石墨烯包覆对红细胞结构功能影响

发布时间：2018-06-14 09:53

本文选题：氧化石墨烯 + 功能化修饰　；参考：《北京化工大学》2017年硕士论文

【摘要】：为了得到生物相容性较好的氧化石墨烯(Graphene Oxide,GO)以扩展其在生物医学领域的应用,采用氧化还原法(Hummers法)以氧化石墨为原料合成了氧化石墨烯,采用了能谱(XPS)、显微镜(AFM、TEM)等表征手段对合成的氧化石墨烯的结构进行了表征,表征结果显示通过氧化还原法成功合成了厚度约为1nm的单层氧化石墨烯。对实验合成的氧化石墨烯进行还原和分散分别得到RGO、BSA分散的GO、Tween 80分散的GO,并对GO、RGO、BSA分散的GO和Tween分散的GO进行了对于血细胞的毒性的研究。合成的氧化石墨烯经过Tween 80、BSA等的分散和修饰后,改变其原有的表面结构,再与从ICR小鼠经血提取得到的红细胞共同孵育不同时间后,测定红细胞溶血率的改变;将孵育后的细胞置于NaCl的梯度溶液中,检测红细胞达到50%溶血对应的NaCl浓度,用此数值反映经石墨烯包裹后细胞脆性的变化石墨烯再进入细胞后对细胞进行包覆,本实验研究了不同表面修饰和分散的石墨烯再包覆红细胞后对红细胞的溶血率和脆性方面的影响。研究的结果如下,氧化石墨烯及其相关衍生物有着较好的生物相容性,在2 mg/ mL下均呈现了较低的诱发溶血的性能。经石墨烯包裹后,氧化石墨烯与实验中通过经过进行还原和分散操作的氧化石墨烯相比,实验中所经过还原分散的氧化石墨烯血液相容性较好即血细胞毒性较小;而还原与分散的氧化石墨烯相比,经Tween 80分散的氧化石墨烯溶血率都较低,血液相容性较好即血细胞毒性较小。红细胞的脆性主要受分散剂的影响,pf127分散的氧化石墨烯表现出了一定的降低红细胞脆性的保护作用,BSA分散的氧化石墨烯对红细胞脆性几乎没有影响。石墨烯的包裹,减小红细胞的变形率,显示了一定保护红细胞的作用。通过对不同尺寸不同浓度的氧化石墨烯对小鼠脾细胞的毒性研究,进行CCK8实验,得出结论32目的氧化石墨烯对细胞的毒性比200目和1000目的氧化石墨烯更大。氧化石墨烯的浓度和尺寸对小鼠脾细胞的毒性有关。在一定范围内,氧化石墨烯尺寸越大,毒性越小;在氧化石墨烯在尺寸在200目时,氧化石墨烯对细胞有一定的保护作用,细胞活性达到125%,表现出较好的生物相容性。通过进行流式细胞实验,可以得出浓度为5 mg/mL氧化石墨烯尺寸在1000目的氧化石墨烯能进行简单的细胞分选。200目的氧化石墨烯对于细胞筛选作用很小。尺寸为1000目时,随着氧化石墨烯的浓度提高,CD19 PE+细胞群荧光强度也逐渐提升,由此判断氧化石墨烯有增强PE荧光强度的性质,CD3e FITC+而细胞群荧光强度逐渐下降,由此判断氧化氧化石墨烯猝灭了FITC的荧光,从而降低了 FITC的荧光强度。
[Abstract]:In order to obtain graphene oxide with good biocompatibility to expand its application in biomedical field, graphene oxide was synthesized from graphite oxide by redox method. The structure of graphene oxide was characterized by means of energy dispersive X-ray diffraction (XPS) and microscopical analysis. The results showed that graphene oxide with thickness of about 1nm was successfully synthesized by redox method. The RGOA BSA dispersed GOOTween80 was obtained by the reduction and dispersion of the synthesized graphene oxide. The cytotoxicity of GORGOBSA dispersed go and Tween-dispersed go to blood cells was studied. The synthesized graphene oxide was dispersed and modified by Tween80BSA, and the original surface structure was changed. After incubating with the red blood cells extracted from ICR mice for different time, the hemolysis rate of red blood cells was measured. The incubated cells were placed in the gradient solution of NaCl to detect the concentration of NaCl corresponding to 50% hemolysis of the red blood cells. The values reflected the changes of the fragility of the cells after the cells were encapsulated by graphene, and then the cells were coated with graphene after entering the cells. The effects of different surface modification and dispersion of graphene on the hemolysis rate and fragility of red blood cells were studied. The results are as follows: graphene oxide and its related derivatives have good biocompatibility and exhibit low induced hemolysis at 2 mg/ mL. After the graphene was encapsulated, graphene oxide had better blood compatibility than the graphene oxide, which was reduced and dispersed by reduction and dispersion, that is, the hemocytotoxicity was less. Compared with the dispersed graphene oxide, the hemolysis rate of the dispersed graphene oxide by Tween80 was lower, and the blood compatibility was better than that of the dispersed graphene oxide. The brittleness of red blood cells was mainly affected by dispersant. Pf127 dispersed graphene oxide showed a protective effect on erythrocyte brittleness. BSA dispersed graphene oxide had little effect on erythrocyte brittleness. The encapsulation of graphene reduces the deformability of red blood cells and shows the protective effect of red blood cells. The toxicity of graphene oxide with different sizes and concentrations to spleen cells of mice was studied and CCK8 experiment was carried out. It was concluded that the cytotoxicity of graphene oxide 32 was greater than that of 200 mesh and 1000 purpose graphene oxide. The concentration and size of graphene oxide are related to the toxicity of spleen cells in mice. In a certain range, the larger the size of graphene oxide is, the less toxic it is, and when the size of graphene oxide is 200 mesh, graphene oxide has a certain protective effect on cells, and the cell activity reaches 125%, showing good biocompatibility. By flow cytometry, it was found that the concentration of 5 mg / mL graphene oxide could be used for simple cell sorting at the size of 1000 脳 10 ~ (-1) 路min ~ (-1) 路L ~ (-1) 路min ~ (-1). When the size was 1000 mesh, the fluorescence intensity of CD19PE cells increased with the increase of graphene oxide concentration. It was concluded that graphene oxide had the property of enhancing the fluorescence intensity of PE and the fluorescence intensity of CD3e FITC decreased gradually. It is concluded that graphene oxide quenches the fluorescence of FITC and decreases the fluorescence intensity of FITC.
【学位授予单位】：北京化工大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R318;O613.71

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