DNA诱导金纳米粒子自组装及其生物性能研究
发布时间：2019-05-14 19:06
【摘要】：近年来,以金纳米粒子、聚合物和生物大分子为组装基元,用自组装方法构建的金纳米粒子生物复合体系在药物运输、基因调控和肿瘤成像及治疗等领域取得了广泛的应用。而DNA因其优异的生物相容性、可编程性和特殊的碱基识别能力又是构建金纳米粒子组装体最有效的手段之一。遗憾的是,目前DNA诱导的金纳米粒子组装体因其在生理盐水和血清中不稳定使其在生物医学领域的应用仍然非常有限。本论文以两种不同尺寸的金纳米粒子为组装基元,利用单链DNA的碱基互补作用将其组装成具有“核-卫星”形状的金纳米粒子组装体。通过改变组装条件,实现了“核-卫星”金纳米粒子组装体的可控组装,并对“核-卫星”金纳米粒子组装体在生理盐水和胎牛血清中的稳定性、聚乙二醇(PEG)修饰的“核-卫星”状金纳米粒子组装体的细胞毒性和荧光标记特性进行了表征。本论文取得的主要研究成果如下:1.分别制备了3.9 nm的“卫星”-金纳米粒子(Satellite-Au NPs)和18.7 nm的“核”-金纳米粒子(Core-Au NPs),并将它们分别与两条互补的DNA单链A'和c A'进行偶合,最后实现“核-卫星”状金纳米粒子组装体(CSNAs)的可控自组装。研究发现,c A'与Core-Au NPs和Satellite-Au NPs-A'与Core-Au NPs-c A'的比例对组装体表面组装的“卫星”个数起着重要的作用,增加c A'与Core-Au NPs或Satellite-Au NPs-A'与Core-Au NPs-c A'的比例,“核-卫星”金纳米粒子组装体的“卫星”个数将增加。2.在“核-卫星”金纳米粒子组装体溶液中分别加入Na Cl溶液和胎牛血清,表征其在Na Cl溶液和血清中的稳定性。耐盐性实验表明,“核-卫星”金纳米粒子组装体在生理盐水中具有很好的稳定性(12 h),而且Core-Au NPs-c A'和Satellite-Au NPs-A'的比例为1:160制备的组装体可以承受高达3倍生理盐水的浓度(3.0%)。血清实验表明,“核-卫星”金纳米粒子组装体在10%的胎牛血清中培养6 h后仍然可以保持完整的“核-卫星”的形貌,说明“核-卫星”金纳米粒子组装体具有很好的抵抗核酸酶分解的能力。3.进行了PEG修饰的“核-卫星”金纳米粒子组装体的细胞毒性和荧光标记表征。细胞毒性实验表明,修饰了PEG的“核-卫星”金纳米粒子组装体在5~50?mol/L的浓度范围内可以维持90%以上的细胞存活率。荧光实验表明,修饰了异硫氰酸荧光素的组装体在488 nm的激发光源下能够发出黄绿色荧光,为荧光检测提供了可能。
[Abstract]:In recent years, the gold nanoparticles biocomposite system constructed by self-assembly method with gold nanoparticles, polymers and biological macromolecules as assembly elements has been widely used in drug transportation, gene regulation, tumor imaging and treatment. Because of its excellent biocompatibility, programmable property and special base recognition ability, DNA is one of the most effective means to construct gold nanoparticles assembly. Unfortunately, the application of DNA-induced gold nanoparticles in biomedicine is still very limited because of its instability in saline and serum. In this paper, two kinds of gold nanoparticles of different sizes are used as assembly elements, and the gold nanoparticles with "nucleus-satellite" shape are assembled by the base complementarity of single chain DNA. The controllable assembly of "nuclear-satellite" gold nanoparticles assembly was realized by changing the assembly conditions, and the stability of "nuclear-satellite" gold nanoparticles assembly in saline and fetal bovine serum was also realized. The cytotoxicity and fluorescence labeling characteristics of "nucleus-satellite" gold nanoparticles modified by polyethylene glycol (PEG) were characterized. The main research results of this paper are as follows: 1. 3.9 nm "satellite" gold nanoparticles (Satellite-Au NPs) and 18.7 nm "core" gold nanoparticles (Core-Au NPs),) were prepared and coupled with two complementary DNA single chain A' and c A', respectively. Finally, the controllable self-assembly of "core-satellite" gold nanoparticles assembly (CSNAs) is realized. It is found that the ratio of c A'to Core-Au NPs and Satellite-Au NPs-A' to Core-Au NPs-c A 'plays an important role in the number of "satellites" assembled on the surface of the assembly. By increasing the ratio of c A'to Core-Au NPs or Satellite-Au NPs-A' to Core-Au NPs-c A', the number of "satellites" in the "nuclear-satellite" gold nanoparticles assembly will increase. 2. Na Cl solution and fetal bovine serum were added to the "nuclear-satellite" gold nanoparticles assembly solution, respectively, and their stability in Na Cl solution and serum was characterized. The salt tolerance test shows that the "core-satellite" gold nanoparticles assembly has good stability in saline (12 h),). Moreover, the ratio of Core-Au NPs-c A'to Satellite-Au NPs-A' is 1 脳 160, and the assembly can withstand up to 3 times the concentration of saline (3.0%). Serum experiments showed that the "nuclear-satellite" gold nanoparticles could still maintain a complete "nuclear-satellite" morphology after being cultured in 10% fetal bovine serum for 6 hours. It is shown that the "nucleus-satellite" gold nanoparticles assembly has a good ability to resist nuclease decomposition. The cytotoxicity and fluorescence labeling of PEG modified "nucleus-satellite" gold nanoparticles were characterized. Cytotoxicity tests showed that the "nuclear-satellite" gold nanoparticles modified with PEG could maintain more than 90% cell survival in the concentration range of 5~50?mol/L. Fluorescence experiments show that the assembly modified with fluorescein isothiocyanate can emit yellow-green fluorescence under the excitation light source of 488 nm, which makes it possible for fluorescence detection.
【学位授予单位】：华南理工大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：TB383.1;O614.123
本文编号：2476974
[Abstract]:In recent years, the gold nanoparticles biocomposite system constructed by self-assembly method with gold nanoparticles, polymers and biological macromolecules as assembly elements has been widely used in drug transportation, gene regulation, tumor imaging and treatment. Because of its excellent biocompatibility, programmable property and special base recognition ability, DNA is one of the most effective means to construct gold nanoparticles assembly. Unfortunately, the application of DNA-induced gold nanoparticles in biomedicine is still very limited because of its instability in saline and serum. In this paper, two kinds of gold nanoparticles of different sizes are used as assembly elements, and the gold nanoparticles with "nucleus-satellite" shape are assembled by the base complementarity of single chain DNA. The controllable assembly of "nuclear-satellite" gold nanoparticles assembly was realized by changing the assembly conditions, and the stability of "nuclear-satellite" gold nanoparticles assembly in saline and fetal bovine serum was also realized. The cytotoxicity and fluorescence labeling characteristics of "nucleus-satellite" gold nanoparticles modified by polyethylene glycol (PEG) were characterized. The main research results of this paper are as follows: 1. 3.9 nm "satellite" gold nanoparticles (Satellite-Au NPs) and 18.7 nm "core" gold nanoparticles (Core-Au NPs),) were prepared and coupled with two complementary DNA single chain A' and c A', respectively. Finally, the controllable self-assembly of "core-satellite" gold nanoparticles assembly (CSNAs) is realized. It is found that the ratio of c A'to Core-Au NPs and Satellite-Au NPs-A' to Core-Au NPs-c A 'plays an important role in the number of "satellites" assembled on the surface of the assembly. By increasing the ratio of c A'to Core-Au NPs or Satellite-Au NPs-A' to Core-Au NPs-c A', the number of "satellites" in the "nuclear-satellite" gold nanoparticles assembly will increase. 2. Na Cl solution and fetal bovine serum were added to the "nuclear-satellite" gold nanoparticles assembly solution, respectively, and their stability in Na Cl solution and serum was characterized. The salt tolerance test shows that the "core-satellite" gold nanoparticles assembly has good stability in saline (12 h),). Moreover, the ratio of Core-Au NPs-c A'to Satellite-Au NPs-A' is 1 脳 160, and the assembly can withstand up to 3 times the concentration of saline (3.0%). Serum experiments showed that the "nuclear-satellite" gold nanoparticles could still maintain a complete "nuclear-satellite" morphology after being cultured in 10% fetal bovine serum for 6 hours. It is shown that the "nucleus-satellite" gold nanoparticles assembly has a good ability to resist nuclease decomposition. The cytotoxicity and fluorescence labeling of PEG modified "nucleus-satellite" gold nanoparticles were characterized. Cytotoxicity tests showed that the "nuclear-satellite" gold nanoparticles modified with PEG could maintain more than 90% cell survival in the concentration range of 5~50?mol/L. Fluorescence experiments show that the assembly modified with fluorescein isothiocyanate can emit yellow-green fluorescence under the excitation light source of 488 nm, which makes it possible for fluorescence detection.
【学位授予单位】：华南理工大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：TB383.1;O614.123
【参考文献】
相关期刊论文 前2条
1 ZHOU WeiYa;ZHANG XiaoXian;ZHAO Duan;GAO Min;XIE SiShen;;ZnO nanorods:morphology control, optical properties, and nanodevice applications[J];Science China(Physics,Mechanics & Astronomy);2013年12期
2 Yafei Zhang;Li Franklin Duan;Yaozhong Zhang;Jian Wang;Huijuan Geng;Qing Zhang;;Advances in Conceptual Electronic Nanodevices based on 0D and 1D Nanomaterials[J];Nano-Micro Letters;2014年01期
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