臺灣博碩士論文加值系統

本研究開發之上轉換奈米粒子可透過反應時間有效控制其尺寸，透過980 nm波段之近紅外光照射後可造成其吸收多個光子並發出綠色可見光，此一特性得以解決光動力治療時因可見光穿透力不足造成無法進行非侵入式治療之問題，因上轉換奈米粒子之激發光源具有足夠穿透深度和粒子本身低自體螢光等優點，因此相當適合應用於光動力治療領域。過去研究多將光動力治療應用於癌症，光動力治療是利用光敏劑受到可見光激發後，因為能量的轉移使細胞周圍的氧分子接收到轉移的能量而形成活性氧物種，活性氧物種便會對細胞造成傷害，最後導致光敏劑周遭之癌細胞凋亡。而本研究則結合上轉換奈米粒子與光動力治療應用於類澱粉蛋白之降解，是由於活性氧物種的產生可以使類澱粉蛋白產生氧化的效果，最後造成鍵結的斷裂，更突破了以往光動力學以及上轉換奈米粒子之應用。除此之外，本研究所開發之上轉換奈米粒子在經過矽殼以及玫瑰紅鈉鹽的表面修飾後進行分析，透過一步又一步的驗證更證實了本研究所合成之上轉換奈米粒子成功進行上述所提及之表面修飾且不影響本身之特性，不論是在晶相、螢光性能亦或是生產活性氧物種的觀察中皆具有良好的表現。不僅如此，於本研究的最後成功藉由石英晶體微天秤證實了上轉換奈米粒子確實可以有效切割類澱粉蛋白之序列，且在結果中可明顯觀察到序列是否斷裂之差異。

The upconversion nanoparticles developed by this research can effectively control its size through the reaction time. After being irradiated with near-infrared light in the 980 nm, it can cause it to absorb multiple photons and emit green visible light. The problem of non-invasive treatment cannot be performed due to insufficient visible light penetration, but the excitation light source of the upconversion nanoparticles has the advantages of sufficient penetration depth and low self-fluorescence of the particles, so it is very suitable for application in the field of photodynamic therapy. In the past, most researches applied photodynamic therapy to cancer. Photodynamic therapy uses a photosensitizer to be excited by visible light. Because of the energy transfer, the oxygen molecules around the cell receive the transferred energy to form reactive oxygen species, so the cells were damaged and eventually lead to apoptosis of the cancer cells around the photosensitizer. In this study, the combination of upconversion nanoparticles and photodynamic therapy was applied to the degradation of amyloid-beta, because the generation of reactive oxygen species can cause the oxidation effect of amyloid-beta, and finally cause the breakage of the peptide, which is more breakthrough than photodynamic therapy application before. In addition, the upconversion nanoparticles developed by us were analyzed after surface modification of the silicon shell and Rose Bengal, and the success of the synthesis of upconversion nanoparticles in this research was confirmed by step-by-step verification. The surface modification mentioned above does not affect its own characteristics, whether it is in the crystalline phase, the emitting performance, or the production of reactive oxygen species, all have good performance. Not only that, but our success at the end of this study also confirmed that the upconversion nanoparticles can effectively cleave the amyloid-beta sequence analyzed by quartz crystal micro-scale, and the difference in whether the sequence is broken can be clearly observed in the results.

目錄
摘 要 i
Abstract ii
致謝 iv
目錄 v
圖目錄 viii
表目錄 x
第1章 緒論 1
1.1 研究背景 1
1.2 研究動機及目的 3
第2章 文獻回顧 5
2.1 阿茲海默症 (Alzheimer’s Disease) 5
2.2 類澱粉前驅蛋白 (Amyloid precursor protein；APP) 6
2.3 β類澱粉蛋白 (Beta-amyloid；Aβ) 8
2.4 Tau蛋白 (Tau protein) 9
2.5 奈米材料原理及應用之介紹 10
2.6 近紅外光譜 (Near Infrared Spectrum, NIRS) 於生醫領域的重要性 11
2.7 上轉換奈米材料簡介 12
2.8 光子上轉換機制 13
2.9 鑭系元素電子軌域躍遷之介紹 15
2.10 上轉換奈米材料常見製備方法 16
2.11 材料表面修飾方法之比較 17
2.12 上轉換奈米材料之應用 20
第3章 材料與方法 23
3.1 實驗材料 23
3.2 實驗儀器 28
3.3 實驗架構 30
3.4 實驗方法及步驟 31
3.4.1 熱共沉澱法製備 NaYF4:Yb/Er 31
3.4.2 對 NaYF4:Yb/Er 進行矽殼之表面修飾 32
3.4.3 NaYF4:Yb/Er@SiO2 表面之胺基修飾 33
3.4.4 利用 Rose Bengal 進行表面修飾 33
3.4.5 同時進行 Rose Bengal 及 Amyloid beta 對材料的表面修飾 34
3.4.6 SEM試片製備 34
3.4.7 TEM試片製備 35
3.4.8 DCFH-DA測試 35
3.4.9 QCM測試 35
3.4.10 生物相容性測試 35
第4章 結果與討論 36
4.1 掃描式電子顯微鏡對 NaYF4:Yb/Er 表面形貌之觀察 37
4.2 穿透式顯微鏡對 NaYF4:Yb/Er@SiO2 之形貌觀察及比較 40
4.3 NaYF4:Yb/Er 與 NaYF4:Yb/Er@SiO2 之晶相分析鑑定 44
4.4 NaYF4:Yb/Er@SiO2 胺基含量之比較 46
4.5 上轉換奈米粒子發光性能之研究 47
4.6 上轉換奈米粒子之吸收光譜 50
4.7 DCFH-DA測試 51
4.8 Rose Bengal 功能之驗證 52
4.9 生物相容性測試 54
第5章 結論 56
參考文獻 58
圖目錄
圖 2 1. 類澱粉蛋白假說之致病機轉 [11] 6
圖 2 2. 類澱粉前驅蛋白形成Aβ之機制 7
圖 2 3. Aβ1-40 與 Aβ1-42 序列 8
圖 2 4. Tau protein 與過度磷酸化 Tau protein 之示意圖 9
圖 2 5. 近紅外光之吸收光譜圖 11
圖 2 6. 上轉換奈米粒子發光示意圖 12
圖 2 7. 光子上轉換之機制。(a) ESA;(b) ETU;(c) PA 14
圖 2 8. Yb3+與Er3+離子能階轉換過程 16
圖 2 9. 奈米材料常見表面修飾方法 19
圖 2 10. 活性氧物種。(a) 1O2;(b) H2O2;(c) OH· 21
圖 2 11. 上轉換奈米粒子激發 RB 示意圖 22
圖 3 1. 實驗架構圖 30
圖 3 2. 上轉換奈米粒子合成步驟 31
圖 3 3. 矽殼修飾示意圖 32
圖 3 4. 胺基修飾示意圖 33
圖 3 5. 光敏劑修飾示意圖 33
圖 3 6. 光敏劑及 Amyloid-beta 修飾示意圖 34
圖 4 1. NaYF4:Yb/Er 以不同時間合成之 SEM圖。 38
圖 4 2. NaYF4:Yb/Er 以不同時間合成之粒徑分布圖 39
圖 4 3. NaYF4:Yb/Er 元素分析之結果。 40
圖 4 4. NaYF4:Yb/Er@SiO2 與 NaYF4:Yb/Er之 TEM 比較圖 42
圖 4 5. NaYF4:Yb/Er@SiO2 之 TEM Mapping 44
圖 4 6. (a)NaYF4:Yb/Er 與 (b)NaYF4:Yb/Er@SiO2 之 XRD 比較圖 (c) JCPDS CARD:28-1192 45
圖 4 7. APTES 濃度對於胺基含量之比較 46
圖 4 8. UCNP 與 SiO2 之光譜比較圖 48
圖 4 9.帶有光敏劑之粒子與 SiO2 之光譜比較圖 49
圖 4 10. 修飾前後上轉換奈米粒子之吸收光譜 50
圖 4 11. DCFH-DA 氧化示意圖 51
圖 4 12. ROS 生成檢測 52
圖 4 13. QCM分析之步驟 53
圖 4 14. Rose Bengal 對 Aβ 造成氧化之驗證 54
圖 4 15.細胞存活率測試 55
表目錄
表4 1. 本實驗合成之上轉換奈米材料參數。 36

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