Papers by Keyword: Gene Delivery

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Authors: Dhananjay Jere, Tae Hee Kim, Rohidas B. Arote, Hu Lin Jiang, Myung Haing Cho, Jae Woon Nah, Chong Su Cho
Abstract: Vectors are vital aspect of gene delivery system which decides the success of gene therapy. Efficient transfection with minimum or no toxicity, are two principal aims of any gene delivery system. In this our study, we rationally developed biodegradable water soluble poly(ßamino ester) (PAE) based on spermine (SPR) and poly (ethylene glycol) (PEG), by Michael-type addition reaction and further studied for its potential as a gene carrier. Confirmation of synthesized PAE was done by proton NMR spectroscopy. In gel retardation assay, the PAEs have shown good DNA binding ability over wide range of polyplexes. The addition of PEG over SPR resulted in a novel PAE with higher degree of safety and transfection efficiency as compared with polyethylenimine 25K (PEI) when studied in 293T human kidney carcinoma cells.
Authors: Mohammad A. Jafar Mazumder, Md. Hasan Zahir, Sharif F. Zaman
Abstract: Gene therapy is a widespread and promising treatment of many diseases resulting from genetic disorders, infections and cancer. The feasibility of the gene therapy is mainly depends on the development of appropriate method and suitable vectors. For an efficient gene delivery, it is very important to use a carrier that is easy to produce, stable, non-oncogenic and non-immunogenic. Currently most of the vectors actually suffer from many problems. Therefore, the ideal gene therapy delivery system should be developed that can be easily used for highly efficient delivery and able to maintain long-term gene expression, and can be applicable to basic research as well as clinical settings. This article provides a brief over view on the concept and aim of gene delivery, the different gene delivery systems and use of different materials as a carrier in the area of gene therapy.
Authors: Cun Xian Song, Man Yan Wang, Jing Yang, Xu Jin, Lin Mei, Xigang Leng, Hong Fan Sun, R.J. Levy
Authors: Wanlop Weecharangsan, Orapan Paecharoenchai, Nattisa Niyomtham, Praneet Opanasopit, Boon-ek Yingyongnarongkul, Robert J. Lee
Abstract: 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.
Authors: Naznin Sultana, Nobuhiro Nakamura, Shigehisa Hirose, Koichi Kutsuzawa, Toshihiro Akaike, Kakon Nag
Abstract: Heart development is a precisely harmonized process of cellular proliferation, migration, differentiation, and integrated morphogenetic interactions, and therefore it is extremely vulnerable to developmental defects that cause congenital heart diseases (CHD). One of the major causes of CHD has been shown to be the mutations in key cardiac channel-forming proteins namely, connexins (Cxs). Cxs are tetra-spanning transmembrane proteins that form gap junction channels and hemichannels on cellular membrane. They allow passage of small molecules or ions between adjacent cells or between cells and the extracellular environment. Studies have revealed that the spatiotemporal expression of Cxs mainly, Cx31.9, Cx40, Cx43, and Cx45 is essentially involved in early developmental events, morphogenetic transformations, maturation, and functional significance of heart. Our lab and others have shown that mutations in gap junction proteins could result in impaired trafficking, misfolding, and improper channel function of these proteins. It has also been shown that differential expressions of cardiac Cxs are associated with pathophysiological conditions of heart. Collectively, these conditions are coupled with abrogated or modified functionality of relevant channels in cardiac tissue, which are associated with many pathological situations, including CHD. Since CHD are a major cause of morbidity, therefore recovery of such kind of heart defects associated with Cxs is extremely important but remains highly challenging. In this review, we will summarize the role of Cxs in development, morphogenesis, maturation, normal function, and pathology of heart, and propose possible bioengineering techniques to recover defects in cardiac tissues related to the modified functions of Cxs.
Authors: Ke Feng Ren, Jian Ji, Jia Cong Shen
Abstract: Films composed of alternating layers of protamine and DNA were constructed using the layer-by-layer method on quartz and subsequently studied the enzymic degradation in vitro. UV-visible spectrometry measurement indicated the uniform assembly of Protamine/DNA multilayer films. UV-visible spectrometry and fluorescence spectrometry results revealed that the Protamine/DNA multilayer films were in vitro enzymic biodegradable. The novel biodegradable multilayer of Protamine/DNA may have great potential for gene therapy applications in tissue engineering, medical implant etc.
Authors: Ayako Oyane, Hiroko Araki, Yu Sogo, Atsuo Ito, Hideo Tsurushima
Abstract: A surface-mediated gene transfer system using DNA-calcium phosphate (CaP) composite layers (D-CaP layers) would be useful in tissue engineeing. In previous studies, D-CaP layers were fabricated in supersaturated CaP solutions prepared using chemical reagents. In this study, a so-called RKM solution prepared using clinically approved infusion fluids was employed as a supersaturated CaP solution. A D-CaP layer consisting of submicron spherical particles was successfully fabricated on a polystyrene substrate by immersing the substrate in the RKM solution for 24 h. When the immersion period was prolonged from 24 to 72 h, amount of CaP and DNA on the substrate increased. However, the gene transfer capability of the D-CaP layer for the CHO-K1 cells was kept unchanged irrespective of the immersion period. In the RKM solution process, immersion period of 24 h was found to be long enough for gene transfer application of the D-CaP layer. More importantly, the D-CaP layer fabricated by the RKM solution process exhibited a significantly higher gene transfer capability than our previous D-CaP layer fabricated in the conventional CaP solution with the same DNA concentration. The RKM solution process for the fabrication of D-CaP layers was found to be advantageous to the previous process in terms of not only safety but the layers gene transfer capability.
Authors: Orapan Paecharoenchai, Tittaya Suksamran, Tanasait Ngawhirunpat, Theerasak Rojanarata, Praneet Opanasopit
Abstract: Chitosan nanoparticles were prepared by crosslinking chitosan (CS) with tripolyphosphate (TPP) solution using electrohydrodynamic spraying technique. The effects of CS and TPP concentration as well as electrical potential on particle size and shape were investigated. Appropriated formulations for preparing nanoparticles were chosen to encapsulate DNA. In vitro evaluation of the obtained nanoparticles as gene carrier such as entrapment efficiency and DNA release was performed. The results showed that 2 mg/ml TPP was dropped at 10 kV into 1 mg/ml CS (MW 20 kDa (F1) and 200 kDa (F2)) yielded the spherical shape and small particles of 227.67 and 240.33 nm, respectively. In DNA entrapment study, all formulations were tested by altering DNA loading to 10, 25 and 50 mg/g of CS. The results revealed that F1 with initial DNA 10 mg/g of CS showed the highest entrapment efficiency of 95.31%. While F2 with initial DNA of 25 mg/g of CS showed the highest entrapment efficiency of 89.16%. The DNA release study from CS nanoparticles indicated that the increasing of DNA amount slowed down the release rate. F1 and F2 with the initial DNA of 10 mg/g of CS had faster release rate than those with 25 and 50 mg/g of CS. It can be concluded that F1 yielded the nanoparticles with the smallest size, high DNA entrapment efficiency and enabled DNA sustained release.
Authors: Qing Wen Guan, Min Wang
Abstract: Gene therapy has great potential in offering highly promising treatments for cancer. Polymer-metal hybrid nanoparticles (NPs) are good candidates as gene delivery vehicles due to their unique properties and facile functionalization. The polymer component in hybrid NPs can provide accurate cancer cell targeting and high DNA binding ability while the metallic component can provide imaging functions for the nanodevices. In the present study, hybrid NPs comprising an Au-Ag bimetallic core and a folic acid-chitosan shell (Au-Ag@CS-FA) were fabricated. The structure and relevant properties of Au-Ag@CS-FA NPs were subsequently studied using a variety of techniques,like scanning electron microscope (SEM), Fourier transform infrared spectroscopy (FTIR), transmission electron microscope (TEM) and UV-visible spectra. Their DNA binding ability was also assessed. Results showed that Au-Ag@CS-FA NPs possessed properties that can make them excellent gene delivery vehicles.
Authors: Leng Nie, Li Zeng Gao, Xi Yun Yan, Tai Hong Wang
Abstract: Amino-modified tetrapod-like ZnO nanostructures were tried as novel carriers for mammalian cell transfections. The nanostructures consisted of four needle-shaped tetrahedrally arranged legs connected at the center. After silica coating and amino modification, ZnO nanostructures complexes bound plasmid DNA through electrostatic interactions in aqueous solution. When mixed with cells, DNA-nanostructures attached easily onto cell membranes and entered the cells for gene expressions. Due to high positive charge densities on surfaces and needle-shaped tetrahedral structures, functionalized ZnO used as carriers for cell transfections with both high transfection efficiency and little cytotoxicity. And a possible transfection machamism was proposed in this report.
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