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研究生：	胡國威 Hu, Kuo-Wei
論文名稱：	發展金和釓為主之奈米粒子應用於生物醫學治療與診斷:熱治療，磁共振造影，非線性光學影像 Development of Gold- and Gadolinium-based Nanomaterials in Biological Diagnostic and Therapeutic Applications: Hyperthermia Therapy, MRI, Nonlinear Optics Microscopy
指導教授：	葉晨聖 Yeh, Chen-Sheng
學位類別：	博士 Doctor
系所名稱：	理學院 - 化學系 Department of Chemistry
論文出版年：	2010
畢業學年度：	98
語文別：	中文
論文頁數：	87
中文關鍵詞：	複合材料、顯影劑、光熱治療
外文關鍵詞：	hybrid material, contrast agent, photothermal therapy
相關次數：	點閱：74 下載：5
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由於奈米材料在縮小化後出現的嶄新特性以及尺度與生物分子相近，使得它們相當具有潛力成為新世代生醫領域應用的候選材料。近年來大量而廣泛的工作投入於材料的合成、控制組裝與修飾，無機奈米材料在生物醫學科技相關的應用在近年非常迅速的發展。許多生物科技應用的研究成果都證實了奈米材料的潛力，尤其在作為高敏感度以及具有選擇性的感應器或反應器時。高效能的奈米材料可有效降低對生物體注入試劑之劑量以及降低外來幅射輸出量的有效門檻並降低其可能帶來的副作用。此外，將具有不同性質且高效能之奈米粒子當作建構基材進一步人工或自行組裝為具有多重功能的單一材料也在近期逐漸被實現，這些成果大幅地展現奈米科技在生醫科技巨大的潛力。因此，具有高效能的奈米材料與多功能性的複合性材料不管在此刻還是將來都是相當具有研發潛力的一大方向。
本研究主要著重在發展具有治療與診斷效果之無機奈米材料並評估這些材料在實際應用的潛力。在本研究中我們合成了四種複合奈米材料，成分包含了金、銀、釓、矽等無機物，這些材料具有各自獨特的性質可提高顯影效果或是治療效果，分別為具有光熱治療效果，可作為光敏劑的金銀樹枝狀奈米結構；同時可作為光敏劑與可提升磁共振造影對比訊號的Gd2O(CO3)‧H2O/SiO2/金奈米殼層複合材料；同時可作為光敏劑與可提升非線性光學顯影對比的金奈米棒-金銀合金殼奈米結構；同時可提升磁共振造影對比訊號與可作為抗癌藥物傳遞載體的DOTA-釓錯合物接合之二氧化矽奈米管。除了分析與鑑定每個材料的成分與特性外，我們對所有的材料都進行了細胞毒性的測試以確定它們在生醫應用的適用性，然後從得到的結果評估材料的效能。結果顯示：金銀樹枝狀奈米結構具有光熱轉換的能力，且所需的輻射量比已被證實可作為光熱治療劑的金奈米棒還要低；Gd2O(CO3)‧H2O/SiO2/金奈米殼層複合材料不僅可提升磁共振造影信號，還可作為光熱治療劑，藉由精確地調控可以得到在兩者均有最佳表現的條件；金奈米棒-金銀合金殼奈米結構可同時作為光敏劑與提升非線性光學顯影對比，其效能比已被報導過的金奈米棒高；DOTA-釓錯合物接合之二氧化矽奈米管可提升水分子質子縱向弛緩速率，效能比DOTA-釓錯合物高數倍，此外，可同時作為藥物傳遞載體，將裝填的阿黴素導入癌細胞進行治療。

The properties arising from nanoscale effects and their comparable size to biomolecules have made nanomaterials act as potential candidates in next-generation biomedical applications. Progress in utilizing inorganic nanoparticles for biomedical applications has advanced rapidly due to the extensive amount of work done in the synthesis and modification of the materials. Nanomaterials have been proved promising in a wide range of biotechnological applications, especially as highly sensitive and selective sensors and reactors. Nanomaterials with high performance are feasible to significantly lower the dosage of external agents to bio-system and the threshold of radiation input, and then eliminate the possible side effect. Additionally, works related to the combination of different nanomaterials with a variety of properties and high performances have been reported in recent years, these results show that the nanomaterials exhibit great potential in biomedical applications. Therefore, developing nanomaterials with high efficacy and hybrid materials with multi-functionality is a highly promising subject regardless of now or future.
This study is aimed to the development of inorganic nanomaterials exhibiting the ability on diagnostics and therapeutics and the evaluation on their potential as applying to biotechnological field. We have fabricated four kinds of nano-scale hybrid materials containing inorganic species such as gold、silver、gadolinium or silicon. These materials include: AuxAg1-x dendrites, which converse the near-infrared radiation energy to heat effectively；Gd2O(CO3)‧H2O/SiO2/Au hybrid particles, which can be simultaneously served as magnetic resonance contrast agents and photothermal reactors；Au NR-in-shell nanostructures, which not only augment the nonlinear processes yield but also decrease the radiation threshold for the hyperthermia cancer therapy; DOTA-Gd complexes grafted silica nanotubes, which significantly enhance the relaxivity compared to DOTA-Gd complexes and can aslo be served as carriers for drug delivery to cancer cells. The results show several points as following : AuxAg1-x dendrites can effectively transfer the photon energy into heat, the required radiation for photothermal cancer therapy is significantly lower than that required for gold nanorods；Gd2O(CO3)‧H2O/SiO2/Au hybrid particles not only augment magnetic resonance signals but also can be served as photothermal therapeutic agents. By precise adjustment the optimal performance on magnetic resonance imaging and photothermal cancer therapy can be obtained；Au NR-in-shell nanostructures can simultaneously act as photothermal therapeutic agnets and nonlinear optic microscopy contrast agents, and they show higher efficacy than that of gold nanorods on both applications；DOTA-Gd complexes grafted silica nanotubes augment the longitudinal relaxation rate on protons of water molecules and exhibit higher efficiency than pure DOTA-Gd complexes for several times. On the other hand, they can also act as carriers for delivering doxorubicin molecules into HeLa cancer cells to induce therapeutic effect.

第一章	緒論                                        1
1奈米材料科技 (Nanomaterial Technology)...........................................................1
2貴重金屬奈米粒子的製備......................................................................................2
2.1 氧化還原反應..............................................................................................2
2.2 伽凡尼取代反應..........................................................................................3
3貴重金屬奈米粒子的特性......................................................................................4
3.1 體積效應......................................................................................................4
3.2 表面積效應..................................................................................................5
3.3 量子侷限效應..............................................................................................6
3.4 金屬奈米粒子獨特的光學特性..................................................................7
4 貴重金屬奈米粒子的穩定性.................................................................................8
5 奈米科技與生物醫學的結合.................................................................................9
6 金屬奈米粒子應用於癌細胞治療.......................................................................11
6.1 典型癌症治療方式....................................................................................11
6.2 光熱癌細胞治療法....................................................................................12
6.3 適用於光熱治療方法之金奈米材料........................................................13
6.3.1 金奈米粒子.....................................................................................13
6.3.2 金奈米棒(Gold Nanorod)...............................................................14
6.3.3 金核殼奈米結構(Gold Nanoshell).................................................14
6.3.4 籠狀金奈米結構(Gold Nanocage).................................................15
7 金屬奈米粒子應用於生醫診斷影像偵測技術...................................................16
7.1 比色法生物檢測........................................................................................16
7.2 非線性光學顯微診斷技術........................................................................17
7.2.1 非線性光學理論.............................................................................18
7.2.2 可提升非線性光學影像對比之材料.............................................19
7.3 磁共振造影技術(Magnetic Resonance Imaging)......................................20
7.3.1 磁共振造影原理.............................................................................20
7.3.2 顯影劑.............................................................................................21
7.3.3 新興奈米材料顯影劑.....................................................................22
8	奈米科技應用於藥物傳遞..................................................................................23
8.1 藥物傳遞系統............................................................................................23
8.2 藥物傳遞系統之載體類型........................................................................23
9 多功能複合型奈米材料.......................................................................................24

第二章	金銀樹枝狀奈米結構於光熱治療之應用         26
1 研究動機與目的...................................................................................................27
2 實驗部分...............................................................................................................28
2.1 藥品............................................................................................................28
2.2製備銀樹枝狀奈米結構.............................................................................28
2.3製備中空型金銀樹枝狀奈米結構.............................................................29
2.4中空型金銀樹枝狀奈米結構表面修飾抗體.............................................29
2.5 合成長寬比為4之金奈米棒.....................................................................29
2.6 光熱治療實驗............................................................................................29
2.7 儀器設備....................................................................................................30
3 結果與討論...........................................................................................................30
3.1 電子顯微鏡鑑定銀與中空型金銀樹枝狀奈米結構型態........................30
3.2 各種金銀成分比例之材料光學特性........................................................32
3.3 細胞毒性測試結果....................................................................................33
3.4 體外肺癌細胞A549光熱治療實驗結果..................................................33
4 結論.......................................................................................................................37
第三章	具MRI顯影劑與光熱轉換性質之雙功能Gd2O(CO3)‧H2O/SiO2/金奈米殼層複合材料    39
1 研究動機與目的...................................................................................................40
2 實驗部分...............................................................................................................41
2.1 藥品............................................................................................................41
2.2 Gd2O(CO3)‧H2O球型粒子之合成...........................................................42
2.3 在Gd2O(CO3)‧H2O球型粒子表面包覆帶胺基的二氧化矽層.............42
2.4在包覆二氧化矽層的Gd2O(CO3)‧H2O球型粒子表面形成金奈米殼層..........................................................................................................................43
2.5 Gd2O(CO3)‧H2O/silica/Au複合型粒子表面修飾抗體anti-HER2.........43
2.6 儀器設備....................................................................................................44
3 結果與討論...........................................................................................................44
3.1 Gd2O(CO3)‧H2O/silica/Au複合型粒子之型態與成分鑑定....................44
3.2 不同金殼層厚度複合型粒子之光學性質................................................46
3.3 不同金殼層厚度複合型粒子之T1加權影像與遲緩率...........................47
3.4 不同金殼層厚度複合型粒子之細胞毒性測試........................................48
3.5 不同金殼層厚度複合型粒子之癌細胞光熱治療測試............................48
4 結論.......................................................................................................................51
第四章	金奈米棒-金銀合金殼奈米結構之光熱治療及非線性光學顯影之應用                             52
1 研究動機與目的...................................................................................................53
2 實驗部分...............................................................................................................53
2.1 藥品............................................................................................................54
2.2合成金奈米棒-金銀合金殼奈米結構........................................................54
2.3金奈米棒-金銀合金殼奈米結構表面修飾抗體anti-EGFR......................54
2.4儀器設備.....................................................................................................55
3 結果與討論...........................................................................................................55
3.1 金奈米棒-金銀合金殼奈米結構型態、成分與光學性質鑑定.................55
3.2 光熱轉換實驗............................................................................................60
3.3 體外細胞毒性測試....................................................................................61
3.4 肺癌細胞A549光熱治療測試..................................................................62
3.5 金奈米棒-金銀合金殼奈米結構之非線性光學誘導放射光譜...............64
3.6 金奈米棒-金銀合金殼奈米結構之非線性光學細胞影像.......................65
4 結論.......................................................................................................................67
第五章	二氧化矽奈米管接合DOTA-釓錯合物應用於磁共振造影與藥物傳遞之研究                       69
1 研究動機與目的...................................................................................................70
2 實驗部分...............................................................................................................71
2.1 藥品............................................................................................................71
2.2 氫氧化釓奈米棒(Gd(OH)3 nanorods)之合成...........................................71
2.3 製備表面包覆二氧化矽層之Gd(OH)3奈米棒........................................71
2.4二氧化矽層表面修飾上胺基.....................................................................72
2.5 製備表面接有釓錯合物之二氧化矽奈米管(Silica Nanotubes-DOTA-Gd，SNDG)............................................................................72
2.6 儀器設備....................................................................................................72
3 結果與討論...........................................................................................................73
3.1二氧化矽奈米管的製備、修飾與鑑定.......................................................73
3.2 SNDG材料細胞毒性測試..........................................................................75
3.3 SNDG材料之MR顯影效果......................................................................76
3.4 SNDG作為載體應用於藥物傳遞..............................................................77
4 結論.......................................................................................................................78
第六章	 參考文獻                                  79
                                    

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