无机纳米材料的制备及其在生物分析中的应用

发布时间：2018-05-31 12:31

本文选题：量子点 + 荧光共振能量转移　；参考：《东北大学》2015年博士论文

【摘要】：无机纳米材料因其维度的逐渐降低,在光、电、磁、声等方面表现出不同于常规材料的优良的物理和化学性质。无机纳米材料与生物和化学分析技术的深入结合已成为国内外生物医学分析和环境分析领域的前沿和热点问题。然而,目前大部分基于纳米材料的传感器由于受到高消耗、抗干扰能力差以及不稳定等分析性能的限制,仍然处于实验室研究阶段。同时以纳米材料负载的药物的可控性释放亟待研究。基于上述背景,本论文旨在将纳米材料引入到化学／生物传感器以及药物负载和可控性药物释放之中：(1)结合无机纳米材料优良的光学性能,有目的性的设计纳米探针,构建具有较好抗干扰能力、稳定性好以及灵敏度高的分析方法,应用于生物和环境样品的分析；(2)设计合成多功能的纳米复合材料,以实现药物的负载和可控性释放,提高杀菌治疗效率,减少或避免广谱抗菌药物的使用以及物理杀菌带来的组织损伤或水体中营养物质损失。论文的第一章简要介绍了无机纳米材料的种类、物理化学性质和应用。详细介绍了半导体纳米晶和金纳米棒两种无机纳米材料的组成、分类、光学性质以及在生物和环境分析中的应用。论文的第二章合成了以N-乙酰-L-半胱氨酸为稳定剂的量子点(NAC-QDs)和对汞有特异性响应的罗丹明6G衍生物(R6G-D),并对它们进行了红外光谱、核磁共振氢谱表征。基于NAC-QDs和R6G-D对汞离子的荧光响应及它们吸收/发射光谱的重叠,建立了NAC-QDs/R6G-D荧光共振能量转移体系,用于汞的比率荧光检测,并对标准样品和环境水样中的汞离子进行了定量检测。检测汞的线性范围为5-250 μgmL-1,精密度为3.2%(175μg L-1,n=11),检出限为0.75μg L-1(3σ/s,n=11)。该FRET传感体系能够在0-250 μg mL-1浓度范围内随着汞离子的浓度呈现色差变化,可以对汞离子进行半定量可视化检测,辨别性能可达50μg L-1。本文所建立的比率荧光FRET检测平台能够通过双波长荧光强度比成功的避免检测器带来的干扰,具有较高的灵敏度、良好的选择性、且检测过程简单、检测结果准确。本方法能够在降低成本的情况下实现环境及生物样品中待测物的准确定量检测和可视化半定量检测。论文的第三章合成了金纳米棒包覆／卡那霉素负载的中空二氧化硅纳米囊(HSKAurod)。纳米复合材料HSKAurod纳米囊同时具备了金纳米棒的物理光热杀菌性能和杀菌药物卡那霉素的化学杀菌性能,能够在光热治疗的同时实现药物的负载与可控性释放。以中空二氧化硅纳米囊作为药物载体负载广谱抗菌药卡那霉素,将金纳米棒作为近红外光热响应材料包覆于纳米囊表面,避免负载的卡那霉素的泄漏。选取大肠杆菌E. coli BL21为目标菌株,研究了HSKAurod纳米复合材料的杀菌性能和杀菌机理。HSKAurod纳米囊具有很高的光热转换效率,10mgmL-1的材料能在近红外光(785 nm)照射20 min内使环境温度由室温的21℃迅速升至50℃。HSKAurod纳米囊的杀菌性能显著提高,10 mg mL-1的HSKAurod纳米囊在近红外光照射处理20 min后,细菌杀死率几乎达到100%。这表明物理杀菌与化学杀菌的结合可以产生杀菌的协同作用,有效降低了抗菌药物的用量,缩短了光热治疗的时间,降低了物理杀菌带来的生物组织损伤。HSKAurod纳米囊有潜力代替传统的杀菌剂,应用于临床杀菌的辅助治疗。论文的第四章设计并合成了双金属核-壳-壳金-银-金纳米棒(Au-Ag-Au nanorods)作为近红外光区内的光热杀菌材料。Au-Ag-Au nanorods外层金壳能够在近红外光照射下消融而使中间的银壳层暴露出来,实现抗菌的银壳层/Ag+的可控性释放。这种将外层金壳的光热杀菌与中间银壳层/Ag+的可控性释放相结合,赋予了Au-Ag-Au nanorods卓越的杀菌性能。以大肠杆菌E. coli 0157:H7为细菌模板,研究了双金属核-壳-壳纳米棒的杀菌性能和杀菌机理。在低功率的近红外激光照射下,外壳金层使pAu-Ag-Au nanorods(44℃)展现了优于金-银纳米棒(Au-Ag nanorods,39℃)的光热转换性能。与此同时双金属纳米棒的核-壳-壳结构使其具有比Au-Ag nanorods更好的化学稳定性,16天内Au-Ag-Au nanorods的吸收光谱稳定,吸光度无变化。10μgmL-1的Au-Ag-Au nanorods在近红外光照射10 min就能使周围溶液的温度达到44℃,在近红外光照射20 min时对大肠杆菌E. coli 0157:H7的杀死率达到100%。双金属核-壳-壳Au-Ag-Au nanorods用于杀菌,成功避免了广谱抗菌药物的使用,减少了材料的用量,提高了杀菌效率,同时低功率近红外光的使用降低了对生物组织带来的损伤。上述特性使双金属核-壳-壳Au-Ag-Au nanorods有望成为一种新型的纳米光热转换材料用于体内的生物医学研究。论文的第五章首先通过一步法合成了带正电的磁性rGO-Fe3O4-PEI纳米材料,并在还原氧化石墨烯片层上负载大量的双金属核-壳-壳Au-Ag-Au nanorods,得到的rGO-Fe3O4-Au-Ag-Au纳米复合材料作为近红外光热材料,用于细菌的捕获、分离和杀死。以大肠杆菌E. coli O157:H7为目标菌株,分别研究了纳米复合材料对细菌的捕获能力、磁分离能力、光热转换效率以及杀菌性能。rGO-Fe3O4磁性材料的组装,使得rGO-Fe3O4-Au-Ag-Au纳米复合材料在近红外光热杀菌的同时,还能对细菌进行识别捕获并从水体样品中磁分离出来O30 μgmL-1的rGO-Fe3O4-Au-Ag-Au在磁场作用下10 mmin内就能100%识别捕获浓度为1 × 108 cfu mL-1的E. coli O157:H7并将其从溶液中磁分离除去。rGO-Fe3O4自身具有一定的光热转换性能,加上石墨烯材料超好的导热性能,组装后有效提高了纳米复合材料的光热转换效率,25 μg mL-1的rGO-Fe3O4-Au-Ag-Au在近红外光照射10 min可使周围溶液的温度升高22℃。Au-Ag-Au nanorods经过磁性材料rGO-Fe3O4组装后,杀菌效果增强,30 μg mL-1的rGO-Fe3O4-Au-Ag-Au纳米复合材料在经磁分离后,近红外光照射20 min达到100%的杀菌效果。纳米复合材料rGO-Fe3O4-Au-Ag-Au能够有效的将磁分离与光热杀菌相结合,在提高杀菌效率、减少贵金属纳米棒用量的同时,还能将细菌从水体样本中分离去除。整个细菌捕获分离和灭活过程操作简单可行。
[Abstract]:Inorganic nanomaterials exhibit excellent physical and chemical properties different from conventional materials in the light, electricity, magnetism and sound, because of their gradual reduction in dimensions. The deep integration of inorganic nanomaterials with biological and chemical analysis technology has become a frontier and hot issue in the field of biomedical analysis and environmental analysis at home and abroad. However, the present large amount of research has been made in the field of biomedical analysis and environmental analysis. Some sensors based on nanomaterials are still in the laboratory research stage because of their high consumption, poor anti-interference ability and instability. Meanwhile, the controllable release of nanoparticles loaded with nanomaterials needs to be studied. Based on the above background, this paper aims to introduce nanomaterials into chemical / biological sensors. As well as drug loading and controlled drug release: (1) combining the excellent optical properties of inorganic nanomaterials, designing nano probes with purpose, building an analytical method with good anti-interference ability, good stability and high sensitivity, applied to the analysis of biological and environmental samples, and (2) design and synthesis of multi-functional nanocomposites. In order to realize the load and controllable release of drugs, improve the efficiency of bactericidal treatment, reduce or avoid the use of broad-spectrum antibiotics, the tissue damage caused by physical sterilization, or the loss of nutrients in the water. The first chapter of the paper briefly introduces the types, physical and chemical properties and applications of inorganic nanomaterials. The composition, classification, optical properties and applications in biological and environmental analysis of two inorganic nanomaterials of nanocrystalline and gold nanorods. The second chapter of the paper synthesizes the quantum dots (NAC-QDs) with N- acetyl -L- cysteine as the stabilizer and the Luo Danming 6G derivative (R6G-D) that have specific response to mercury, and the infrared spectra of them are carried out. Based on the fluorescence response of NAC-QDs and R6G-D to the fluorescence of mercury ions and the overlap of their absorption / emission spectra, a NAC-QDs/R6G-D fluorescence resonance energy transfer system was established to detect the ratio fluorescence of mercury and the quantitative detection of mercury ions in standard samples and environmental water samples. The linear range of the detection of mercury was 5-250. The precision is 3.2% (175 g L-1, n=11) and the detection limit is 0.75 mu g L-1 (3 Sigma /s, n=11). The FRET sensing system can change the chromatic aberration with the concentration of mercury ions in the concentration range of 0-250 mu g mL-1, and can be semi quantitative visual detection of mercury ions. The discriminability can reach the ratio of 50 mu g. The platform has high sensitivity, good selectivity, and the detection process is simple and the result is accurate. The method can realize the accurate quantitative detection and visual semi quantitative detection of the environment and biological samples in the environment and biological samples under the condition of reducing the cost. The third chapter of the paper synthesizes the hollow silica nanoscale (HSKAurod) loaded with gold nanorods coated with kanamycin. The nano composite HSKAurod nanomaterials also possess the physical and thermal germicidal properties of the gold nanorods and the chemical bactericidal properties of kanamycin, which can carry out the drug load and can be carried out at the same time as the photothermal treatment. Controlled release. The hollow silica nanoscale was used as a drug carrier to load the broad-spectrum antimicrobial kanamycin, and the gold nanorods were coated on the surface of nanoscale as near infrared photothermal response materials to avoid the leakage of kanamycin load. The bactericidal properties of HSKAurod nanocomposites were studied by selecting E. coli BL21 as the target strain. Energy and bactericidal mechanism.HSKAurod nanocapsts have high photothermal conversion efficiency. 10mgmL-1 materials can increase the environmental temperature from 21 C to 50.HSKAurod nanoscale rapidly in near infrared light (785 nm), and the bactericidal properties of the 10 mg mL-1 HSKAurod nanoscale can be killed by 20 min after near infrared light irradiation. The death rate almost reached 100%., which indicates that the combination of physical sterilization and chemical sterilization can produce synergistic effect of bactericidal, effectively reduce the dosage of antibacterials, shorten the time of photothermal treatment, reduce the biological tissue damage caused by physical sterilization and.HSKAurod nanoscale, which has the potential to replace the traditional germicide, which is applied to clinical sterilization. The fourth chapter of the thesis designs and synthesizes the bimetallic shell shell gold silver nanorod (Au-Ag-Au nanorods) as the photothermal bactericidal material of the near infrared light zone,.Au-Ag-Au nanorods outer layer gold shell which can be ablated in the near infrared light to expose the silver shell in the middle, so as to realize the controllable release of the antibacterial silver shell /Ag+. This combination of photothermal sterilization of the outer layer gold shell and the controllable release of /Ag+ in the intermediate silver shell gives the excellent bactericidal performance of Au-Ag-Au nanorods. The bactericidal and bactericidal mechanism of the bimetallic shell shell nanorods are studied by using the Escherichia coli E. coli 0157:H7 as a template, and the shell gold under low power near infrared laser irradiation. The pAu-Ag-Au nanorods (44 C) shows the photothermal conversion performance superior to the gold silver nanorods (Au-Ag nanorods, 39 C). At the same time, the nuclear shell shell structure of the bimetal nanorods has a better chemical stability than the Au-Ag nanorods, and the absorption spectrum of Au-Ag-Au nanorods is stable in 16 days, and the absorbance does not change the Au-Ag-Au Na of.10 micron gmL-1. Norods can make the temperature of the surrounding solution reach 44 centigrade by irradiation of 10 min near infrared light, and the killing rate of E. coli 0157:H7 of Escherichia coli to 100%. bimetallic core and shell Au-Ag-Au nanorods for sterilization at 20 min near infrared light, which successfully avoids the use of broad-spectrum antibiotics, reduces the amount of material and improves the sterilization efficiency. At the same time, the use of low power near infrared light reduces the damage to biological tissue. The above properties make the bimetallic shell shell Au-Ag-Au nanorods a new kind of nano photothermal conversion material for biomedical research in vivo. The fifth chapter of the thesis is first combined with the one step method to the magnetic rGO-Fe3O4-PEI nanometers with positive electricity. Materials, and a large number of bimetallic core shell shell Au-Ag-Au nanorods loaded on the reduced graphene oxide film layer, and the rGO-Fe3O4-Au-Ag-Au nanocomposites obtained are used as near infrared photothermal materials for the capture, separation and killing of bacteria. The nano composite material to the bacteria is studied with the Escherichia coli E. coli O157:H7 as the target strain. The capture ability, magnetic separation, photothermal conversion efficiency and the assembly of.RGO-Fe3O4 magnetic materials make rGO-Fe3O4-Au-Ag-Au nanocomposites able to identify and capture bacteria at the same time of near infrared photothermal sterilization. The rGO-Fe3O4-Au-Ag-Au of O30 mu gmL-1 from the water samples is 10 mmin under the action of the magnetic field. Within 100%, the E. coli O157:H7 with a concentration of 1 * 108 CFU mL-1 was identified and the magnetic separation from the solution was removed from the solution to remove the.RGO-Fe3O4 itself with a certain photothermal conversion performance, plus the excellent thermal conductivity of the graphene material, and the photothermal transfer efficiency of the nanocomposites was effectively improved after the assembly, and the rGO-Fe3O4-Au-Ag-Au of 25 mu g mL-1 was found. The near infrared radiation of 10 min can increase the temperature of the surrounding solution and increase the bactericidal effect after the.Au-Ag-Au nanorods of the magnetic material rGO-Fe3O4 is assembled by the magnetic material rGO-Fe3O4. The 30 mu g mL-1 nano composite material can achieve 100% bactericidal effect after the magnetic separation, and the near infrared light is irradiated by 20 min. The nano composite material rGO-Fe3O4-Au-Ag-Au can be obtained. It is effective to combine the magnetic separation with the photothermal sterilization to improve the bactericidal efficiency and reduce the amount of the noble metal nanorods, but can also separate the bacteria from the water samples. The operation of the whole bacteria capture separation and inactivation process is simple and feasible.
【学位授予单位】：东北大学
【学位级别】：博士
【学位授予年份】：2015
【分类号】：TB383.1

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