
Inga Marie Nilsson
Keyword(s):  
Molecular Weight ◽  
Body Weight ◽  
High Molecular Weight ◽  
Bleeding Time ◽  
Factor Vii ◽  
Low Molecular Weight ◽  
Factor V ◽  
Coagulation Mechanism ◽  
Coagulation Defects

Summary1. Infusions of low molecular weight dextran (Mw = 42 000) to dogs in doses of 1—1.5 g per kg body weight did not produce any significant changes in the coagulation mechanism.2. Infusions of high molecular weight dextran (Mw = 1 000 000) to dogs in doses of 1—1.5 g per kg body weight produced severe defects in the coagulation mechanism, namely prolongation of bleeding time and coagulation time, thrombocytopenia, pathological prothrombin consumption, decrease of fibrinogen, prothrombin and factor VII, factor V and AHG.3. Heparin treatment of the dogs was found to prevent the decrease of fibrinogen, prothrombin and factor VII, and factor V otherwise occurring after injection of high molecular weight dextran. Thrombocytopenia was not prevented.4. In in vitro experiments an interaction between fibrinogen and dextran of high and low molecular weight was found to take place in systems comprising pure fibrinogen. No such interaction occurred in the presence of plasma.5. It is concluded that the coagulation defects induced by infusions of high molecular weight dextran are due to intravascular coagulation.

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