臺灣博碩士論文加值系統

本研究中，我們開發出一個可以吸收近紅外光而驅動光動力療法之複合材料，而在材料的合成上採用layer-by-layer (LBL)的策略，將不同化合物修飾在奈米粒子。首先我們利用具有能量上轉換之摻雜鑭系元素奈米粒子(upconversion nanoparticles; UCNP)作為複合材料的主體，接著使用檸檬酸鈉進行Ligand Exchange，將UCNP表面上的油酸分子置換成檸檬酸根，藉此將UCNP修飾為水溶性並且表面帶負電。接著利用靜電吸附力將表面帶有64個NH2的第四代G4 PAMAM 樹枝狀高分子固定在UCNP表面上。帶有負電荷的三重態光敏感劑Ce6，再藉由靜電作用力吸附在奈米粒子的最外圍，而得到具備光動力療法的複合材料。相較於一般化學修飾法，透過LBL靜電吸附法便能夠快速而方便地將材料製備出來。接著使用980nm近紅外雷射光照射UCNP複合材料，當產生能量上轉換而釋放出三種不同波長的螢光-紫外光(365nm)、藍光(460nm)、紅光(660nm)。這三種光就會激發吸附在材料表面的Ce6，當能量轉移給周邊的三重態氧分子產生單重態活性氧分子，進而達到光動力療法的目的。在細胞實驗中，發現複合材料沒有毒性而且當複合材料吸附在低濃度Ce6(0.25μM)並藉由紅光驅動光動力療法，其細胞毒殺效果可達62.4%而單一藥物-Ce6僅12%，所以透過複合材料能夠有效增強光動力療法療效。此外，當利用近紅外光雷射驅動光動力療法其細胞毒殺效果將近70%。在未來希望能夠將此複合材料進行動物實驗來測試實際抑制癌細胞的效果。

In the study, we successfully synthesized a hybrid material that can carry functional molecules through a layer-by-layer (LBL) absorption strategy to perform near-infrared light (NIR)-promoted photodynamic therapy (PDT). First, the NIR-triggered PDT system was based on the lanthanide-doped energy upconversion nanoparticles (UCNP); then, the hydrophobic UCNPs were turned into the carboxylate-functionalized UCNP by exchanging the oleic acid with citrate ligands. We then loaded the G4 PAMAM dendrimers onto UCNP surface by simple electrostatic interactions between those NH2 groups of dendrimers and the COOH groups of citrate ligands. Afterward, a photosensitizer of Chlorin e6 (Ce6) bearing negatively charged COOH groups was allowed to absorb onto the dendrimer-coated UCNP by electrostatic binding. The LBL strategy provides a convenient route to prepare the stable nanocomplexes. The lanthanide-doped UCNP can emit upconverted UV, blue and red light under NIR-light excitation, and the visible light will excite the Ce6 on the UCNPs surface, followed by the energy transfer between the triplet Ce6 and surrounding oxygen to produce toxic singlet-oxygen (1O2), thus achieving the NIR-triggered PDT. In vitro experiments show that the NIR promoted-PDT material is nontoxic in dark but kill approximate 62% of MCF-7 cells under 660-nm (red) laser excitation at low Ce6 concentrations (0.25μM). In sharp contrast, the pristine Ce6 only kills approximately 12% of cells under the same red-light exposure. Most importantly, approximate 70% of cells is killed under the same concentration upon 980-nm (NIR) laser excitation, suggesting that the NIR-triggered is more effective than conventional red-light triggered PDT using the hybrid UCNPs as the photosensitizers. We anticipate the hybrid materials will be promising toward in vivo PDT for cancer treatment.

目錄
1-1 研究背景 1
1-2 研究動機及方法 3
第二章 文獻回顧 6
2-1 Upconversion Nanoparticles (簡稱UCNP)介紹 6
2-1-2上轉換機制 11
2-1-3水溶性UCNP之表面修飾方式 12
2-2 光動力療法(Photodynamic therapy簡稱PDT) 18
2-2-1光動力療法發展 18
2-2-2光動力療法機制 19
2-3樹枝狀高分子PAMAM dendrimer 21
2-3-1樹枝狀高分子合成 22
2-3-2樹枝狀高分子特性 23
2-4材料設計概念 24
第三章 實驗方法 27
3-1 實驗藥品 27
3-2 分析儀器 29
3-3 NaYF4:0.5%Tm/20%Yb (core) 奈米粒子之合成 31
3-4 NaYF4:Tm,Yb@ NaYF4 (core / shell)奈米粒子之合成 33
3-5 複合材料Ce6-G4-UCNP 製備 35
3-5-1水溶性COOH-UCNP製備 - Lemieux-von Rudloff 氧化法 35
3-5-2水溶性COOH-UCNP製備 –臭氧氧化法 36
3-5-3 水溶性 citrate-UCNP 製備 – Ligand Exchange 38
3-5-4 Layer by Layer – G4-Citrate-UCNP合成 39
3-5-5 Layer by Layer – Ce6-G4-Citrate-UCNP合成 40
3-6海螢螢光素分子單態氧偵測實驗 41
3-6-1單一藥物Ce6 -照射紫外光、藍光、紅光之單態氧偵測實驗 41
3-6-2單一藥物Ce6-增加溶劑含氧量並照光之單態氧偵測實驗 41
3-6-3單一藥物Ce6 -照射複合光源-白燈之單態氧偵測實驗 41
3-6-4複合材料照射紅光與近紅外光之單態氧偵測實驗 41
3-7 細胞光動力療法測試與細胞存活率測試 42
第四章 結果與討論 43
4-1 NaYF4:Tm,Yb@ NaYF4(core/shell)奈米粒子合成與鑑定 43
4-2複合材料Ce6-G4-UCNP合成與鑑定 58
4-2-1水溶性COOH-UCNP製備 –Lemieux-von Rudloff氧化法 58
4-2-2水溶性COOH-UCNP製備 - 臭氧氧化法 61
4-2-3 ligand exchange –citrate-UCNP合成與鑑定 65
4-2-4 Layer-by-Layer G4-citrate-UCNP 合成與鑑定 77
4-2-5 Layer-by-Layer Ce6-G4-Citrate-UCNP 合成與鑑定 87
4-3海螢螢光素分子單態氧偵測實驗 94
4-3-1單一藥物Ce6 -照射紫外光、藍光、紅光之單態氧偵測實驗 97
4-3-2單一藥物Ce6-增加溶劑含氧量並照光之單態氧偵測實驗 100
4-3-3單一藥物Ce6 -照射複合光源-白燈之單態氧偵測實驗 102
4-3-4單一藥物Ce6- 照射紅光與近紅外光之單態氧偵測實驗 104
4-3-5複合材料照射紅光與近紅外光之單態氧偵測實驗 106
4-4 細胞光動力療法測試 109
4-4-1 單一藥物Ce6之光動力療法測試 109
4-4-2 複合材料之光動力療法測試 112
第五章 結論 115
第六章 參考文獻 116
第七章 附錄 121

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