碘氧铋纳米材料对人类皮肤角质细胞的毒性研究

发布时间：2018-02-04 05:31

本文关键词： BiOI纳米材料人类皮肤角质细胞细胞毒性理化性质　出处：《太原理工大学》2015年硕士论文　论文类型：学位论文

【摘要】：碘氧铋(BiOI)作为一种新型半导体纳米材料，因其独特的层状电子结构和出众的光催化性能，得到了极大关注。随着BiOI纳米材料研究和应用的日益广泛，其在环境中暴露的机会也越来越多，然而到目前为止，纳米BiOI的生物效应方面的研究还处于一片空白，因此开展此项研究尤为迫切。不同纳米材料的理化性质往往不同，如粒径、形貌、表面性质及团聚状态等，而这些不同通常会影响纳米材料与细胞间的相互作用，进而影响纳米材料的毒性。细胞毒性评价采用体外细胞培养法，其优点是快速有效，并且在揭示材料毒性的产生及其影响因素和毒性机理等方面具有明显的优势。因此，在细胞水平上研究BiOI的体外细胞毒性作用，并探讨毒性机制，建立可靠的毒性评价的方法，为BiOI的生物效应研究提供重要的参考依据，这将对BiOI日后的广泛应用有着深远的意义。基于以上考虑，本文以人类皮肤角质细胞(HaCaT)为模型，研究了三种不同的BiOI纳米材料的细胞毒性，主要分为以下三个方面： 1.首先，以无水硝酸铋、碘化钾为原料，分别以水、乙醇及乙二醇为溶剂，通过水/溶剂热法，在温度100°C，时间12h的条件下，合成BiOI纳米材料。结果表明：分别以水、乙醇及乙二醇为溶剂制备的BiOI纳米材料：BiOI(EG)、BiOI(ETH)和BiOI(H2O)，形貌分别为微球型、花球型以及片状结构，这三种经典结构为后续开展BiOI纳米材料对HaCaT细胞的毒性实验提供了原料。其次，本实验探索了混合溶剂对BiOI制备的影响，结果表明溶剂中有水加入时，BiOI粒径增大，微结构也发生变化，由微球向花球状转变；另外，通过添加表面活性剂PVP以及改变PVP量来控制BiOI形貌的生长。结果可知，无论以水、乙醇及乙二醇哪种为溶剂，添加PVP，均可以减小BiOI粒径，以EG为研究对象，增加PVP的加入量，可以进一步减小所得BiOI的粒径。 2.采用体外细胞培养模型HaCaT细胞株，首次比较研究了形貌分别为微球型、花球型以及片状结构的三种BiOI纳米材料暴露24小时后的细胞毒性。结果发现：三种BiOI纳米材料均产生了一定的细胞毒性和细胞膜损伤，且毒性与浓度正相关，BiOI(H2O)显示出比其他两种更强的细胞毒性，给药浓度为0.05~10μg/mL时呈现低毒性，最高浓度100μg/mL时，细胞存活率仅有50%左右，细胞形态也发生明显变化，细胞间隙增大，贴壁细胞明显减少，悬浮细胞增多，细胞变圆，甚至出现空泡现象。 3.考查了三种BiOI纳米材料的物理化学性质，并通过与细胞生化指标（如细胞活性、膜损伤、氧化应激、线粒体膜电位、细胞周期、细胞凋亡）和荧光显微镜、电子显微镜技术相结合，确定影响细胞毒性差异的因素，深入分析了三种BiOI纳米材料（片状、微球状及花球状）的细胞毒性机制。结果表明其可能的毒性机制是：BiOI与细胞作用，依靠细胞膜的流动性进入细胞，破坏细胞膜的完整性，，攻击细胞核及线粒体等细胞器，造成细胞凋亡和周期阻滞。这种毒理学差异主要与三种BiOI在细胞的内摄量和活性氧聚集有关，BiOI的形貌、粒径和表面性质对细胞毒性有着至关重要的作用，片状BiOI(H2O)的水合粒径较小，较易以直接的物理穿刺接触方式损伤细胞膜，进入细胞造成氧化损伤，球状BiOI(EG)具有丰富表面羟基且水合粒径较大，主要造成细胞氧化应激。对BiOI的细胞毒性研究及毒性机制的探讨，为确认BiOI纳米材料的生物安全性评价提供一定的实验依据。
[Abstract]:Iodine bismuth oxide (BiOI) as a new type of semiconductor nano materials because of its unique layered electronic structure and photocatalytic performance outstanding, has been of great concern. With the research and application of BiOI nano materials more widely, its exposure to the environment are more and more opportunities, but so far, the study on biological effect of nano the BiOI area is still in a blank, so the research is particularly urgent.
The physicochemical properties of different nano materials are often different, such as particle size, morphology, surface properties and agglomeration, and these differences usually affect the interaction between nano material and cells, thereby affecting the toxicity of nanomaterials. Cytotoxicity evaluation using in vitro cell culture method, the utility model has the advantages of fast and effective, and has obvious advantage in production and its affecting factors reveal material toxicity and toxic mechanism. Therefore, the in vitro cytotoxic effect of BiOI on the cell level, and to explore the mechanisms of toxicity, toxicity evaluation method to build reliable, provide an important reference for the study of biological effects of BiOI, which will be widely used on BiOI days after have a far-reaching significance.
Based on the above considerations, we studied the cytotoxicity of three different BiOI nanomaterials based on human skin keratinocytes (HaCaT), which are mainly divided into the following three aspects.
1. first, anhydrous bismuth nitrate, potassium iodide as raw materials respectively with water, ethanol and ethylene glycol as solvent by solvothermal synthesis, at a temperature of 100 DEG C, time 12h, synthesis of BiOI nano materials. The results show that the BiOI nano materials with water, ethanol and ethylene glycol as solvent. Preparation: BiOI (EG), BiOI (ETH) and BiOI (H2O), the morphology of microspheres respectively, and curd sheet structure, the three classical structure provides raw materials to carry out follow-up toxicity experiment of BiOI nanoparticles to HaCaT cells. Secondly, this study explored the effect of mixed solvent on the preparation of BiOI the results showed that the solvent in water is added, the BiOI particle size increases, the micro structure changes, changes from spheres to globular flower; in addition, by adding surfactants PVP and PVP change to control the amount and morphology of BiOI growth. The results, both in water, ethanol and ethylene glycol as the solvent which, Adding PVP, the particle size of BiOI can be reduced, and EG is used as the research object. The addition of PVP can further reduce the particle size of the obtained BiOI.
2. by in vitro cell culture model of HaCaT cells for the first time, the comparative study of the morphology of microspheres respectively, cytotoxicity after 24 hours of exposure and curd sheet structure of three BiOI nano materials. The results showed that: three kinds of BiOI nano materials have certain cytotoxicity and cell membrane damage, and toxicity and concentration are related BiOI (H2O) showed stronger cytotoxicity than the other two, drug concentration was 0.05~10 g/mL showed low toxicity, the highest concentration of 100 g/mL, the cell survival rate is only about 50%, the cell morphology was changed obviously, cell gap increases, the adherent cells were significantly reduced, suspended cells increased. The cells became round, and even the cavitation phenomenon.
3. to examine the physical and chemical properties of three kinds of BiOI nano materials, and with the cell and biochemical indexes (such as cell activity, membrane damage, oxidative stress, mitochondrial membrane potential, cell cycle, apoptosis) and fluorescence microscopy, electron microscopy combined to determine the influence factors of cell toxicity differences, in-depth analysis of the three BiOI nano materials (sheet, microsphere and globose) cytotoxic mechanism. The results showed that the toxic mechanism is: BiOI and cell function, depending on the fluidity of the cell membrane into the cell, undermine the integrity of cell membrane, nucleus and mitochondria attack, causing cell cycle arrest and apoptosis. The toxicology the main difference with the three kinds of BiOI in cells within the intake and active oxygen accumulation, BiOI morphology, particle size and surface properties play an important role on the cell toxicity, lamellar BiOI (H2O) hydrate particle size Small, easy to puncture direct contact physical damage to the cell membrane into the cell, oxidative damage, spherical BiOI (EG) has abundant surface hydroxyl groups and hydration of large particle size mainly caused by oxidative stress. To investigate the cytotoxicity and mechanism of toxicity of BiOI, and provide experimental basis for validation of biological safety the evaluation of BiOI nano materials.

【学位授予单位】：太原理工大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：TB383.1;TQ135.32

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