利用光催化誘發氧化還原反應來毒殺癌細胞的研究在過去已被廣泛的探討，特別是奈米化的觸媒物質經由紫外光催化後的反應被證實可有效地抑制許多癌細胞的生長，但是這樣的反應模式是否可應用在口腔癌細胞抑制尚未被探討。
  為了提高TiO2光催化活性且嘗試應用於毒殺口腔癌細胞，本研究以有機成分和金屬成分對TiO2進行改質，分別為葉酸(Folic acid)、Pt及Ag。因文獻證實大部分癌細胞的表面有葉酸的接收者(Folic Receptor)，本研究利用化學吸附方式，將葉酸與TiO2進行接枝，並利用FT-IR及SEM 進行物化分析；金屬改質部分，利用金屬本身可以捕捉TiO2 光生電子的能力，可延長電子電洞再結合的時間，使經過金屬改質光觸媒比TiO2 具有更佳的光催化效果，另外亦會使晶種趨於優勢晶種銳鈦礦(Anatase)，並於生物實驗中使用MTT Assay測試細胞存活率及CDD細胞死亡機制。
  改質後經物化分析，改質後TiO2粒徑大小皆為20 nm 左右，並無明顯改變，於FT-IR分析上，FA/TiO2特性官能基確實有削減及轉移的趨勢，代表改質有成功；XRD分析上，Pt/TiO2和Ag/TiO2分別有1%的 Pt 和 Ag沉積，且Ag/TiO2晶種於改質後趨於優勢晶種銳鈦礦(Anatase)。
  細胞實驗上，FA/TiO2對於毒殺口腔癌細胞、加速細胞胞飲能力皆無顯著效果；Pt/TiO2和Ag/TiO2毒殺口腔癌細胞的效果優於未改質TiO2，於劑量濃度200μg/ml，皆有最佳效果，細胞存活率分別為 54% 和 27%，比未改質TiO2好上30% 和 53%；死亡機制方面，未改質TiO2及Ag/TiO2於高劑量濃度，細胞趨於細胞凋亡(Apoptosis)，其餘條件處理下皆趨於細胞壞死(Necrosis)。

The photo-catalytic induced oxidation-reduction reaction by using nano TiO2 materials with applied UV have been shown to be effectively in inhibiting growth of many cancer cells.  Whether such reaction mode can be applied in oral cancer suppression has not yet been explored.
This study focus on the feasibility study of the photocatalytic modified TiO2 nanoparticles induce apoptosis of human oral squamous cancer cells. The TiO2 nanoparticles modified processes include using chemical adsorption to modify TiO2 particles with folic acid and using photo-catalytic synthesis with Pt and Ag.  The FT-IR、XRD、SEM and EDS tests were used to confirmed the success of the modified TiO2 nanoparticles.  An MTT test was used to compare the apoptosis of human oral squamous cancer cells under various conditions of the oxidation-reduction reaction induced by the modified TiO2 nanoparticles.  This study concludes that no significant effects of the folic acid modified TiO2 in increasing the ability to accelerate cell endocytosis.  On the other hand, the Pt/TiO2 and Ag/TiO2 modified TiO2 perform better than the folic acid modified TiO2 in accelerating cell endocytosis at dose 200μg/ml.  The survival rate of cells incubated with TiO2 modified with Pt and Ag are decreased to 54%(Pt/TiO2) and 27%(Ag/TiO2) which improve 30% and 53% when compared with the non-modified TiO2 respectively.  An apoptosis kit assay test confirm that both unmodified TiO2 and Ag modified TiO2 can induce apoptosis of the cancer cells, the other modified TiO2 usually result in necrosis of the cancer cells.

目錄  
圖目錄	IV
表目錄	VII
第一章   前言	1
1-1 研究緣起	1
1-2 研究目的	2
1-3 研究內容	3
第二章  文獻回顧	4
2-1 光催化反應原理	4
2-1-1 半導體特性	4
2-1-2  光催化機制	5
2-1-3二氧化鈦特性與應用	9
2-1-4 影響二氧化鈦光催化反應之因素	14
2-2 光觸媒材料改質	16
2-2-1以金屬成分改質	16
2-2-2以非金屬成分改質	25
2-3 界面活性劑	29
2-3-1界面活性劑種類	29
2-3-2界面活性劑之原理	30
2-4 口腔癌細胞之治療	31
2-4-1 口腔癌細胞之生成與特性	31
2-4-2 常見口腔癌細胞之治療方式	31
2-4-3 癌細胞死亡機制探討	33
2-4-4以光觸媒程序處理口腔癌細胞之優點與限制	34
2-5國內外光觸媒程序處理癌細胞相關研究	35
第三章 實驗方法與材料	37
3-1 實驗方法	37
3-1-1 實驗架構	37
3-1-2 TiO2改質實驗條件	38
3-2 實驗藥品	44
3-3 實驗儀器	45
3-4 生物實驗分析方法	46
3-4-1 DMEM 培養基配製	46
3-4-2 PBS配製	46
3-4-3 癌細胞培養	47
3-4-3 MTT Assay 細胞存活率分析方法	49
3-4-4 SRB Assay 細胞存活率分析方法	51
3-4-5 Cell Death Detection ELISA	51
3-5 化學實驗分析方法	52
3-5-1  FE- SEM、EDX、XRD、FT-IR 物化性質分析	52
第四章、結果與討論	55
4-1未改質二氧化鈦用量對口腔癌細胞致死實驗	55
4-1-1 MTT Assay	55
4-1-2 SRB Assay	57
4-2 改質二氧化鈦物化性質分析	60
4-2-1 FTIR 分析	60
4-2-2 SEM/EDS粒徑與含量分析	62
4-2-3 XRD 晶相分析	65
4-3 改質二氧化鈦對口腔癌細胞致死實驗	71
4-3-1 胞飲時間對毒殺效率之影響	71
4-3-2 胞飲程度對毒殺效率之影響	73
4-3-3 以金屬改質二氧化鈦毒殺癌細胞實驗	75
4-3-4 晶相變化對口腔癌細胞存活率的影響	81
4-4 分散劑對於溶解度與癌細胞之影響	82
4-4-1 Triton X-100	82
4-4-2 SDS	83
4-5 二氧化鈦顆粒對於癌細胞死亡機制之探討	84
4-5-1 Cell Death Detection Apoptosis	84
第五章、結論與建議	86
參考文獻	89
圖目錄
圖2-1 光催化機制圖	6
圖2-2 光觸媒圖解	7
圖2-3常見之半導體之能階分布圖	9
圖2-4 二氧化鈦結構圖	11
圖2-5 親水性實驗圖	13
圖2-6二氧化鈦用途	13
圖2-7 界面活性劑分子圖	29
圖2-8 界面活性劑的基本結構圖	30
圖 3-1 實驗架構	37
圖 3-2 以葉酸改質二氧化鈦製備流程圖	39 
圖3-3 以金屬改質二氧化鈦製備流程	41
圖3-4 細胞實驗流程圖	43
圖 3-5 細胞計數器圖	49
圖3-6 MTT結構圖	50
圖4-1 二氧化鈦濃度與Ca922 口腔癌細胞致死率關係圖(n=3)	56
圖4-2 Non-UV MTT 顏色變化圖	57
圖4-3 UV-60-min MTT顏色變化圖	57
圖4-4 二氧化鈦濃度與Ca922 口腔癌細胞致死率關係圖(n=3)	58
圖4-5 Non-UV SRB 顏色變化圖	59
圖4-6 UV-60 min SRB 顏色變化圖	59
圖4-7 葉酸結構圖	60
圖 4-8 FA/TiO2 FT-IR 分析圖	61
圖 4-9 FA/TiO2 FT-IR 分析圖	61
圖 4-10 FA/TiO2 FT-IR 分析圖	62
圖4-11未改質二氧化鈦 SEM圖	63
圖4-12以葉酸改質二氧化鈦 SEM圖	63
圖4-13以Pt改質二氧化鈦 SEM圖	64
圖4-14 以 Ag改質二氧化鈦SEM圖	64
圖4-15 未改質TiO2 XRD 圖	66
圖4-16 Pt/TiO2 (0.05%) XRD圖	67
圖4-17 Pt/TiO2 (0.2%) XRD 圖	68
圖4-18 Ag/TiO2 (0.05%) XRD 圖	69
圖4-19 Ag/TiO2 (0.2%) XRD 圖	70
圖4-20 胞飲 1 hr 細胞存活率圖(n=4)	71
圖4-21 胞飲 24 hr 細胞存活率圖(n=4)	72
圖4-22 胞飲 1、24 hr 細胞存活率圖(n=4)	73
圖4-23 胞飲程度對細胞存活率圖(n=4)	74
圖4-24 Pt/TiO2 (0.05%) 細胞存活率圖(n=4)	75
圖4-25 Pt/TiO2 (0.2%) 細胞存活率圖(n=4)	76
圖4-26 Ag/TiO2 (0.05%) 細胞存活率圖(n=4)	77
圖4-27 Ag/TiO2 (0.2%) 細胞存活率圖(n=4)	77
圖4-28 不同金屬摻雜比例對細胞存活率之影響(n=4)	78
圖4-29 不同金屬摻雜比例對細胞存活率之影響(n=4)	79
圖4-30 不同金屬摻雜對細胞存活率之影響(n=4)	79
圖4-31 不同金屬摻雜對細胞存活率之影響(n=4)	80
圖 4-32 晶相比例對口腔癌細胞存活率的影響	81
圖 4-33 晶相比例對口腔癌細胞存活率的影響	82
圖4-34 TX-100 對細胞存活率之影響	83
圖4-35 SDS 對細胞存活率之影響	83
圖4-36 口腔癌細胞死亡機制(n=2)	85

表目錄
表 2-1 二氧化鈦晶相物化性質	10
表2-2 金屬改質類型	17
表2-3以貴重金屬改質TiO2相關研究	18
表2-4以過渡金屬改質TiO2相關研究	21
表2-5以複合半導體改質TiO2相關研究	23
表2-6 以葉酸改質相關文獻	25
表2-7以非金屬改質TiO2相關研究	27
表2-8常見界面活性劑	30
表2-9 國內外相關文獻整理	35
表 3-1 不同二氧化鈦與葉酸比例之配比	38
表3-2 實驗條件	42
表3-3 實驗藥品	44
表3-4 實驗儀器	45
表3-5 PBS配方表	47
表4-1以Pt、Ag 改質二氧化鈦 EDS 分析表	65

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