功能化硅壳荧光纳米粒的细胞吞噬研究  

CELLULAR UPTAKE OF FUNCTIONALIZED SILICA-COATED FLUORESCENT NANOPARTICLES

作　　者：赵焕英[1] 王楠[2] 于景娴[3] 杨慧[1] 邓云龙[3] 彭玉龙[3] 

机构地区：[1]首都医科大学神经科学研究所,北京100069 [2]中国人民武装警察部队总医院,北京100039 [3]首都医科大学化学生物学与药学院,北京100069

出　　处：《高分子学报》2009年第7期667-673,共7页Acta Polymerica Sinica

基　　金：国家重点基础研究发展计划(973计划;项目号2006CB500706);国家自然科学基金项目(基金号30670655);北京市属高等学校人才强教深化计划项目

摘　　要：合成了氨基聚酰胺-胺(PAMAM(G1.0))和酯基(PAMAM(G1.5))功能化的两种硅壳荧光纳米粒,通过透射电镜(TEM)、纳米粒度及动电位测定仪(zeta电势)、傅立叶红外光谱仪(FTIR)和热失重分析仪(TGA)进行表征;通过透射电镜(TEM)、共聚焦显微镜(CLSM)、细胞计数试剂盒(CCK-8)实验、流式细胞计数法评价两种硅壳荧光纳米粒进入9L细胞能力的大小、在细胞内的分布情况以及细胞毒性.TEM分析表明,修饰的硅壳纳米粒大小约为60 nm左右,pH=7.4,氨基功能化的纳米粒zeta电势为+19.08,酯基功能化的为-9.01;FTIR和TGA实验进一步证明两种纳米粒被氨基和酯基的功能化.TEM和CLSM结果表明纳米粒主要存在细胞浆中,且能被溶酶体吞噬.CCK-8结果显示两种纳米粒的浓度高达1 mg/mL时仍无明显的毒性作用,且有促细胞增殖作用.流式细胞计数结果表明,细胞摄取纳米粒呈浓度和时间依赖性,氨基比酯基修饰的纳米粒更易进入细胞.Silica-coated fluorescent nanoparticles functionalized with amine-or ester-poly（amido-amide）（PAMAM） were synthesized and characterized by TEM, FTIR, TGA and zeta potential measurements. The efficiency of cellular uptake of nanoparticles, the cytotoxicity of nanoparticles toward 9L cell, and the distribution of nanoparticles in 9L cell were investigated by transmission electron microscopy （TEM）, confocal laser scanning microscopy （ CLSM）, CCK-8 assay and flow cytometry analysis. TEM revealed the average size of nanoparticles was about 60 nm and the zeta potential of nanoparticles modified with amine groups was ＋ 19.08, with ester groups was negative-9.01 at physiological pH , respectively. FTIR and TGA results also testified that the two kind of silica-coated fluorescent nanoparticles were respectively functionalized by amine- and ester- PAMAM. CI_SM demonstrated that most nanoparticles were distributed in cytoplasm and were entrapped in endosomes. CCK-8 analysis showed that both nanoparticles had no apparent toxicity. Flow cytometry analysis indicated that the cellular uptake of nanoparticles by 9L cells was a concentration- and time- dependent endocytic process, and the uptake of amine functionalized nanoparticles was more efficient than that of ester functionalized nanoparticles.

关 键 词：二氧化硅 PAMAM树状分子 细胞吞噬 细胞毒性 

分 类 号：R318.08[医药卫生—生物医学工程]