本文關(guān)鍵詞： BiOI納米材料 人類皮膚角質(zhì)細(xì)胞 細(xì)胞毒性 理化性質(zhì)　出處：《太原理工大學(xué)》2015年碩士論文　論文類型：學(xué)位論文

【摘要】：碘氧鉍(BiOI)作為一種新型半導(dǎo)體納米材料，因其獨(dú)特的層狀電子結(jié)構(gòu)和出眾的光催化性能，得到了極大關(guān)注。隨著BiOI納米材料研究和應(yīng)用的日益廣泛，其在環(huán)境中暴露的機(jī)會也越來越多，然而到目前為止，納米BiOI的生物效應(yīng)方面的研究還處于一片空白，因此開展此項研究尤為迫切。 不同納米材料的理化性質(zhì)往往不同，如粒徑、形貌、表面性質(zhì)及團(tuán)聚狀態(tài)等，而這些不同通常會影響納米材料與細(xì)胞間的相互作用，進(jìn)而影響納米材料的毒性。細(xì)胞毒性評價采用體外細(xì)胞培養(yǎng)法，其優(yōu)點(diǎn)是快速有效，并且在揭示材料毒性的產(chǎn)生及其影響因素和毒性機(jī)理等方面具有明顯的優(yōu)勢。因此，在細(xì)胞水平上研究BiOI的體外細(xì)胞毒性作用，并探討毒性機(jī)制，建立可靠的毒性評價的方法，為BiOI的生物效應(yīng)研究提供重要的參考依據(jù)，這將對BiOI日后的廣泛應(yīng)用有著深遠(yuǎn)的意義。 基于以上考慮，本文以人類皮膚角質(zhì)細(xì)胞(HaCaT)為模型，研究了三種不同的BiOI納米材料的細(xì)胞毒性，主要分為以下三個方面： 1.首先，以無水硝酸鉍、碘化鉀為原料，分別以水、乙醇及乙二醇為溶劑，通過水/溶劑熱法，在溫度100°C，時間12h的條件下，合成BiOI納米材料。結(jié)果表明：分別以水、乙醇及乙二醇為溶劑制備的BiOI納米材料：BiOI(EG)、BiOI(ETH)和BiOI(H2O)，形貌分別為微球型、花球型以及片狀結(jié)構(gòu)，這三種經(jīng)典結(jié)構(gòu)為后續(xù)開展BiOI納米材料對HaCaT細(xì)胞的毒性實(shí)驗(yàn)提供了原料。其次，本實(shí)驗(yàn)探索了混合溶劑對BiOI制備的影響，結(jié)果表明溶劑中有水加入時，BiOI粒徑增大，微結(jié)構(gòu)也發(fā)生變化，由微球向花球狀轉(zhuǎn)變；另外，通過添加表面活性劑PVP以及改變PVP量來控制BiOI形貌的生長。結(jié)果可知，無論以水、乙醇及乙二醇哪種為溶劑，添加PVP，均可以減小BiOI粒徑，以EG為研究對象，增加PVP的加入量，可以進(jìn)一步減小所得BiOI的粒徑。 2.采用體外細(xì)胞培養(yǎng)模型HaCaT細(xì)胞株，首次比較研究了形貌分別為微球型、花球型以及片狀結(jié)構(gòu)的三種BiOI納米材料暴露24小時后的細(xì)胞毒性。結(jié)果發(fā)現(xiàn)：三種BiOI納米材料均產(chǎn)生了一定的細(xì)胞毒性和細(xì)胞膜損傷，且毒性與濃度正相關(guān)，BiOI(H2O)顯示出比其他兩種更強(qiáng)的細(xì)胞毒性，給藥濃度為0.05~10μg/mL時呈現(xiàn)低毒性，最高濃度100μg/mL時，細(xì)胞存活率僅有50%左右，細(xì)胞形態(tài)也發(fā)生明顯變化，細(xì)胞間隙增大，貼壁細(xì)胞明顯減少，懸浮細(xì)胞增多，細(xì)胞變圓，甚至出現(xiàn)空泡現(xiàn)象。 3.考查了三種BiOI納米材料的物理化學(xué)性質(zhì)，并通過與細(xì)胞生化指標(biāo)（如細(xì)胞活性、膜損傷、氧化應(yīng)激、線粒體膜電位、細(xì)胞周期、細(xì)胞凋亡）和熒光顯微鏡、電子顯微鏡技術(shù)相結(jié)合，確定影響細(xì)胞毒性差異的因素，深入分析了三種BiOI納米材料（片狀、微球狀及花球狀）的細(xì)胞毒性機(jī)制。結(jié)果表明其可能的毒性機(jī)制是：BiOI與細(xì)胞作用，依靠細(xì)胞膜的流動性進(jìn)入細(xì)胞，破壞細(xì)胞膜的完整性，，攻擊細(xì)胞核及線粒體等細(xì)胞器，造成細(xì)胞凋亡和周期阻滯。這種毒理學(xué)差異主要與三種BiOI在細(xì)胞的內(nèi)攝量和活性氧聚集有關(guān)，BiOI的形貌、粒徑和表面性質(zhì)對細(xì)胞毒性有著至關(guān)重要的作用，片狀BiOI(H2O)的水合粒徑較小，較易以直接的物理穿刺接觸方式損傷細(xì)胞膜，進(jìn)入細(xì)胞造成氧化損傷，球狀BiOI(EG)具有豐富表面羥基且水合粒徑較大，主要造成細(xì)胞氧化應(yīng)激。對BiOI的細(xì)胞毒性研究及毒性機(jī)制的探討，為確認(rèn)BiOI納米材料的生物安全性評價提供一定的實(shí)驗(yàn)依據(jù)。
[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.

【學(xué)位授予單位】：太原理工大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2015
【分類號】：TB383.1;TQ135.32

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