近年來為了治療糖尿病，發展出最新技術為利用組織工程技術來重建人工胰臟組織。植入人體中的人工胰臟組織不但可以釋放足夠的胰島素，還能針對身體的需求適時反應並調控胰島素置放劑量，其工作效能可以達到接近原本胰臟的工作能力，本研究是將細胞被包圍在一個直徑約750μm的水膠球中，因為缺乏血管運輸養分與氧氣，細胞的代謝必須完全靠擴散作用完成，因此如何調控水膠球的大小，並且了解氧氣與代謝物質在膠球內之分佈狀況，對於是否能長期維持健康的細胞生長成為點當中要的關鍵因素。
將市面上普遍可購得到表面修飾羧基的聚苯乙烯奈米微球(Polystyrene Nanobeads)經過一系列的製程處理後將可得到奈米氧氣感測器(Nano Oxygen Particles, NOPs)，而本研究所採用氧氣感測螢光化學式為C72H48Cl2N6Ru，又稱做Ru(dpp)3Cl2，讓高度生物相容性的Pluronic F-127當作支架，將對氧氣有感測效果的Ru(dpp)3Cl2包附F-127的疏水中心，讓親水端和膠球上羧基酯化反應後鍵結，再將NOPs和細胞放在水膠的球狀結構內並使其正常工作，藉由觀察NOPs的螢光亮度可以偵測環境上的溶氧量，而氧氣環境對動物細胞的生理機制尤其重要，若能得知環境氧氣訊息將可以做進一步的應用，例如：將水膠球狀結構植入人體的皮下，胰島細胞將釋放出胰島素並滲出水膠球外，讓糖尿病患血液中血糖含量恢復正常，且NOPs為點狀奈米感測器，有著訊噪比高的優點，是一個具有潛力的研究。

In order to treat diabetes, the development of the latest technology is that scientists use tissue engineering techniques to rebuild artificial pancreas tissue in recent years. The implanted artificial pancreas tissue not only release enough insulin, but also timely response to the needs of the body and regulate the insulin dose. The efficiency can reach close to the original working capacity of the pancreas. We culture the cells inside a hydrogel sphere which the diameter is about 750μm. Metabolism of cells must be fully completed by diffusion because of lack of blood vessels which transport nutrients and oxygen. Therefore, controlling hydrogel spheres size and understanding the oxygen distribution within hydrogel spheres is an important key point for cell healthy growth.
In this research, we use nanopolystyrene beads to obtain nanoxygen particles (NOPs) after a series of processes. And then, we let NOPs and normal islet cells work together within a spherical hydrogel structure. We can detect dissolved oxygen of environments by observing fluorescence intensity of NOPs. Oxygen of environments is particularly important for the physiological mechanism of animal cells. If we get the information, it will be able to make further applications. For example : We could let hydrogel spheres implant subcutaneously. Islet cells release insulin which diffuses to out of spheres, so diabetics blood sugar levels back to normal. NOPs are a point-like nanosensors and have high signal to noise ratio. It’s a promising research.

總目錄
摘要 I
總目錄 III
圖目錄 V
表目錄 VIII
第一章 緒論 1
1.1 研究背景 1
1.2 研究動機 2
1.3 研究目的 4
第二章 文獻回顧 6
2.1 偵測多重性質之奈米感測器 6
2.2 氧氣螢光分子的應用 8
2.2.1 球形結構 8
2.2.2 細胞生理環境 13
2.3 水膠的應用 16
2.3.1 管狀水膠 16
2.3.2 球狀水膠 20
第三章 實驗設計、原理與步驟 27
3.1 實驗設計 27
3.2 實驗原理 28
3.3 實驗步驟 29
3.3.1 NOPs製作 29
3.3.2 無菌式電噴灑水膠球 31
3.4 實驗材料 32
3.4.1 聚苯乙烯(Polystyrene)[63] 32
3.4.2 螢光分子 33
3.4.3 水膠 36
3.5 實驗儀器 40
第四章 實驗結果與討論 44
4.1 利用FTIR判定酯基存在 44
4.1.1 控制變因：溫度 44
4.1.2 控制變因：加熱攪拌時間 46
4.2 NOPs在空氣中螢光校正 48
4.3 NOPs在不同氣體中之螢光回復性 50
4.4 NOPs在去離子水中螢光校正 52
4.4.1 imageJ螢光亮度分析 52
4.5 NOPs在水膠球中螢光校正 55
4.6 NOPs和Hela在水膠球中培養 58
4.7 NOPs在A549細胞內培養 66
第五章 結論 68
5.1 實驗設計 68
5.2 FTIR判定酯基是否存在 68
5.3 NOPs在空氣中螢光校正 68
5.4 NOPs在不同氣體中之螢光回復性 68
5.5 NOPs在去離子水中螢光校正 69
5.6 NOPs在水膠球中螢光校正 69
5.7 NOPs和Hela在水膠球中培養 69
5.8 NOPs在A549細胞內培養 70
第六章 未來工作 71
參考文獻 72

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