二氧化鈦及氧化鋁奈米粒細胞毒性之研究

The Study of Cytotoxicity of TiO2 and Al2O3 Nanoparticles

張哲銘(Che-Ming Chang)

指導教授 ： 吳幼麟

共同指導教授 ： 林錦正(Chin-Cheng Lin)

國立暨南國際大學/科技學院/電機工程學系/碩士(2014年)

https://doi.org/10.6837/NCNU.2014.00315

摘要

偵測細胞與基板間的交互作用被視為檢測細胞特性的一種方法。在本論文中，我們試著利用原子力顯微鏡來觀察奈米粒子(Nanoparticles，NPs)對正常細胞與癌症細胞之細胞毒性的影響。在塗有3 - 胺丙基三乙氧基矽烷(3-aminopropyltriethoxysilane，γ-APTES)修飾的表面先做細胞的培養，再以原子力顯微鏡量測細胞移除後所留下印痕形貌印痕，並探討在添加不同參數的NPs作用下不同細胞與基板間交互作用的關係。

伴隨奈米技術的逐年進步，具有獨特的化學物理特性- 如極小的尺寸、表面反應、電荷特性、形狀和與介質的交互作用之奈米粒子被製造出來，並被廣泛的使用。同時，奈米粒子的某些新特性可能會導致不良的生物影響，增加奈米粒子的製造和使用容易造成人類環境暴露於奈米粒子，因此，奈米粒子的安全性與其對於細胞的毒性研究是極為重要的。

在本論文中，我們分別使用人胚肺成纤维细胞正常細胞WI-38、人類肺腺癌上皮細胞A549、人類肺癌細胞株H1299在不同奈米粒子Al2O3、TiO2與不同奈米粒子濃度(0μg/L、100 μg/L、500 μg/L、1000 μg/L)作用下，以及使用抗癌藥物Staurosproine、Iressa環境下來探討細胞與NPs在基板上留下的印痕關係。所有測試的細胞在不同條件下於γ-APTES表面培養24小時後予以移除，再利用原子力顯微鏡觀察細胞移除表面後的形貌，藉由此形貌來分析奈米粒子對細胞毒性之影響，同時也使用細胞存活率檢測(3-(4,5)-dimethylthiahiazo (-z-y1)-3,5-di- phenytetrazoliumromide，MTT)來印證細胞存活率與細胞印痕形貌印痕之間關係是否符合NPs對於細胞的毒害。

關鍵字

氧化鈦 ； 氧化鋁 ； 人類肺腺癌細胞 ； 人肺細胞 ； 原子力顯微鏡

並列摘要

Detection of cell-substrate interaction is one of the methods for understanding cells properties. In this thesis, we tried observe the effect of nanoparticles on the cytotoxicity of normal and cancer cells by measuring the post-cell-removal surface morphology profiles with atomic force microscopy (AFM). Both the normal and cancer cells were cultivated on 3-aminopropyltriethoxysilane(γ-APTES) surface with and without adding different amount of nanoparticles (Al2O3 and TiO2) into the culture mediums, then the difference of the post-cell-removal surface morphology profiles were measured and compared by AFM.

Due to the rapid progress in nanotechnology, many nanoparticles with unique chemical properties such as tiny size, surface reactivity, charge, shape, and media interactions, have been fabricated and widely used. However, NPs could lead to adverse biological effects with potential cytotoxicity. Increasing usage and manufacturing of NPs increases exposure probability of humen to NPs. Therefore, it is extremely important to realize the safety and toxicity of NPs.

In this thesis, human embryonic lung fibroblast cell WI-38, human lung adenocarcinoma epithelial cells A549, human lung cancer cell lines H1299, were investigated under the influences of NPs Al2O3 and TiO2 with different concentrations (0μg/L、100 μg/L、500 μg/L、1000 μg/L). Their effect on the normal and cancer cells when subjected to anti-cancer medicines Staurosporine and Iressa was also investigated. Living normal and cancer cells were cultivated on the γ-APTES modified SiO2 surface in culture mediums under different conditions, and the post-cell-removal surface morphology profiles were obtained after the cells were cultivated for 24 hours. The changes of the post-cell-removal surface morphology profiles were analyzed before and after adding the stimulus. In order to confirm the post-cell-removal surface morphology profile results, the conventional MTT tests were also conducted for the cytotoxicity of NPs.

並列關鍵字

Titanium oxide ； Aluminium oxide ； A549 ； H1299 ； WI38 ； Atomic Force microscopy

參考文獻

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