功能化多壁碳纳米管对L02细胞的作用  

Effect of Functionalized Multi-walled Carbon Nanotubes on L02 Cells

作　　者：刘珍宝[1] 周波[1] 王海燕[1] 张海玲[1] 刘兰霞[1] 朱敦皖[1] 冷希岗[1] 

机构地区：[1]中国医学科学院北京协和医学院生物医学工程研究所天津市生物材料重点实验室,天津300192

出　　处：《中国医学科学院学报》2010年第4期449-455,共7页Acta Academiae Medicinae Sinicae

基　　金：科技部科研院所社会公益研究专项基金(2005DIB1J094);国家重大研究计划项目(2006CB933203)~~

摘　　要：目的探讨羧基化及羟基化多壁碳纳米管对肝细胞的毒性及相关机制。方法利用透射电镜、扫描电镜、X-射线光电子能谱仪对原始多壁碳纳米管、羧基化多壁碳纳米管和羟基化多壁碳纳米管进行表征。将浓度分别为12.5、25、50、100、200μg/ml的3种碳纳米管分别与人正常肝细胞系L02细胞共育24、48、72h。采用水溶性四氮唑法进行细胞毒性评价,用2',7'-二氯荧光黄双乙酸盐法检测细胞内活性氧(ROS)的生成。结果透射电镜显示3种碳纳米管平均管径均为10～20nm,长度均为10～30μm;扫描电镜显示碳纳米管形貌相似;X-射线光电子能谱仪分析显示功能化多壁碳纳米管分别在289和286ev处特征峰峰值明显增高。水溶性四氮唑法检测显示碳纳米管的细胞毒性作用呈现浓度依赖性并与作用时间具有一定关系。羧基化多壁碳纳米管比原始多壁碳纳米管及羟基化多壁碳纳米管的细胞毒性小。诱导细胞内ROS升高的次序为原始多壁碳纳米管>羧基化多壁碳纳米管>羟基化多壁碳纳米管;碳纳米管诱导细胞内活性氧含量与共育时间有关:碳纳米管与细胞作用36h以内时,细胞内ROS含量随时间逐渐增加,36h后细胞内ROS含量随作用时间延长逐渐降低。结论羧基化多壁碳纳米管比原始多壁碳纳米管在低浓度时具有更好的生物相容性,羟基化多壁碳纳米管比原始多壁碳纳米管在48h后表现出稍好的生物相容性。不同化学表面性质的碳纳米管对细胞活力的影响不同,其与细胞的相互作用可能存在不同机制。Objective To investigate the cytotoxic effect of multi-walled carbon nanotubes(MWCNTs)on human liver L02 cells and its relevant mechanism.Methods MWCNTs,carboxyl modificated MWCNTs(MWCNTs-COOH),and hydroxyl modificated MWCNTs(MWCNTs-OH)were characterized by transmission electron microscopy,scanning electron microscopy,and X-ray photoelectron spectroscopy.The carbon nanotubes at concentrations of 12.5,25,50,100,and 200μg /ml were incubated with human liver L02 cells for 24,48 and 72 hours,respectively.The cell viability was evaluated by water soluble tetrazolium salts assay and the intercellular reactive oxygen species induced by the carbon nanotubes were detected by 2’,7’-dichlorodihydrofluorescein diacetate method.Results Transmission electron microscope showed that the average outside diameters(10 to 20 nm)and the average length(10 to 30 μm)of the three MWCNTs were similar.Scanning electron microscope indicated that the three MWCNTs had a similar surface topography.Xray photoelectron spectroscopy demonstrated that the MWCNTs-COOH and MWCNTs-OH had relatively high peak areas at 289 and 286ev,respectively,indicating that they have been modified by carboxyl and hydroxyl groups,respectively.Water soluble tetrazolium salts assay showed that the MWCNTs-COOH was less cytotoxic when compared to MWCNTs which demonstrated to be slightly more cytotoxic than MWCNTs-OH.The capability to induce increase in intracellular reactive oxygen species was in the following order:MWCNTs > MWCNTsCOOH > MWCNTs-OH.Conclusions Modification of MWCNTs with carboxyl group and hydroxyl group improves the biocompatibility of MWCNTs to some extents.MWCNTs-COOH has better compatibility than MWCNTs at the low concentration,and MWCNTs-OH showed better compatibility than MWCNTs after 48 hours.Different mechanisms may be involved in the interaction between cells and the MWCNTs with different chemical surfaces.

关 键 词：多壁碳纳米管 化学修饰 细胞毒性 

分 类 号：R318.08[医药卫生—生物医学工程]