臺灣博碩士論文加值系統

青光眼目前為全球第二大的眼科疾病，其與高眼壓以及氧化壓力有關。青光眼治療方式十分繁瑣複雜且治療過程漫長，而在所有青光眼的治療方式當中，最常使用的治療方式是眼藥水，然而在眼藥水藥物遞送系統中有許多眼內屏障，而使眼藥水的生物利用度低導致治療效果並不顯著。鈰奈米粒子已廣泛被使用在生醫領域，因其有清除自由基的能力期許可降低青光眼造成氧化壓力。本論文用製備(1)具抗氧化能力且(2)中空之中空鈰奈米粒子(Hollow Cerium nanoparticles, hCe NPs)以提升藥物載量，並(3)將奈米粒子表面功能化接枝殼聚糖以打開細胞緊密連接增加生物利用率以及(4)將奈米粒子表面功能化有標靶功能的ZM241385分子。首先中空粒子合成是藉由模板法，以軟模板矽奈米粒子(Silica nanoparticles, Si NPs)吸附鈰奈米粒子(Cerium nanoparticles, CeNPs)後，並利用強鹼蝕刻以除去軟模板，而合成出中空鈰奈米粒子，再藉由表面修飾PEG以利後續功能化反應，目前已有成功利用化學反應接枝上殼聚糖(Chitosan, CS)以及ZM241385(ZM)。並利用材料分析，鑑定奈米粒子合成過程。在細胞實驗當中，可看出合成的奈米粒子在一定濃度下對細胞有良好生物相容性。在動物實驗治療青光眼的當中，最後開發的hCe-PEG-CS/ZM也有最大的治療效果。

Glaucoma is currently the second largest eye disease in the world and is associated with high intraocular pressure and oxidative stress. Glaucoma treatment is very cumbersome and complicated. The most commonly used treatment is eye drops. However, there are many intraocular barriers in eye , and the low utilization rate results in a non-significant treatment effect. Therefore, in this study, a glaucoma eyedrop with high bioavailability and versatility was developed. In this paper, the preparation (1) antioxidant capacity and (2) Hollow Cerium nanoparticles (hCe NPs) to increase the drug load, and (3) functionalized surface grafting of nanoparticles. The glycans are ZM241385 molecules that open the tight junctions of cells to increase bioavailability and (4) functionalize the surface of nanoparticles with targeted functions. First, the synthesis of hollow particles is carried out by template method, after adsorbing cerium nanoparticles (Cerium nanoparticles, CeNPs) with soft template nano particles (Si NPs), and etching with strong alkali to remove soft template. Hollow tantalum nanoparticles, and then surface modification PEG for subsequent functionalization, have been successfully grafted with chitosan (Chitosan, CS) and ZM241385 (ZM). And use material analysis to identify the synthesis process of nanoparticle. In the cell experiments, it can be seen that the synthesized nanoparticles have good biocompatibility to the cells at a certain concentration. Among the animal experiments for the treatment of glaucoma, the last developed hCe-PEG-CS/ZM also has the greatest therapeutic effect.

目錄
指導教授推薦書
口試委員會審定書
致謝........................................................................................................................................... iii
中文摘要................................................................................................................................... iv
Abstract ..................................................................................................................................... vi
目錄......................................................................................................................................... viii
圖目錄....................................................................................................................................... xi
表目錄..................................................................................................................................... xiii
第 1 章 諸論 .................................................................................................... - 1 -
1.1 研究動機 ......................................................................................................................... - 1 -
1.2研究目的 .......................................................................................................................... - 2 -
第 2 章 文獻回顧 ............................................................................................. - 4 -
2.1 眼睛介紹 ......................................................................................................................... - 4 -
2.2青光眼介紹 ...................................................................................................................... - 6 -
2.3 青光眼發病原因 ............................................................................................................. - 7 -
2.4 目前治療方式 ................................................................................................................. - 7 -
2.5 奈米粒子 ......................................................................................................................... - 8 -
2.6奈米鈰介紹 .................................................................................................................... - 10 -
2.7 多功能化材料 ............................................................................................................... - 12 -
2.8 材料功能化修飾 ........................................................................................................... - 13 -
2.9 新材料開發 ................................................................................................................... - 15 -
第 3 章 實驗設計與方法 ................................................................................. - 16 -
3.1 材料 ................................................................................................................................ - 16 -
3.2 實驗用設備 .................................................................................................................... - 18 -
3.3 實驗設計 ........................................................................................................................ - 19 -
3.3.1. 奈米矽球合成 ......................................................................................................... - 19 -
3.3.2 中空奈米鈰球合成 .................................................................................................. - 20 -
3.3.3 中空鈰球表面修飾 .................................................................................................. - 21 -
3.3.4 材料鑑定 .................................................................................................................. - 21 -
3.3.5 材料細胞相容性 ...................................................................................................... - 22 -
3.3.6 細胞ROS測定 ........................................................................................................ - 23 -
3.3.7 TEER&ZO-1 ............................................................................................................ - 24 -
3.3.8 體外載藥/釋放 .......................................................................................................... - 26 -
3.3.9 動物實驗 .................................................................................................................... - 27 -
3.4.1 統計分析 ................................................................................................................... - 27 -
第 4 章 結果與討論 ........................................................................................ - 28 -
4.1材料鑑定 ........................................................................................................................ - 28 -
4.1.1 UV ............................................................................................................................... - 28 -
4.1.2 TEM ............................................................................................................................ - 29 -
4.1.2.1 SiO2 .......................................................................................................................... - 29 -
4.1.2.2 SiO2@Ce .................................................................................................................. - 29 -
4.1.2.3 hCe ........................................................................................................................... - 31 -
4.1.3 Zeta&DLS ................................................................................................................ - 32 -
4.1.4 元素分析 .................................................................................................................. - 33 -
4.1.5 XRD ......................................................................................................................... - 35 -
4.1.6 FTIR ......................................................................................................................... - 36 -
4.1.7 功能化測定 ................................................................................................................ - 37 -
4.1.8體外載藥及釋放情況 ................................................................................................. - 39 -
4.2 細胞相容性 ................................................................................................................... - 40 -
4.2.1 體外相容性測試 ........................................................................................................ - 41 -
4.2.2 ROS測定 .................................................................................................................... - 45 -
x
4.2.3 ZO-1 ............................................................................................................................ - 47 -
4.2.4 TEER ........................................................................................................................... - 48 -
4.3.1 In vivo Study ............................................................................................................... - 49 -
第 5 章 結論 ................................................................................................... - 52 -
參考文獻 .................................................................................................................... - 54 -
圖目錄
圖2.1.1眼睛結構圖……………………………………………………………………… - 4 -
圖2.1.2房水流出通道…………………………………………………………………… - 5 -
圖2.1.3視神經細胞……………………………………………………………………… - 6 -
圖2.4.1藥物進入眼內障礙圖…………………………………………………………… - 8 -
圖2.6.1清除自由基循環圖……………………………………………………………… - 11 -
圖2.6.2奈米鈰應用領域………………………………………………………………… - 11 -
圖2.7.1眼內受體蛋白圖………………………………………………………………… - 13 -
圖2.7.2 ZM241385與A2A理論結合圖………………………………………………... - 13 -
圖2.8.1細胞緊密連接機制圖…………………………………………………………… - 15 -
圖3.3.1示意圖…………………………………………………………………………… - 19 -
圖3.3.2矽球合成反應式………………………………………………………………… - 20 -
圖3.3.3蝕刻形成中空鈰………………………………………………………………… - 21 -
圖 4.1.1 UV………………………………………………………………………………. - 28 -
圖4.1.2 SiNP……………………………………………………………………………... - 29 -
圖4.1.3吸附時間變化圖………………………………………………………………… - 30 -
圖4.1.4 中空鈰HR-TEM圖…………………………………………………………… - 31 -
圖4.1.5 SiNP EDS……………………………………………………………………….. - 33 -
圖4.1.6 hCe EDS………………………………………………………………………… - 34 -
圖4.1.7 hCe-PEG-CS/ZM EDS…………………………………………………………. - 34 -
圖4.1.8 材料XRD圖…………………………………………………………………… - 35 -
圖4.1.9 FT-IR……………………………………………………………………………. - 36 -
圖4.1.10 中空鈰濃度與DPPH相關性………………………………………………… - 38 -
圖4.1.11 各材料與DPPH相關性……………………………………………………… - 38 -
圖4.1.12 材料藥物封裝量………………………………………………………………..- 40 -
圖4.1.13 體外藥物釋放…………………………………………………………………. - 40 -
圖4.2.1 HLE的hCe 相容性…………………………………………………………….- 41 -
圖4.2.2 BCE的hCe 相容性……………………………………………………………. - 42 -
圖4.2.3 BCE的各組別材料顯微鏡圖…………………………………………………... - 43 -
圖4.2.4 BCE的各組別細胞MTS相容性測試………………………………………… - 43 -
圖4.2.5 BCE各組別材料相容性螢光圖………………………………………………... - 44 -
圖4.2.6 體外細胞ROS測試……………………………………………………………. - 46 -
圖4.2.7 ZO-1測試……………………………………………………………………….. - 47 -
圖4.2.8 細胞TEER(在第四個小時移除材料)………………………………………… - 48 -
圖4.3.1 兔子觀察………………………………………………………………………... - 49 -
圖4.3.2 兔子裂隙燈……………………………………………………………………... - 50 -
圖4.3.3 眼內壓分析……………………………………………………………………... - 51 -
表目錄
表4.1.1 Zeta&DLS ………………………………………………………………………- 32 -

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