国家重大科学研究计划项目“生物医学纳米材料对血细胞作用的研究”工作进展与讨论

结合多模态纳米探针进行淋巴癌等血液恶性肿瘤的在体分子成像与疗效评价研究

东南大学 顾宁 教授


1. 纳米颗粒的吸附对有衬底的磷脂分子层性质影响的模拟研究

有衬底的磷脂分子层结构稳定,可利用表面研究技术加以研究,从而获得分子层面上的作用信息。研究纳米颗粒与有衬底的磷脂分子层的作用,对纳米颗粒与细胞膜作用的研究、纳米颗粒-有衬底的磷脂分子层构成的纳米器件研究具有重要的启发作用。

我们用分子动力学的方法从分子水平上研究了纳米颗粒加热、表面电荷等对DPPC双分子层性质的影响。结果表明:1) 纳米颗粒在DPPC分子层上的吸附行为主要由颗粒的表面电荷调控,颗粒的加热对之影响不大;2)加热的颗粒可致使与之接近的DPPC分子的热运动增加,界面水和块体水的动力学差异致使靠近衬底的DPPC单层比远离衬底的DPPC单层具有更强的热运动特性和更弱的扩散能力;3)吸附在DPPC分子层上纳米颗粒附近的DPPC分子的性质(扩散特性、序参数等)主要由纳米颗粒的表面电荷性质调控。
Xubo Lin, Changling Wang, Meng Wang, Kun Fang, Ning Gu. Computer Simulation of the Effects of Nanoparticles' Adsorption on the Properties of Supported Lipid Bilayer. Journal of Physical Chemistry C 2012, 116(33), 17960-17968.


Fig.1 Temperature distribution along the z-axis for all simulation systems.

2. 纳米银诱导不同代数Ba/F3-BCR-ABL细胞效应的比较研究

利用细胞系进行纳米材料体外生物效应的评价是纳米材料生物效应研究的一种重要手段。然而越来越多的研究认为,一些细胞系的过度传代培养将影响其细胞特性。基于此,我们将融合基因BCR-ABL细胞转染永生化小鼠前B细胞Ba/F3,使其不再依赖于IF-3的生长。然后进行连续传代培养研究其稳定性以及其对纳米银的细胞反应性。结果显示,短期传代不会影响Ba/F3-BCR-ABL细胞的稳定性,纳米银诱导不同代数细胞效应没有显著差异。因此,使用这样培养的细胞进行细胞效应评价时可以提供一致结果。
GUO DaWei, ZHANG XiuYan, HUANG ZhiHai, ZHOU XueFeng, ZHU LingYing, ZHAO Yun, GU Ning, Comparison of cellular responses across multiple passage numbers in Ba/F3-BCR-ABL cells induced by silver nanoparticles, SCIENCE CHINA (Life Sciences) 2012, 55(10), 898-905.


Fig.2 Characterization of silver nanoparticles. A and B, The morphology and size distribution of the silver nanoparticles were characterized by transmission electron microscopy (TEM). The mean diameter was 29.7 nm. C, The AgNPs were characterized by the UV-Vis absorption spectrum, and the maximum absorption was 417 nm. D and E, Particles size (diameter) and surface charge in an aqueous solution were measured by dynamic light scattering (DLS) and were (71.83±0.74) 23.0nm and (±1.01) mV, respectively.


Fig. 3 Uptake and localization of AgNPs in Ba/F3-BCR-ABL cells at multiple passage numbers. A, TEM images of ultrathin sections of Ba/F3-BCR-ABL cells treated with AgNPs (12 μg 1) for 24 h showed that the particles weremL internalized and could be sequestered in vacuoles, which were visible in the cells as black, and the electron-dense spots are indicated by arrows. B, The cells treated with AgNPs (12 μ 1) for 24 h were digested in nitric acid, and the silver concentration wasg mL determined by ICP-MS. “+” denotes that cells were treated with AgNPs.


Fig. 4 Effects of AgNP exposure on Ba/F3-BCR-ABL cell viability at multiple passage numbers. After a 24-h exposure of AgNPs at multiple concentrations, cell viability was determined with a Cell Counting Kit-8 (CCK-8). The data were a summary of three independent experiments and expressed as the means±SD. * denotes a significant reduction in cell viability (P<0.05) compared with the control.


Fig. 5 Effect of passage number and AgNP exposure on apoptosis of Ba/F3-BCR-ABL cells. A and B, The representative images and summaries of apoptosis of the AgNP-treated (10 μ g 1) and untreated Ba/F3-BCR-ABLmL cells at multiple passage numbers are presented. C and D, The representative images and summaries of apoptosis of the 1st passage Ba/F3-BCR-ABL cells induced by AgNP exposure at various concentrations are presented.


Fig. 6 Effect of passage number and AgNP exposure on the cell cycle of Ba/F3-BCR-ABL cells. A and B, The representative images and summaries of the cell cycle of the AgNP-treated (10 μ g 1) and untreated Ba/F3-BCR-ABLmL cells at multiple passage numbers are presented. C and D, The representative images and summaries of the cell cycle of the 1st passage Ba/F3-BCR-ABL cells induced by AgNP exposure at various concentrations are presented.

3. 应用于动脉粥样硬化的CT-荧光双模态碘油纳米乳研究进展

巨噬细胞在动脉粥样硬化的发生发展过程中起着关键的作用,已被作为诊断动脉粥样硬化的重要标记物。多模态造影剂是指通过一次注射一种造影剂,可同时用于不同影像设备如CT、MRI、荧光等的检测,实现多种显像模式的优势互补。本文主要介绍一种CT-荧光双模态碘油纳米乳造影剂,该造影剂将CT的高分辨率和荧光的高灵敏度结合在一起,实现了两种造影模式的优势互补,不仅可对病变区域进行CT造影,又可荧光检测,有利于得到更全面更精准的诊断信息。CT-荧光双模态碘油纳米乳,选择碘化油注射液作为CT造影部分,油溶性CdSe/ZnS量子点作为光学探针部分,借助于搅拌和超声乳化的方法形成稳定的水包油纳米乳,得到尺寸均一(80nm)、稳定性好、生物相容性高的透明清亮的双模态碘油纳米乳,并测定其CT和荧光效果。其后将碘油纳米乳与细胞进行共培养,在细胞水平上进行micro CT和共聚焦荧光效果的验证。该碘油纳米乳造影剂主要针对动脉粥样硬化的造影,由于斑块中含有大量的巨噬细胞,纳米乳可通过巨噬细胞的吞噬作用进入斑块,因此选择了腹主动脉有动脉粥样硬化斑块的新西兰大白兔作为模型,在动物水平上进行CT造影,之后对离体的血管进行荧光成像。结果显示,该碘油纳米乳可通过巨噬细胞的被动靶向作用,实现对粥样硬化模型的CT/荧光双模态造影,在医学诊断中具有发展潜力。
Jiali Ding, Yuehua Wang (Co-first author), Ming Ma, Yu Zhang, Shanshan Lu, Yanni Jiang, Chunmei Qi, Shouhua Luo, Ge Dong, Song Wen, Yanli An, Ning Gu. , CT/fluorescence dual-modal nanoemulsion platform for investigating atherosclerotic plaques, Biomaterials 2013, 34, 209-216.

4. H2O2响应的超声磁共振双模式造影剂的研究

外界环境,如疾病或外界物质的引入会引起人体内活性氧(ROS)的产生,H2O2是ROS的一种,它能自由扩散并具有相对长的稳定时间,因此,近年来H2O2成为许多疾病发生、发展过程的诊断标记物。本实验基于发现普鲁士蓝纳米颗粒(PBNPs)在中性环境条件下(pH=7.4)具有类过氧化氢酶的作用催化H2O2产生氧气的原理,研究了产生的O2超过饱和浓度后能形成O2自由气泡,验证了PBNPs在体外超声和磁共振双模式显影以及在炎症动物模型状态下的体内超声和磁共振双模式显影。
Fang Yang, Sunling Hu, Yu Zhang, Xiaowei Cai, Yan Huang, Feng Wang, Song Wen, Gaojun Teng, Ning Gu. A Hydrogen Peroxide-Responsive O2 Nanogenerator for Ultrasound and Magnetic-Resonance Dual Modality Imaging. Adv. Mater. 20112, 24(38): 5205–5211