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研究生： 許筱芬
Hsu, Hsiao-Fen
論文名稱： 藉由空間侷限調控膠原蛋白與玻尿酸之層級式共組裝
Hierarchical Co-Assembly of Collagen with Hyaluronic Acid through Spatially-Confined Structures
指導教授： 李介仁
Li, Jie-Ren
學位類別： 碩士
Master
系所名稱： 理學院 - 化學系
Department of Chemistry
論文出版年： 2021
畢業學年度： 109
語文別： 中文
論文頁數： 99
中文關鍵詞： 細胞外間質共組裝 、空間侷限流體 、逐層沉積技術
外文關鍵詞： ECM co-assembly, spatially-confined, layer-by-layer
相關次數： 點閱：325  下載：0 
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    • 近年來，透過分子自組裝這樣類似天然形成的方式建構仿生材料已成為一個非常重要的議題。生物組織是由各種不同類型細胞精妙的集合體所組成，而細胞的外圍環境由非常複雜的細胞外間質所構成。細胞外間質提供了細胞的結構支撐、空間組織以及細胞與組織之間功能的整合。在細胞外間質中含量最豐富的蛋白質與醣分子分別為第一型膠原蛋白與玻尿酸，在人體內可相互共組裝形成方向性多層的層級式結構，但是要在體外重建此種細胞環境或是組織支架至今仍是一大挑戰。
    本研究將探討第一型膠原蛋白與玻尿酸的共組裝行為，玻尿酸的鏈狀結構會纏繞在第一型膠原蛋白奈米纖維的外圍，並且同時作為分隔物控制其排列。另外，玻尿酸的長度主要取決於其分子量，也會影響其與第一型膠原蛋白共組裝奈米纖維的型態。為了製作出有序排列的層級式第一型膠原蛋白與玻尿酸共組裝物，我們利用空間侷限結構控制其共組裝行為以及排列，並成功透過逐層沉積技術將第一型膠原蛋白與玻尿酸共組裝奈米纖維層層堆疊，製作出層級式多層仿生材料，說明了在體外模擬類細胞外間質的生物材料是具有可行性的。

    Biomimicry and the design of naturally inspired materials through self-assembly has now become an important concept in nanotechnology. Living tissues are intricate ensembles of multiple cell types embedded in a complex, well-defined ECM (extracellular matrix). ECM creates outer environment of cells, which provides structural support, spatial organization, and functional integration of cells and tissues. The most abundant ECM components are type I collagen (Col I) and hyaluronic acid (HA), which enable to co-assemble into highly ordered, layered and hierarchical structures. However, reconstructing such cell environment or tissue scaffold in vitro still remains a big challenge. In this study, we investigated the co-assembly of Col I and HA. The branches HA entwine with Col I nanofibrils and further serve as a spacer to adjust the alignment of Col I nanofibrils. Length of HA spacers depends on molecular weights, leading to different co-assembly behaviors of Col-HA nanofibrils. In order to artificially produce organized, hierarchical Col-HA co-assemblies, we utilized spatially-confined structures to control the co-assembly and alignment of Col I and HA. Through the layer-by-layer process, we successfully built up a biomimetic hierarchical multilayers composed of type I collagen nanofibrils entwined with HA. The results demonstrate that creating an ECM-like biomaterial is feasible.

    目錄 1 圖目錄 3 第一章 緒論 6 1.1 前言 6 1.2 細胞外間質(Extracellular Matrix, ECM) 6 1.2.1 第一型膠原蛋白 8 1.2.2 玻尿酸 10 1.3 蛋白質微奈米製程 11 第二章 探討第一型膠原蛋白與玻尿酸之共自組裝行為 16 2.1 前言 16 2.2 實驗部分 17 2.3 結果與討論 20 2.3.1 第一型膠原蛋白在雲母片上自組裝之型態 20 2.3.2 第一型膠原蛋白溶液培育時間對其在雲母片上自組裝的影響 20 2.3.3 玻尿酸在雲母片上自組裝之型態 21 2.3.4 第一型膠原蛋白與玻尿酸在雲母片共組裝之型態 21 2.3.5 第一型膠原蛋白與玻尿酸溶液培育時間對其在雲母片上共組裝的影響 22 2.3.6 玻尿酸濃度對第一型膠原蛋白與玻尿酸在雲母片上共組裝的影響 23 2.3.7 玻尿酸分子量對第一型膠原蛋白與玻尿酸在雲母片上共組裝的影響 25 2.3.8 離子強度對第一型膠原蛋白與玻尿酸共組裝的影響 29 2.3.9 培養溫度對第一型膠原蛋白與玻尿酸共組裝之影響 30 第三章 探討第一型膠原蛋白與玻尿酸在膠原蛋白奈米纖維結構上之共組裝行為 32 3.1 前言 32 3.2 實驗部分 33 3.3 結果與討論 37 3.3.1 玻尿酸分子量對第一型膠原蛋白與其在奈米纖維結構上共組裝的影響 37 3.3.2 固定劑對第一型膠原蛋白與玻尿酸在奈米纖維結構上共組裝的影響 38 3.3.3 玻尿酸分子量對第一型膠原蛋白與其在固定後奈米纖維結構上共組裝的影響 39 第四章 利用PDMS模板控制空間侷限流體內第一型膠原蛋白與玻尿酸共組裝形成多層結構 41 4.1 前言 41 4.2 實驗部分 42 4.3 結果與討論 47 4.3.1 不同空間侷限對第一型膠原蛋白與玻尿酸共組裝的影響 47 4.3.2 玻尿酸分子量對PDMS模板控制膠原蛋白與其共組裝的影響 49 4.3.3 玻尿酸分子量對奈米纖維結構引導膠原蛋白與其在空間侷限流體內共組裝的影響 51 4.3.4 固定劑對奈米纖維結構引導膠原蛋白與玻尿酸在空間侷限流體內共組裝的影響 53 4.3.5 空間侷限奈米纖維結構引導膠原蛋白與玻尿酸在空間侷限流體內共組裝 53 第五章 利用不同幾何結構的PDMS模板控制空間侷限流體內第一型膠原蛋白自組裝形成多層結構 56 5.1 前言 56 5.2 實驗部分 57 5.3 結果與討論 60 5.3.1 探討空間侷限流體內的第一型膠原蛋白奈米纖維於不同幾何結構上自組裝 60 5.3.2 探討不同的空間侷限流體內第一型膠原蛋白奈米纖維於相同幾何結構上自組裝 61 第六章 結論 64 第七章 未來方向：異種第一型膠原蛋白自組裝行為 67 7.1 實驗部分 67 7.2 結果與討論 69 7.2.1 探討(魚)第一型膠原蛋白在雲母片上自組裝之行為 69 參考文獻 71 圖附錄 75

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