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基于星型聚合物的纳米层,其中固定有聚阳离子星和 DNA 的复合物,用于沉积基因传递和完整转染细胞的回收。

Star polymer-based nanolayers with immobilized complexes of polycationic stars and DNA for deposition gene delivery and recovery of intact transfected cells.

机构信息

Centre of Polymer and Carbon Materials, Polish Academy of Sciences, M. Curie-Sklodowskiej 34, 41-819 Zabrze, Poland.

Department of Molecular Biology and Genetics, Faculty of Medical Sciences in Katowice, Medical University of Silesia, Katowice, Medykow 18 Street, 40-752 Katowice, Poland.

出版信息

Int J Pharm. 2020 Nov 15;589:119823. doi: 10.1016/j.ijpharm.2020.119823. Epub 2020 Aug 28.

Abstract

We designed a novel thermoresponsive system of nanolayers composed of star poly[oligo(ethylene glycol) methacrylate]s (S-POEGMA) covalently bonded to a solid support and covered with polyplexes of cationic star polymers and plasmid DNA (pDNA). S-POEGMA stars were attached to the solid support via a UV-mediated "grafting to" method. To the best of our knowledge, for the first time, the conformational changes of obtained star nanolayers, occurring with changes in temperature, were studied using a quartz crystal microbalance technique. Next, the polyplexes of star poly[N,N'-dimethylaminoethyl methacrylate-ran-di(ethylene glycol) methacrylate] (S-P(DMAEMA-DEGMA)) with pDNA, exhibiting a phase transition temperature (T) in culture medium DMEM, were deposited on S-POEGMA layers when the temperature increased above the T of polyplex. The thermoresponsivity of the system was then the main mechanism for controlling the adhesion, proliferation, transfection and detachment of HT-1080 cells. The nanolayers promoted the effective cell culture and delivered nucleic acids into cells, with a transfection efficiency several times higher than that of the control. The detachment of the transfected cells was regulated only by the change of temperature. The studies demonstrated that we obtained a novel and effective system, based on a star polymer architecture, useful for gene delivery and tissue engineering applications.

摘要

我们设计了一种新型的纳米层热响应体系,由共价键合到固体支撑物上的星型聚[低聚(乙二醇)甲基丙烯酸酯](S-POEGMA)和阳离子星型聚合物与质粒 DNA(pDNA)的聚集体组成。S-POEGMA 星型物通过紫外介导的“接枝到”方法连接到固体支撑物上。据我们所知,这是首次使用石英晶体微天平技术研究了所得星型纳米层在温度变化时发生的构象变化。接下来,当温度升高超过聚集体的相转变温度(T)时,在含有 pDNA 的星型聚[N,N'-二甲氨基乙基甲基丙烯酸酯-ran-二(乙二醇)甲基丙烯酸酯](S-P(DMAEMA-DEGMA))聚集体上沉积 S-POEGMA 层。然后,该体系的热响应性成为控制 HT-1080 细胞粘附、增殖、转染和脱落的主要机制。该纳米层促进了有效的细胞培养,并将核酸递送到细胞中,其转染效率比对照高几倍。仅通过温度变化即可调节转染细胞的脱落。研究表明,我们获得了一种基于星型聚合物结构的新型有效体系,可用于基因传递和组织工程应用。

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