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研究生： 吳俊廷
Wu, Chun-Ting
論文名稱： 分子自組裝聚苯胺於幾丁聚醣凍凝膠對導電度增加及其在MC3T3-E1增生/分化的效應
The effect of self-doped polyaniline by molecular assembly in chitosan cryogel on the increasing conductivity and MC3T3-E1 proliferation
指導教授： 溫添進
Wen, Ten-Chin
學位類別： 碩士
Master
系所名稱： 工學院 - 化學工程學系
Department of Chemical Engineering
論文出版年： 2019
畢業學年度： 107
語文別： 中文
論文頁數： 76
中文關鍵詞： 冷凍凝膠 、自組裝聚苯胺 、組織工程
外文關鍵詞： cryogel, aza-michael addition, self-assembly poly aniline, tissue engineering
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    • 此研究的第一部分以不同比例的聚乙二醇二丙烯酸酯/幾丁聚醣(Poly(ethylene glycol) diacrylate/chitosan)作為前驅液，並且透過冷凍乾燥機，來形成組織工程的支架，不同比例的聚乙二醇二丙烯酸酯/幾丁聚醣冷凍凝膠分別名為Chitosan-PEGDA(0.5)、Chitosan-PEGDA(1)、Chitosan-PEGDA(2)、Chitosan-PEGDA(3)，其中後面的數字代表著加入聚乙二醇二丙烯酸酯的比例，數字越大代表加入的聚乙二醇二丙烯酸酯越多，用來分析以凍凝膠製程(cryogelation)開發大孔徑且具彈性的新型凍凝膠細胞支架。並且利用掃描式電子顯微鏡(scanning electron microscopy, SEM)可觀察到相連的孔洞結構，孔徑根據不同的比例分布於50-200 μm之間。再以FT-IR圖譜分析顯示兩種成份確定成功交聯於冷凍凝膠高分子壁中，最後再佐以MTT細胞毒性測驗以及萬用壓力測試儀所得到之數據來決定要以Chitosan-PEGDA(0.5)來繼續進行第二部分自組裝聚苯胺的實驗。
    在此研究的第二部分，則是以第一階段製造出來的冷凍凝膠為基礎，在高分子壁上自組裝聚苯胺來增加冷凍凝膠的導電度和機械強度。透過加入不同比例的苯胺以及鄰氨基苯磺酸之後以拉曼光譜來確定是否加入之苯胺以及鄰氨基苯磺酸有確實的自組裝於第一階段製備完的冷凍凝膠高分子壁上，之後再藉由萬用壓力測試儀發現機械強度相較於第一階段做出的冷凍凝膠增加了將近50倍，並且由於摻雜度的差異，導致在導電度上加入不同比例的苯胺以及鄰氨基苯磺酸，也會有所不同，造成自組裝之後的冷凍凝膠溝通電子的能力也有明顯的區別，最後再透過MTT細胞毒性測驗來選出對於培養細胞最具有優勢的冷凍凝膠，以作為之後觀察細胞分化使用的冷凍凝膠。未來可再經由加入生長因子以及提供不同的electrical stimulation來刺激細胞分化已培養出類似於天然骨的工程骨骼，進行骨骼缺陷的修復。

    The biodegradable shape memory polymers are candidate materials for making biomedical devices and scaffolds for tissue engineering. We had developed a biomimetic scaffold comprising chitosan and Poly(ethylene glycol) diacrylate. Therefore, we prepared macroporous chitosan/PEGDA cryogel scaffolds with elasticity via cryogelation process for cartilage tissue engineering.With different ratio of Poly(ethylene glycol) diacrylate/chitosan feeding in precursor solution, we set up four cryogel samples notation as Chitosan-PEGDA(0.5/1/2/3). The Chitosan-PEGDA(0.5/1/2/3) cryogels contained large, interconnected pores, open-cell strucure was confirmed by scanning electron microscopy (SEM) and Mercury Porosimeter. During stress-strain test for foam, Mechanical properties and porosity of the cryogels are closely related to the amount of PEGDA adding in the precursor solution. Mechanical property is highly positively related to the amount of PEGDA adding in the precursor solution but porosity is highly negatively related to the amount of PEGDA adding in the precursor solution. MTT assay showed that the Chitosan-PEGDA(0.5) has best noncytotoxic property. Therefore, we used Chitosan-PEGDA(0.5) as the template for anilin to self-dope on. As aniline o-aminobenzene sulfonic acid / was self-doped in the chitosan cryogel by molecular assembly, Chitosan-PEGDA(0.5) was coated with electrical nanofiber. With the self-doped polyaniline by molecular assembly in Chitosan-PEGDA(0.5), the conductivity is enhanced to 3×10^(-3) in hydrate state, 6.28×10^(-7) in dehydrate state. MTT assay showed that the self-doped polyaniline by molecular assembly in Chitosan-PEGDA(0.5) could extremely enhance the cell viability to 102.89%.

    摘要 I Extended Abstract III 致謝 XV 目錄 XVIII 表目錄 XX 圖目錄 XXI 第一章 緒論 1 1.1 前言 1 1.2 組織工程介紹 1 1.2.1 組織工程的基本概念 2 1.2.2 功能性細胞支架之設計 6 1.3 冷凍凝膠（Cryogel）技術 7 1.3.1 冷凍凝膠在組織工程上的技術發展 9 1.4 骨骼的組成 13 1.4.1 骨骼細胞的分類 15 1.4.2 骨骼的重塑 18 1.4.3 骨骼細胞的分化指標 20 1.4.4 造骨細胞生成之訊息傳遞路徑 22 1.4.5 骨骼細胞組織工程 25 1.5 材料介紹 29 1.5.1 幾丁聚醣 29 1.5.2 聚苯胺39 33 1.6 研究動機與目的 35 第二章 研究方法與步驟 37 2.1 實驗藥品 37 2.2 實驗流程 38 2.2.1 冷凍凝膠製程 38 2.2.2 聚苯胺分子間自組裝於冷凍凝膠表面 39 2.3 冷凍凝膠之分析 41 2.3.1 傅立葉轉換式紅外光譜儀（FT-IR）測試 41 2.3.2 水銀測孔儀（Mercury Porosimeter）測試 41 2.3.3 解析場發射掃描式電子顯微鏡分析（SEM） 41 2.3.4 電腦式萬能材料試驗機測試 42 2.3.5 膨潤比（Swelling Ratio）測定 43 2.3.6 細胞存活率分析（MTT assay by L929 cell） 44 2.4 聚苯胺分子間自組裝後之冷凍凝膠分析 45 2.4.1 拉曼光譜分析 45 2.4.2 導電度測試 46 2.4.3 解析場發射掃描式電子顯微鏡分析（SEM） 47 2.4.4 電腦式萬能材料試驗機測試 48 2.4.5 細胞存活率分析（MTT assay by L929 cell） 49 第三章 結果與討論 51 3.1 冷凍凝膠之材料分析 51 3.1.1 傅立葉轉換式紅外光譜儀（FT-IR）測試 51 3.1.2 冷凍凝膠結構分析 54 3.1.3 抗機械力分析 58 3.1.4 生物相容性測試 60 3.2 聚苯胺分子間自組裝冷凍凝膠之材料分析 62 3.2.1 拉曼光譜分析 62 3.2.2 聚苯胺分子間自組裝冷凍凝膠之導電度 63 3.2.3 解析場發射掃描式電子顯微鏡分析（SEM） 66 3.2.4 抗機械力分析 68 3.2.5 生物相容性測試 70 第四章 總結與建議 72 4.1 總結 72 4.2 未來之工作建議 73 第五章 參考資料 74

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