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• W2273157316 abstract "Introduction In recent years, growing attention have paid to interventional surgery for the treatment of cardiovascular and neurovascular diseases. The intravascular surgical techniques are largely relyed upon surgical devices such as balloon catheter, metallic stent, and embolic coil. The principal function of these devices includes physical reconstruction of vascular structure. The interventional surgery assisted with currently available devices is, however, associated with several problems including postoperative stenosis of lesion after in stenting, and recanalization in coil embolization. This paper presents our attempts to solve these problems by combining the conventional surgery with remote delivery of appropriate gene which serves to modulate tissue responses of the blood vessel. The final goal of this study is to develop a device that allows local delivery of a transfection vector encoding tissue growth factors such as bFGF and VEGF to surrounding tissue upon direct contact. As a model system, plasmid DNA carrying a reporter gene was immobilized onto the plainer surface of a thin gold film via the layer-by –layer adsorption technique [1]. Evaluation for gene transfection was carried out using in vitro cell culture in which mammalian cells were directly contacted with the DNA-modified surface. Experimental A thin gold layer with a thickness of 49 nm was deposited onto a glass plate by thermal evaporation. The gold-coated glass-plate was then immersed in 1 mM ethanol solution of 11 mercaptoundecanoic acid (HSC10H20COOH) for 24 h to form a self-assembled monolayer presenting carboxylic acid groups (COOH-SAM). The plasmid DNA encoding green fluorescent protein (GFP) gene was mixed with a cationic lipid (LipofectAMINE(TM)2000, Gibco) to prepare lipid-DNA complex baring a positive net charge. The complex was electrostatically adsorbed onto the COOH-SAM surface, and then the plasmid DNA with a negative net charge was overlaid. These procedures were repeated sequentially to obtain a substrate having a layer-by-layer structure of the lipid-DNA complex and DNA on the surface. The DNA-modified surface was characterized by the surface plasmon resonance (SPR) analysis and binding assay of a fluorescent probe, Picogreen®, having an affinity to DNA duplex. To verify the feasibility of gene transfers from the DNA-modified glass plate, HEK293 cells were directly seeded onto the plate at a cell. After 3-day culture, transfection efficiency was estimated by observing fluorescence emission due to the expression of GFP. Results It was probed by the surface plasmon resonance analysis that sequential adsorption of DNA and Lipid-DNA complex resulted in the formation of a Lipid/DNA complex layer. An increase in the thickness is also confirmed by quantity of incorporated DNA using of a DNA specific fluorescent probe. Fluorescent microphotograph of of HEK293 cells cultured on the DNA-modified glass plate demonstrated many cells exhibit fluorescent emission. This result indicates that the plasmid DNA was successfully transferred from the glass surface into cultured HEK 293 cells, leading to the intracellular expression of GFP." @default.
• W2273157316 created "2016-06-24" @default.
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• W2273157316 date "2002-06-01" @default.
• W2273157316 modified "2023-09-26" @default.
• W2273157316 title "Surface Modification for Interventional Remote Gene Delivery" @default.
• W2273157316 doi "https://doi.org/10.30047/jgmb.200206.0013" @default.
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