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载有血小板反应蛋白-1 基因的聚乙二醇化壳聚糖和聚乙烯亚胺聚合物的制剂及体外特性研究。

Formulation and in vitro characterization of PEGylated chitosan and polyethylene imine polymers with thrombospondin-I gene bearing pDNA.

机构信息

Department of Pharmaceutical Technology, Faculty of Pharmacy, Ankara University, Ankara, Turkey.

出版信息

J Biomed Mater Res B Appl Biomater. 2012 May;100(4):984-92. doi: 10.1002/jbm.b.32661. Epub 2012 Jan 25.

DOI:10.1002/jbm.b.32661
PMID:22279036
Abstract

An ideal gene carrier is required both in safety and efficiency for transfection. We examined the use of water soluble chitosan and polyethyleneimine as a carrier for anti-angiogenic protein, TSP-1 coded, in gene delivery. The aim of this study was to synthesize and characterize polyethylene glycol conjugated cationic polymers to increase anti-angiogenic gene transfection and reduce possible cytotoxicity. Gel electrophoresis study showed strong DNA binding ability of modified cationic polymers. Also structural properties of pegylated polymers were confirmed by (1)H-NMR. We investigated in vitro properties of PEG conjugated and coated particles which were observed between 145 and 250 nm with the positive zeta potential value. In addition, the chitosan-based DNA complexes did not induce remarkable cytotoxicity against MCF-7 cells. Due to low cytotoxicity, we observed high transfection efficiency at chitosan-based formulations compared with PEI ones. Although transfection studies carried on in vitro conditions, we measured slight increases at transfection with PEGylation. PEG-conjugated chitosan formulations can be a promising candidate due to its efficiency in condensing and transfection of pDNA, its low cytotoxicty and comparatively high encapsulation degree.

摘要

对于转染来说,理想的基因载体既要求安全又要求高效。我们研究了水溶性壳聚糖和聚乙烯亚胺作为编码抗血管生成蛋白 TSP-1 的载体在基因传递中的应用。本研究的目的是合成和表征聚乙二醇修饰的阳离子聚合物,以提高抗血管生成基因的转染效率并降低可能的细胞毒性。凝胶电泳研究表明,修饰后的阳离子聚合物具有很强的 DNA 结合能力。此外,(1)H-NMR 也证实了聚乙二醇化聚合物的结构特性。我们研究了 PEG 修饰和涂层颗粒的体外性质,这些颗粒的粒径在 145 到 250nm 之间,具有正的zeta 电位值。此外,壳聚糖基 DNA 复合物对 MCF-7 细胞没有明显的细胞毒性。由于低细胞毒性,与 PEI 相比,我们观察到壳聚糖基制剂具有更高的转染效率。尽管转染研究是在体外条件下进行的,但我们发现 PEG 化后转染效率略有提高。由于其在凝聚和转染 pDNA 方面的效率、低细胞毒性和相对较高的包封率,PEG 修饰的壳聚糖制剂可能是一种很有前途的候选物。

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