利用電場控制導電性高分子以進行基因於聚電解質多層膜的組裝與釋放;The assembly and disassembly of polyelectrolyte multilayer regulated by electrical control using conductive polymer

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题名:	利用電場控制導電性高分子以進行基因於聚電解質多層膜的組裝與釋放;The assembly and disassembly of polyelectrolyte multilayer regulated by electrical control using conductive polymer
作者:	胡威文
贡献者:	國立中央大學化學工程與材料工程學系
关键词:	化學工程;生物技術(醫)
日期:	2013-12-01
上传时间:	2014-03-17 11:30:55 (UTC+8)
出版者:	行政院國家科學委員會
摘要:	研究期間：10208~10307;In this study, we would like to develop an electrical control gene delivery using conductive polymer as substrates. DNA will be incorporated with chitosan as polyelectrolyte multilayers (PEMs) using layer-by-layer (LbL) techniques. Because the electrostatic interaction is the main driving force to immobilize polycation and polyanion on surfaces, the polarity of substrates should be able to be controlled through adjusting external electrical fields. The electrical effects on the assembly and disassembly of PEMs will be both studied. Conductive polymer, polypyrrole (PPy) films, will be synthesized using different concentrations of monomer and initiator. Their morphology, thickness, and electrical properties will be analyzed. To regulate DNA assembly, vertical electrical fields will be applied. DNA loading capacities, their protection ability to DNA, and cytotoxicity will be examined to ensure that the potential may significantly improve DNA incorporation without eliciting negatively effects on surface cells. On the other hand, we will also develop an electrical control of PEMs disassembly to regulate DNA release from substrates. Horizontal electrodes will be placed in two ends of PPy films. Different electrical stimulation will be applied to study their effects on the unstablization of PEMs. Finally, we will apply this electrical control system to in situ deliver osteogenic factor gene, BMP-7, to placenta-derived multipotent cells (PDMCs) on PPy films. The biocompatibility and cytotoxicity of this gene delivery system will be assessed by MTT and LDH assays, respectively. The internalized BMP-7 gene will be quantified by quantitative real-time PCR, and the expressed BMP-7 will be examined by ELISA. The time sequence of BMP-7 expression will be determined with the levels of mRNA encoding osteoblast differentiation markers. The mineralization level will also be assessed to determine the effect of DNA immobilization strategy on mineral tissue regeneration. With or without applying electrical field will be performed to distinguish the effects of gene delivery and electrical stimulation on cell differentiation. Through these assays we may determine the extent to which this in situ gene delivery method could temporally control cell differentiation and tissue regeneration in specific period. We believe that this interdisciplinary and perspective project should benefit the field of regenerative gene therapy, expand the depth and extent of the tissue engineering research, and increase the visibility of our country in sciences.
關聯:	財團法人國家實驗研究院科技政策研究與資訊中心
显示于类别:	[化學工程與材料工程學系 ] 研究計畫

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