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dc.contributor.authorChang, Wen-Hsinen_US
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dc.description.abstract明膠為具有良好生物可降解性、生物相容性之天然高分子,則廣泛運用於製備生醫材料支架方面,但由於機械性質、熱穩定性不佳與降解速率過快的因素,本研究將複合不同濃度綠槴子素、戊二醛、EDC/NHS、PBAAM及綠槴子素合併使用與明膠進行交聯處理,使明膠具有較穩固的結構並補強純明膠材料的機械性質和抗降解性,再利用冷凍乾燥法製備出具多孔性三維之明膠支架,以成為理想的生物植入支架。研究中並同時探討交聯劑種類和濃度對支架的物理性質、結構穩定性以及體外生物相容性之影響,評估此明膠支架是否利於組織之再生及修復,進而應用於生醫領域上。 UV-Vis可觀察交聯明膠支架之交聯指數,添加不同濃度之綠槴子素、戊二醛、EDC/NHS、PBAAM/綠槴子素與明膠交聯處理之支架交聯程度分別約為91%、25~81%、64~83%、86~90%。FT-IR結果顯示交聯明膠支架皆有著生物高分子蛋白質所擁有的醯胺基之特性峰。以S.E.M觀察明膠支架具有良好相互連結的多孔結構,孔徑範圍為200~520μm間的三維空間結構,具有符合細胞向內生長的環境;吸水率會隨著明膠支架之交聯程度較高,結構較穩固,而吸水率會減低,且製備出明膠支架的孔隙率介於77~92%間。機械性質檢測結果顯示添加交聯劑確實能夠有效提升純明膠支架之機械強度,尤以綠槴子素及PBAAM/綠槴子素交聯處理明膠支架的機械強度增強34~89倍。 在體外生物降解測試中,經由交聯鍵結作用可以提升支架結構的穩定性,有效減緩純明膠的降解速率,從重量損失結果可證實PBAAM/綠槴子素交聯明膠支架中較符合多孔性支架對於細胞培養,是生物可降解與可提供足夠空間與時間待細胞生長和組織形成。在生物相容性測試中,植入鼠的纖維母細胞(L929)至支架共培養後,並經由螢光顯微鏡與S.E.M結果顯示,以合併使用PBAAM與綠槴子素交聯明膠支架有利於細胞正常貼附與生長,具有良好的生物相容性。 綜合上述結果,本研究中以PBAAM/綠槴子素交聯明膠支架中具有較佳交聯指數、適當孔徑大小、吸水率、理想孔隙率、機械強度提升、適當的生物降解速率以及合適細胞貼附增生的環境與良好的生物相容性,研究中製備出明膠支架利於組織之再生及修復,可作為理想組織工程支架植入材,進而應用於生醫領域上。zh_TW
dc.description.abstractGelatin, a natural polymer, shows high biocompatibility and biodegradability and can be widely used in biomedical application. In this study, the porous and three-dimensional gelatin scaffolds with various concentration of crosslinking agents, such as genipin, glutaraldehyde, EDC/NHS and PBAAM/genipin, were successfully fabricated using freeze-drying technique. The results of UV-Visible spectrophotometer (UV-Vis) show the crosslinking index of gelatin scaffold of genipin, glutaraldehyde, EDC/NHS and PBAAM/genipin are in the range of 91%, 25~81%, 64~83%, 86~90%, respectively, depending on their concentration. The data of Fourier Transform Infrared spectrophotometer (FT-IR) of crosslinked gelatin scaffold represent the amide group of characteristic peaks of biopolymer proteins. The images of scanning electron microscope (SEM) of gelatin scaffold reveal the well interconnected porous structure, and the pore size is in the range of 200~520μm between three-dimensional structure for cell ingrown environment. Water absorption of gelatin scaffold is related to crosslinking index. While crosslinking index is increased, the structure is more stable, leading to the lower water absorption. The porosity of gelatin scaffold is between 77~92%. The mechanical properties of gelatin scaffold with genipin and PBAAM/genipin could be significantly enhanced 34~89 times in magnitide compared to the pure gelatin scaffold. For the in vitro biodegradation test, the stability of gelatin structure could be enhanced by the crosslinking process and the degradation rate of gelatin scaffold decreased by adding crosslinking agents. The weight loss of gelatin scaffold with PBAAM/genipin confirms that biodegradable porous structure could provide enough space to be cell growth and tissue formation. In biocompatible test, mouse fibroblasts cell (L929) were seeded into gelatin scaffold. The images of fluorescent microscopy and SEM show that gelatin scaffold with PBAAM/genipin is biocompatible and favorable for cell attachment and growth. The gelatin scaffold with PBAAM/genipin containing better crosslinking index, appropriate pore size, water adsorption and porosity, suitable mechanical strength and biodegradable rate can prodive favorable environment of cell adhesion, proliferation, and well biocompatability. The physical properties of fabricated gelatin scaffolds indicate that the addition of crosslinking agent can improve their biodegradable structure. Preparation of gelatin scaffold applied in the organization of the regeneration/repair and tissue engineering could be used in the applications of implanted material and biomedical field.en_US
dc.description.tableofcontents摘要 I Abstract II 總目錄 IV 圖目錄 VI 表目錄 IX 第一章 緒論 1 1-1 研究背景 1 1-2 研究動機 3 1-3 研究方向 3 第二章 文獻回顧 4 2-1 組織工程 4 2-2 生醫材料 6 2-2-1 合成高分子材料 6 2-2-2 生物高分子材料 7 2-3 高分子生醫材料之製備方法 10 2-3-1 鹽析法 11 2-3-2 氣體成形法 11 2-3-3 相分離法 11 2-3-4 纖維鍵結法 12 2-3-5 冷凍乾燥法 12 2-4 明膠之交聯機制 19 2-4-1 綠槴子素 20 2-4-2 戊二醛 25 2-4-3 EDC / NHS 27 2-5 高分子之降解機制 30 2-5-1 高分子之水解機制 31 2-5-2 高分子之酶解機制 32 2-5-3 明膠生物降解之特性 32 2-6 細胞機制 34 2-6-1 細胞週期 34 2-6-2 生醫支架與細胞貼附 35 第三章 實驗方法與步驟 39 3-1 實驗材料 39 3-2 實驗儀器 41 3-3 實驗流程圖 42 3-4 實驗步驟 43 3-4-1 明膠複合支架之製備 43 3-4-2 明膠複合支架之交聯指數分析 44 3-4-3 明膠複合支架之生物降解 45 3-4-4 明膠複合支架之細胞貼附實驗 46 3-5 明膠複合支架之材料性質分析 48 3-6 實驗儀器分析 49 3-6-1 掃描式電子顯微鏡(S.E.M) 49 3-6-2 傅立葉轉換紅外線光譜儀(FT-IR) 49 3-6-3 紫外光-可見光分析光譜儀(UV-Vis) 49 3-6-4 萬能試驗機 49 3-6-5 倒立式顯微鏡 50 3-6-6 螢光顯微鏡 50 3-7 實驗代碼 51 第四章 結果與討論 52 4-1 交聯明膠支架之材料特性分析 52 4-1-1 交聯程度測試 53 4-1-2 交聯明膠支架之結構鑑定 56 4-1-3 明膠支架之巨觀型態 59 4-1-4 明膠支架之微觀孔洞型態 60 4-1-5 明膠支架之吸水率測試 65 4-1-6 明膠支架之孔隙率測試 67 4-1-7 明膠支架之機械性質檢測 69 4-2 交聯明膠支架之生物降解測試 73 4-3 交聯明膠支架之生物相容性測試 79 第五章 結論 87 參考文獻 89zh_TW
dc.subjectcrosslinking agenten_US
dc.titlePreparation and characterization of porous three-dimensional gelatin scaffold using different crosslinking agentsen_US
dc.typeThesis and Dissertationzh_TW
item.openairetypeThesis and Dissertation-
item.fulltextno fulltext-
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