肝细胞靶向性半乳糖_聚乙二醇_聚乙烯亚胺纳米基因载体的合成及其理化特性

对聚乙二醇-聚乙烯亚胺进行半乳糖化修饰,可使其成为具有肝细胞靶向性的纳米基因载体半乳糖化聚乙二醇-聚乙烯亚胺[Galactosylated poly(ethylene glycol)-graft-polyethylenimine, Gal-PEG-PEI]。

中国组织工程研究与临床康复 第12卷 第36期 2008–09–02出版

Journal of Clinical Rehabilitative Tissue Engineering Research September 2, 2008 Vol.12, No.36

基础实验

肝细胞靶向性半乳糖-聚乙二醇-聚乙烯亚胺纳米基因载体的合成及其

☆ 理化特性**

聂常富1,刘春桃2,李 华1,张 彤1,杨 扬1,蔡常洁1,陆敏强1,帅心涛2,陈规划1

Synthesis and physicochemical characteristics of galactosylated poly(ethylene

glycol)-graft-polyethylenimine as a hepatocyte-targeting nano-gene carriers

Nie Chang-fu1, Liu Chun-tao2, Li Hua1, Zhang Tong1, Yang Yang1, Cai Chang-jie1, Lu Min-qiang1, Shuai Xin-tao2, Chen Gui-hua1

Abstract

BACKGROUND: Galactosylated poly(ethylene glycol)-graft-polyethylenimine (Gal-PEG-PEI) can be prepared as a nano-gene carrier of hepatocyte-targeting plasmid siRNA (psiRNA) through the glycosyl galactose of PEG-PEI.

OBJECTIVE: To synthesize Gal-PEG-PEI and study its physicochemical characteristics, so as to provide references for further study on the transfection.

DESIGN, TIME AND SETTING: The controlled experiments concerning about gene engineering were accomplished in Biomedical Engineering Centre of Sun Yat-sen University from October 2007 to June 2008. MATERIALS: The Gal-PEG-PEI was synthesized by two-step method.

METHODS: The physicochemical characteristics of the Gal-PEG-PEI/psiRNA nanoparticles were measured by nanoparticle-zeta electric potential instrument, transmission electron microscopy, gel retardation and ethidium bromide binding assay. Cytotoxicity of Gal-PEG-PEI was studied in cultured HepG2 cells by Cell Counting Kit-8 assay. The transfection experiments were performed with the Gal-PEG-PEI/psiRNA in HepG2 cells and the transfection efficiency was determined.

MAIN OUTCOME MEASURES: The particle sizes and zeta potentials of Gal-PEG-PEI/psiRNA; Encapsulation ability; Cytotoxicity results; Transfection efficiency in HepG2 cells.

RESULTS: The particle sizes of Gal-PEG-PEI/psiRNA decreased with increasing charge ratios of Gal-PEG-PEI to psiRNA and had a minimum value of 81.1 nm, while the zeta potentials were increased accordingly. Gal-PEG-PEI had a perfect ability to encapsulate psiRNA. Cytotoxicity of Gal-PEG-PEI was lower than that of PEI and transfection efficiency of Gal-PEG-PEI was higher than that of PEI without modification in HepG2 cells.

CONCLUSION: The Gal-PEG-PEI nano-gene carrier can be synthesized successfully, it displays perfect ability to hepatocyte-targeting and little cytotoxicity.

Nie CF, Liu CT, Li H, Zhang T, Yang Y, Cai CJ, Lu MQ, Shuai XT, Chen GH. Synthesis and physicochemical characteristics of galactosylated poly(ethylene glycol)-graft-polyethylenimine as a hepatocyte-targeting nano-gene carriers.Zhongguo Zuzhi Gongcheng Yanjiu yu Linchuang Kangfu 2008;12(36):7139-7142 [http://www.51wendang.com/zglckf/ejournal/upfiles/08-36/36k-7137(ps).pdf]

1

摘要

背景:对聚乙二醇-聚乙烯亚胺进行半乳糖化修饰,可使其成为具有肝细胞靶向性的纳米基因载体半乳糖化聚乙二醇-聚乙烯亚胺[Galactosylated poly(ethylene glycol)-graft-polyethylenimine, Gal-PEG-PEI]。

目的:合成Gal-PEG-PEI,考察其基本理化特征,为下一步的转染研究提供参考。

设计、时间及地点:对比观察基因工程实验,于2007-10/2008-06在中山大学生物医学工程中心完成。 材料:两步法合成Gal-PEG-PEI。

方法:应用纳米粒径-zeta电位仪、透射电镜、凝胶阻滞实验和EB结合实验测定Gal-PEG-PEI与psiRNA复合物的理化性质,CCK-8法检测其细胞毒性,纳米复合物转染HepG2细胞并观察其转染效率。

主要观察指标:Gal-PEG-PEI/psiRNA的粒径、zeta电位及形态;包覆效率;细胞毒性实验结果;肝脏靶向性转染结果。 结果:Gal-PEG-PEI/psiRNA复合物的粒径随N/P的增大而减小,最小粒径为81.1 nm,而zeta电位随Gal-PEG-PEI中氨基与psiRNA磷酸基比例的增大呈上升趋势;Gal-PEG-PEI对siRNA质粒有较好的包覆效率。Gal-PEG-PEI载体的细胞毒性小于聚乙烯亚胺而其转染效率高于聚乙烯亚胺。

结论:成功合成Gal-PEG-PEI纳米基因载体,表现出良好的肝细胞靶向性,且细胞毒性较小。 关键词:肝细胞靶向;半乳糖;纳米;非病毒基因载体;合成;特征

聂常富,刘春桃,李华,张彤,杨扬,蔡常洁,陆敏强,帅心涛,陈规划.肝细胞靶向性半乳糖-聚乙二醇-聚乙烯亚胺纳米基因载体的合成及其理化特性[J].中国组织工程研究与临床康复,2008,12(36):7139-7142 [http://www.51wendang.com/zglckf/ejournal/upfiles/08-36/36k-7137(ps).pdf]

>>本文导读<<

肝细胞靶向性半乳糖_聚乙二醇_聚乙烯亚胺纳米基因载体的合成及其理化特性

Liver

Transplantation Centre of the Third Affiliated Hospital, Sun Yat-sen University,

Guangzhou 510630, Guangdong Province, China; 2Biomedical Engineering Centre of Sun Yat-sen University,

Guangzhou 510080, Guangdong Province, China

Nie Chang-fu☆, Studying for

doctorate, Associate chief physician, Liver Transplantation Centre of the Third Affiliated Hospital, Sun Yat-sen University,

Guangzhou 510630, Guangdong Province, China

ncf1970@http://www.51wendang.com

Correspondence to: Chen Gui-hua, Doctor, Professor, Tutor of doctor, Liver Transplantation Centre of the Third Affiliated Hospital, Sun Yat-sen University,

Guangzhou 510630, Guangdong Province, China

chgh@http://www.51wendang.com

Supported by: the National Natural Science Foundation of China, No. 30571769*; the National Basic Research and Development Program of China (973 Program), No. 2003CB515507*

Received: 2008-07-03Accepted: 2008-08-04

ISSN 1673-8225 CN 21-1539/R CODEN: ZLKHAH

7139

肝细胞靶向性半乳糖_聚乙二醇_聚乙烯亚胺纳米基因载体的合成及其理化特性

肝细胞靶向性半乳糖_聚乙二醇_聚乙烯亚胺纳米基因载体的合成及其理化特性相关文档

最新文档

返回顶部