聚乙烯亚胺改性的双介孔氧化硅基因载体构建  

Construction of Polyethyleneimine-modified Dual-mesoporous Silica Gene Carrier

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作  者:方美蓉 秦利梅 贾晓博[2] 李永生[2] 牛德超 胡泽岚 FANG Mei-Rong;QIN Li-Mei;JIA Xiao-Bo;LI Yong-Sheng;NIU De-Chao;HU Ze-Lan(Shanghai Key Laboratory of New Drug Design,School of Pharmacy,East China University of Science and Technology,Shanghai 200237,China;School of Materials Science and Engineering,East China University of Science and Technology,Shanghai 200237,China)

机构地区:[1]华东理工大学药学院上海新药设计重点实验室,上海200237 [2]华东理工大学材料科学与工程学院,上海200237

出  处:《无机材料学报》2020年第2期187-192,共6页Journal of Inorganic Materials

基  金:国家自然科学基金(51572083,81772689);上海市科技启明星计划(16QA1401300)~~

摘  要:为了开发一种新型的非病毒无机基因载体,采用3-氨丙基三乙氧基硅烷(APTMS)和聚乙烯亚胺(PEI)对嵌入型双介孔氧化硅球(EDMSNs)进行改性,分别得到EDMSNs-NH2和EDMSNs-PEI,并比较了两种载体结合和保护pCMV-EGFP-N1质粒(pDNA)的能力及细胞转染性能。利用透射电镜、动态光散射及氮气吸附-脱附实验对材料的颗粒形态,动力学粒径,Zeta电位及孔结构参数进行表征。结果显示,EDMSNs-NH2和EDMSNs-PEI均表现出明显的双介孔结构,形貌为规整的球形且平均动力学粒径分别为343.2和338.9 nm,表面电位分别为+18和+43 mV。琼脂糖凝胶电泳、CCK-8法及荧光显微镜结果表明,EDMSNs-PEI对pDNA的担载量为8%,远高于EDMSNs-NH2(1%)。与PEI和lipofectamine2000相比,EDMSNs-PEI载体展示出更低的细胞毒性。EDMSNs-PEI/pDNA质量比为33:1时,EDMSNs-PEI/pDNA对293T细胞的转染效率在72 h达到最大值。因此,与EDMSNs-NH2相比,EDMSNs-PEI具有更高的正电位及pDNA担载量,可作为一类有前景的非病毒无机类基因载体用于重大疾病如恶性胶质瘤的治疗。To develop a new kind of non-viral inorganic gene vector,embedded dual-mesoporous silica spheres(EDMSNs)was modified with polyethyleneimine(PEI)or 3-aminopropyltriethoxysilane(APTMS).Then gene loading and transfection ability of the obtained EDMSNs-PEI and EDMSNs-NH2 was tested by choosing the pCMV-EGFP-N1 plasmids(pDNA)as a gene model.Particle morphologies,particle sizes,zeta potentials,and pore structure parameters of EDMSNs-NH2 and EDMSNs-PEI were characterized by transmission electron microscopy,dynamic light scattering and nitrogen adsorption-desorption.Both EDMSNs-NH2 and EDMSNs-PEI display obvious dual-mesoporous structure and well-defined spherical morphology.Average dynamic sizes of EDMSNs-NH2 and EDMSNs-PEI are 343.2 and 338.9 nm,while zeta potentials of EDMSNs-NH2 and EDMSNs-PEI are+18 and+43 mV,respectively.The results of gel electrophoresis,CCK-8 assay,and fluorescence microscope show that the pDNA loading amount of EDMSNs-PEI is 8%,much higher than that(1%)of EDMSNs-NH2.In addition,EDMSNs-PEI exhibits lower cytotoxicity than that of pure PEI polymers and lipofectamine2000.When the mass ratio of EDMSNs-PEI/pDNA is 33:1,the transfection rate of the 293T cells by EDMSNs-PEI/pDNA reaches a maximum at 72 h.In conclusion,EDMSNs-PEI,compared with EDMSNs-NH2,has a higher positive potential and pDNA loading capacity,which can be used as a promising non-viral inorganic gene vector for the treatment of diseases such as malignant glioma.

关 键 词:双介孔氧化硅球 聚乙烯亚胺 基因治疗 恶性胶质瘤 

分 类 号:TQ174[化学工程—陶瓷工业]

 

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