DNA誘導(dǎo)金納米粒子自組裝及其生物性能研究
發(fā)布時(shí)間：2019-05-14 19:06
【摘要】：近年來,以金納米粒子、聚合物和生物大分子為組裝基元,用自組裝方法構(gòu)建的金納米粒子生物復(fù)合體系在藥物運(yùn)輸、基因調(diào)控和腫瘤成像及治療等領(lǐng)域取得了廣泛的應(yīng)用。而DNA因其優(yōu)異的生物相容性、可編程性和特殊的堿基識別能力又是構(gòu)建金納米粒子組裝體最有效的手段之一。遺憾的是,目前DNA誘導(dǎo)的金納米粒子組裝體因其在生理鹽水和血清中不穩(wěn)定使其在生物醫(yī)學(xué)領(lǐng)域的應(yīng)用仍然非常有限。本論文以兩種不同尺寸的金納米粒子為組裝基元,利用單鏈DNA的堿基互補(bǔ)作用將其組裝成具有“核-衛(wèi)星”形狀的金納米粒子組裝體。通過改變組裝條件,實(shí)現(xiàn)了“核-衛(wèi)星”金納米粒子組裝體的可控組裝,并對“核-衛(wèi)星”金納米粒子組裝體在生理鹽水和胎牛血清中的穩(wěn)定性、聚乙二醇(PEG)修飾的“核-衛(wèi)星”狀金納米粒子組裝體的細(xì)胞毒性和熒光標(biāo)記特性進(jìn)行了表征。本論文取得的主要研究成果如下:1.分別制備了3.9 nm的“衛(wèi)星”-金納米粒子(Satellite-Au NPs)和18.7 nm的“核”-金納米粒子(Core-Au NPs),并將它們分別與兩條互補(bǔ)的DNA單鏈A'和c A'進(jìn)行偶合,最后實(shí)現(xiàn)“核-衛(wèi)星”狀金納米粒子組裝體(CSNAs)的可控自組裝。研究發(fā)現(xiàn),c A'與Core-Au NPs和Satellite-Au NPs-A'與Core-Au NPs-c A'的比例對組裝體表面組裝的“衛(wèi)星”個(gè)數(shù)起著重要的作用,增加c A'與Core-Au NPs或Satellite-Au NPs-A'與Core-Au NPs-c A'的比例,“核-衛(wèi)星”金納米粒子組裝體的“衛(wèi)星”個(gè)數(shù)將增加。2.在“核-衛(wèi)星”金納米粒子組裝體溶液中分別加入Na Cl溶液和胎牛血清,表征其在Na Cl溶液和血清中的穩(wěn)定性。耐鹽性實(shí)驗(yàn)表明,“核-衛(wèi)星”金納米粒子組裝體在生理鹽水中具有很好的穩(wěn)定性(12 h),而且Core-Au NPs-c A'和Satellite-Au NPs-A'的比例為1:160制備的組裝體可以承受高達(dá)3倍生理鹽水的濃度(3.0%)。血清實(shí)驗(yàn)表明,“核-衛(wèi)星”金納米粒子組裝體在10%的胎牛血清中培養(yǎng)6 h后仍然可以保持完整的“核-衛(wèi)星”的形貌,說明“核-衛(wèi)星”金納米粒子組裝體具有很好的抵抗核酸酶分解的能力。3.進(jìn)行了PEG修飾的“核-衛(wèi)星”金納米粒子組裝體的細(xì)胞毒性和熒光標(biāo)記表征。細(xì)胞毒性實(shí)驗(yàn)表明,修飾了PEG的“核-衛(wèi)星”金納米粒子組裝體在5~50?mol/L的濃度范圍內(nèi)可以維持90%以上的細(xì)胞存活率。熒光實(shí)驗(yàn)表明,修飾了異硫氰酸熒光素的組裝體在488 nm的激發(fā)光源下能夠發(fā)出黃綠色熒光,為熒光檢測提供了可能。
[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.
【學(xué)位授予單位】：華南理工大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2015
【分類號】：TB383.1;O614.123
本文編號：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.
【學(xué)位授予單位】：華南理工大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2015
【分類號】：TB383.1;O614.123
【參考文獻(xiàn)】
相關(guān)期刊論文 前2條
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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