Liu Zhen-bao, Zhou Bo, Wang Hai-yan, Zhang Hai-ling, Liu Lan-xia, Zhu Dun-wan, Leng Xi-gang
Tianjin Key Laboratory of Biomedical Materials, Institute of Biomedical Engineering, CAMS and PUMC, Tianjin 300192, China.
Zhongguo Yi Xue Ke Xue Yuan Xue Bao. 2010 Aug;32(4):449-55. doi: 10.3881/j.issn.1000-503X.2010.04.020.
To investigate the cytotoxic effect of multi-walled carbon nanotubes (MWCNTs) on human liver L02 cells and its relevant mechanism.
MWCNTs, carboxyl modification MWCNTs (MWCNTs-COOH), and hydroxyl modification 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 200 μg/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.
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. X-ray 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 > MWCNTs-COOH > MWCNTs-OH.
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.
研究多壁碳纳米管(MWCNTs)对人肝L02细胞的细胞毒性作用及其相关机制。
采用透射电子显微镜、扫描电子显微镜和X射线光电子能谱对MWCNTs、羧基修饰的MWCNTs(MWCNTs-COOH)和羟基修饰的MWCNTs(MWCNTs-OH)进行表征。将浓度为12.5、25、50、100和200μg/ml的碳纳米管分别与人肝L02细胞孵育24、48和72小时。通过水溶性四氮唑盐法评估细胞活力,采用2',7'-二氯二氢荧光素二乙酸酯法检测碳纳米管诱导的细胞内活性氧。
透射电子显微镜显示三种MWCNTs的平均外径(10至20nm)和平均长度(10至30μm)相似。扫描电子显微镜表明三种MWCNTs具有相似的表面形貌。X射线光电子能谱表明,MWCNTs-COOH和MWCNTs-OH分别在289和286ev处具有相对较高的峰面积,表明它们分别被羧基和羟基修饰。水溶性四氮唑盐法显示,与MWCNTs相比,MWCNTs-COOH的细胞毒性较小,而MWCNTs的细胞毒性略高于MWCNTs-OH。诱导细胞内活性氧增加的能力顺序为:MWCNTs>MWCNTs-COOH>MWCNTs-OH。
羧基和羟基修饰MWCNTs在一定程度上提高了MWCNTs的生物相容性。低浓度时MWCNTs-COOH比MWCNTs具有更好的相容性,48小时后MWCNTs-OH比MWCNTs表现出更好的相容性。细胞与具有不同化学表面的MWCNTs之间的相互作用可能涉及不同的机制。