利用氧化鋅奈米粒與抗癌藥物共沉積之氧化鋅紙進行癌症細胞毒性之研究

Cytotoxicity Test of Cancer Cells by Using ZnO Paper Prepared with Co-Deposition of ZnO Nanoparticles and Anti-Cancer Medicines

鄭瑋昌(Wei –Chang Zheng)

指導教授 ： 吳幼麟

國立暨南國際大學/科技學院/電機工程學系/碩士(2015年)

https://doi.org/10.6837/NCNU.2015.00286

摘要

本論文主要在探討以化學沉積法在紙張上透過共沉積的方式加入抗癌藥物與氧化鋅奈米粒(zinc oxide nanoparticles，ZnO NPs)相結合，製作成為含抗癌藥物之氧化鋅紙(zinc oxide paper，ZnO Paper)，並將其應用於癌症細胞之細胞活性檢測。我們探討不同沉積時間的共沉積抗癌藥物之氧化鋅紙之表面形貌及藥物變化，再將正常細胞與癌症細胞培養於氧化鋅紙張，透過MTT assay (3- (4,5-cimethylthiazol-2- yl)- 2,5-diphenyl tetrazolium bromide assay)分析以了解氧化鋅奈米粒是否有助於強化抗癌藥物對癌症細胞之毒性，並確認抗癌藥物是否可經由簡單之共沉積法加載於氧化鋅奈米粒上。
在分析上，我們先是以掃瞄式電子顯微鏡(scanning electron microscopy，SEM)觀察氧化鋅奈米顆粒於紙張上的沉積情形，並藉由傅立葉轉換紅外線光譜儀(Fourier Transform Infrared Spectrometer，FTIR)分析確認紙張上是否有抗癌藥物沉積。最後將細胞培養於氧化鋅紙張上，透過MTT assay分析並利用酵素結合免疫吸附法(enzyme-linked immunosobent assay，ELISA)檢測細胞活性。在本論文中，我們也利用氧化鋅紙擺放的位置，探討氧化鋅奈米粒直接與間接接觸對癌症細胞毒性的影響。
我們藉由SEM觀察到在纖維紙上沉積了結構清楚的ZnO奈米顆粒，再以FTIR分析可以看出紙張上的藥物會隨著時間變化其數量上是有變化的。我們以此證明化學沉積法可以成功將抗癌藥物成長於紙張上，並製出能強化癌症細胞毒性之共沉積抗癌藥物的氧化鋅紙。
我們將化學沉積法製作出來的共沉積抗癌藥物之氧化鋅紙，應用於癌症細胞之活性檢測。與未加抗癌藥物之氧化鋅紙相比較，我們發現共沉積抗癌藥物之氧檢化鋅紙對於癌症細胞之細胞毒性確有加強的功能，而與細胞做直接接觸會較間接接觸之細胞毒性稍強，但差異並不大。由我們的實驗結果可知，共沉積法可將抗癌藥物加載於氧化奈米粒上，而對於殺死癌症細胞有加成的作用，而此氧化鋅紙更可做為未來細胞毒性及新抗癌藥物開發之測試平台。

關鍵字

化學沉積法 ； 氧化鋅奈米顆粒 ； 氧化鋅紙 ； 共沉積 ； 細胞毒性 ； 細胞活性檢測

並列摘要

This paper forcuses on the chemical co-deposition of anti-cancer drugs and zinc oxide nanoparticles (ZnO NPs) on paper substrate to form the ZnO paper containing anti-cancer drug. The anti-cancer drug-containing ZnO paper was then used as a test plateform for cytotoxicity test for cancer cells. We investigated the changes of the surface morphology as well as the bondings of the ZnO NPs and anti-cancer drugs co-dpoisted paper substrate as a function of various co-deposition times. By using the MTT assay (3- (4,5-cimethylthiazol-2- yl)- 2,5-diphenyl tetrazolium bromide assay) analysis, we checked the cell viability of normal and cancer cells cultivated on the anti-cancer drug-containing ZnO paper. The purpose of this is to help us understand whether the ZnO NPs is useful for enhancing the cytotoxicity to cancer cells. In addition, we would like to confirm if the co-dposition can simply load the anti-cancer drugs on ZnO NPs.
In oder to confirm the anti-cancer drugs were properly loaded onto the ZnO NPs, the surface of the ZnO NPs and anti-cacer durgs co-deposited paper was observed by scanning electron microscope (SEM) and the bonding of which was detected by Fourier transform infrared spectroscopy (FTIR). Cell viability of cells cultivated on ZnO paper was analyzed by MTT assay in combining with enzyme-linked immunosorbent assay (Enzyme-linked immunosobent assay, ELISA). In this work, we also studied the cytotoxicity effect of direct contact and indirect contact of the ZnO NPs with the cells by placing the ZnO papers at different places in the culture plate.
ZnO NPs deposited on the fiber structure of the paper substrate can be observed clearly by SEM, and changes over time of the bonding of the anti-cncer drugs co-deposited with ZnO NPs can be discerned from the FTIR peaks. These results confirm that the anti-cancer drugs can be successfully loaded onto ZnO NPs through the proposed chemical co-deposition method.
We then applied the ZnO NPs and anti-cancer drugs co-deposited ZnO paper to the cytotoxicity test of cancer cells. By comparing with the ZnO paper without adding anti-cancer drugs, it is found that the anti-cancer drugs co-deposited ZnO paper does help in enhancing the cytotoxicity to the cancer cells, and the cytotoxicity effect of the ZnO paper contacted directly with the cancer cells is stronger than that contacted indirectly with the the cancer cells, though the difference is not obvious. placing the ZnO paper directly cotact with the cancer cells. In conclusion, our experimental results confirm that co-dposition of the anti-cancer drug and ZnO NPs can load the anti-cancer drug onto the ZnO NPs, and the resultant ZnO paper does enhance the apoptosis of cancer cells. The ZnO paper can be apparently used as a platform for cell cytotoxicity test and new anti-cancer drug development.

並列關鍵字

chemical reduction ； zinc oxide nanoparticle ； co-deposition ； zinc oxide paper ； cytotoxicity ； MTT assay test

參考文獻

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