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类型 基础研究 预答辩日期 2017-11-17
开始(开题)日期 2014-12-10 论文结束日期 2017-10-13
地点 金陵院123 论文选题来源 国家自然科学基金项目     论文字数 5 (万字)
题目 SERS活性脂质体复合药物载体的构建及其与肿瘤细胞相互作用研究
主题词 脂质体复合药物载体,金属纳米颗粒,表面增强拉曼散射,肿瘤诊疗
摘要 脂质体复合药物载体可以同时实现药物在肿瘤细胞中的持续递送、载体在细胞内分布的实时监控和协同治疗,这使得脂质体复合药物载体成为肿瘤诊断和治疗一体化研究的新热点。本论文提出了一种SERS活性脂质体-金属复合药物载体,该药物载体可利用表面增强拉曼光谱(SERS)作为脂质体的示踪信号,并将其应用于载体在肿瘤细胞内的药物可控释放及载体与细胞相互作用的研究,为实现基于脂质体-金属复合药物载体的肿瘤诊疗打下了技术基础。 本论文的主要创新性研究成果如下: (1) 设计制备了一种SERS活性的脂质体复合药物载体。首先,以金纳米颗粒为SERS基底,在其表面标记拉曼分子并修饰聚合物,得到功能化的金纳米颗粒;然后,在脂质体表面上吸附功能化金纳米颗粒,构建了脂质体-金属复合药物载体,并对其SERS特性进行研究;最后,将制备的SERS活性脂质体复合药物载体与细胞共培养,并对其细胞内SERS特性进行评估。实验结果表明,用SERS信号可以对复合药物载体在肿瘤细胞内的分布进行监测。 (2) 研究了脂质体-金属复合药物载体与肿瘤细胞间的相互作用。实验中,在银纳米颗粒表面标记拉曼分子并吸附到酸敏感脂质体的表面,构建了新型的复合药物载体;将制备的SERS活性脂质体复合药物载体与肿瘤细胞共培养,并对复合药物载体与肿瘤细胞之间的相互作用进行监测。实验结果表明,该药物载体进入细胞后分布在酸性的溶酶体中并发生药物释放;药物分子扩散出溶酶体,布满整个细胞质。采用SERS光谱有效避免了与药物荧光光谱的重叠,实现了对载体和药物分子在肿瘤细胞内的动态分布的同时示踪,为完善药物载体和细胞间的相互作用与细胞内高效释药提供了实验依据。 (3) 研究了金属纳米颗粒对脂质体的入胞途径和细胞内转运过程的影响。在实验过程中,先借助拉曼标记分子MMTAA作为偶联温度敏感脂质体和金核银壳纳米颗粒的偶联剂,制备了基于脂质体-金核银壳纳米颗粒的复合药物载体;再利用SERS信号对复合药物载体的入胞途径和细胞内转运过程进行监测。实验结果表明,该复合药物载体的细胞行为与空白脂质体类似,主要通过网格蛋白介导的内吞方式进入细胞并滞留在溶酶体中。此外,脂质体也不会影响金核银壳纳米颗粒的SERS特性和光热特性。该研究揭示了脂质体-金属复合药物载体能够进行模块化的设计,有望为肿瘤的诊疗一体化提供一个全新的思路。
英文题目 construction of liposomal nanocarriers and their interactions with tumor cells
英文主题词 liposomal nanohybrids, metal nanoparticles, surface-enhanced Raman scattering, tumor theranostics
英文摘要 The drug delivery systems based on liposome-nanoparticle hybrids enable tumor localization through imaging and simultaneously drug delivery, which makes liposome-nanoparticle hybrids new trends for the investigation of multifunctional drug carriers. Here, the liposome-metal nanohybrid has been proposed to develop a novel liposomal nanocarrier with surface-enhanced Raman scattering (SERS) activity. The SERS signals generated from metal nanoparticles are utilized to track the locations of nanohybrids in tumor cells, which is further employed to study the interactions between liposomal nanocarriers and tumor cells. The experimental results are fundamental for the realization of tumor theranostics based on liposome-metal nanohybrids. The main innovations of this thesis are listed as follows: (1) A liposome-gold nanoparticle hybrids with SERS activity was designed and fabricated. First, functionalized gold nanoparticles were achieved by coating the surfaces of the nanoparticles with Raman reporters and polymers, sequentially. Then, the liposome-gold nanoparticle hybrids were constructed by adsorbing the functionalized gold nanoparticles onto the liposomes, and the SERS activity of the nanohybrids was also studied. Finally, the liposome-gold nanoparticle hybrids were incubated with SKBR3 cells, in which the intracellular SERS performance was evaluated. The experimental results showed that SERS signals could be used to monitor the intracellular locations of nanohybrids. (2) SERS technique was employed to investigate the interactions between the pH-sensitive liposome-silver nanoparticle hybrids and tumor cells. In the experiment, the nanohybrids were achieved by anchoring the Raman reporter molecules tagged silver nanoparticles onto the surfaces of pH-sensitive liposomes. The SERS-active nanohybrids were incubated with HeLa cells, in which the interactions between nanohybrids and HeLa cells were investigated. The experimental results showed that the liposome-silver nanoparticle hybrids were located in the acidic lysosomes, where drug was released. Then the drug molecules diffused across the lysosomes and spread over the whole cytoplasm. The intracellular locations of nanohybrids and drugs were tracked simultaneously using SERS-fluorescence dual modal spectroscopy, which avoided the spectral overlap and provided experimental basis for the carrier-cell interactions and intracellular drug release. (3) The intracellular behaviors of liposome-metal nanohybrids were investigated by SERS technique in order to make insights into the influences of metal nanoparticles. First, MMTAA, a Raman reporter molecule, was employed to conjugate the thermo-sensitive liposomes and gold/silver core-shell nanoparticles. Then, the endocytosis pathway and intracellular fate of the nanohybrids were monitored by SERS signals. Finally, it was revealed that the nanohybrids entered SKBR3 cells through clathrin-mediated endocysotis and remained in lysosomes, which was quite consistent with that of pure liposomes. In addition, liposomes would not affect the SERS activity and photothermal effect of metal nanoparticles, neither. The experimental results suggested a universal rule in the modular design of multifunctional nanohybrids, which was expected to provide a new route for tumor theranostics.
学术讨论
主办单位时间地点报告人报告主题
先进光子学中心 Jan 3,2017 金陵院123 朱丹 Liposomal Texaphyrin Theranostics for Metastatic Liver Cancer
先进光子学中心 Jun 27,2016 金陵院123 朱丹 Magnetically Guided Protein Transduction by Hybrid Nanogel Chaperones with Iron Oxide Nanoparticles
先进光子学中心 Mar 21,2016 金陵院123 朱丹 A Telomerase-Specific Doxorubicin-Releasing Molecular Beacon for Cancer Theranostics
先进光子学中心 Oct 28,2015 金陵院123 朱丹 Intracellular Delivery of Functional Proteins and Native Drugs by Cell-Penetrating Poly(disulfide)s
先进光子学中心 Apr 11, 2017 金陵院123 朱丹 Controlling Spatial Heat and Light Distribution by Using Photothermal Enhancing Auto-Regulated Liposomes (PEARLs)
先进光子学中心 Jul 9,2015 金陵院123 朱丹 In Vivo Remote Control of Reactions in Caenorhabditis elegans by Using Supermolecular Nanohybrids of Carbon Nanotubes and Liposomes
先进光子学中心 Nov 5,2014 金陵院123 朱丹 Super-Resolution SERS Imaging beyond the Single-Molecule Limit: An Isotope-Edited Approach
先进光子学中心 May 13,2014 金陵院123 朱丹 SERS-Encoded Nanogapped Plasmonic Nanoparticles: Growth of Metallic Nanoshell by Templating Redox-Active Polymer Brushes
     
学术会议
会议名称时间地点本人报告本人报告题目
JSAP-OSA Joint Symposia 2013 Sep 16-20, 2013 Kyoto, Japan Liposome-silver nanoparticles hybrid as a SERS traceable drug carrier
The 3rd International Symposium Laser Interaction with Matter (LIMIS 2014) Nov 2-5, 2014 Nanjing, China Gold nanoparticles decorated liposomes and their SERS performance in tumor cells.
2016 International Conference on Frontiers of Composite Materials (ICFCM2016) Nov 19-21, 2016 Auchland, New Zealand Metal-liposome nanocomposites with SERS activity for photothermal release in cancer cells.
The 6th International Conference on Metamaterials, Photonics Crystals and Plasm Aug 4-7, 2015 New York, USA Uptake of Liposomal Hybrids in Tumor Cells Using Surface-Enhanced Raman Spectroscopy
     
代表作
论文名称
Ag@4ATP-coated liposomes: SERS traceable delivery vehicles for living cells
Wavenumber-Intensity Joint SERS Encoding Using Silver Nanoparticles for Tumor Cell Targeting
 
答辩委员会组成信息
姓名职称导师类别工作单位是否主席备注
倪晓武 正高 教授 博导 南京理工大学
王亚伟 正高 教授 博导 江苏大学
张雄 正高 教授 博导 东南大学
恽斌峰 正高 教授 博导 东南大学
顾兵 正高 教授 博导 东南大学
      
答辩秘书信息
姓名职称工作单位备注
宗慎飞 副高 副研究员 东南大学