臺灣博碩士論文加值系統

本研究使用四種不同碳鏈長的醛類及N-(2-羥基)丙基-3-甲基二丙烯基四級銨鹽 glycidyl-methyl-diallyl amonium salt (GMDAS)修飾幾丁聚醣，合成得到四種不同碳鏈長的新型雙陽離子之N-O-四級銨鹽化幾丁聚醣衍生物(DCQCS)，其目的是為了強化幾丁聚醣的抗菌性及生物相容性。所得之四種幾丁聚醣衍生物DCQCS-MD、DCQCS-ED、DCQCS-PD及DCQCS-BD，以傅立葉轉換紅外光光譜儀（FT-IR）分析其分子結構，X光繞射檢視結晶結構，介達電位（Zeta potential）分析幾丁聚醣及衍生物之電位性質，以熱重分析儀（TGA）及示差掃描量熱分析（DSC）測試熱裂解溫度、熱穩定性及熱性質變化，共軛焦顯微鏡觀察幾丁聚醣衍生物探討與細胞膜的融合與穿透性，並以金黃色葡萄球菌（Staphylococcus aureus）及肺炎桿菌（Klebsiella pneumonia）測試最小殺菌濃度（MBC）。另外，以老鼠纖維母細胞（L929）測試其細胞成長活性與細胞毒性，期望未來能應用於新型藥物釋放載體、傷口敷材及其他生醫領域。

In this study, chitosan was grafted with glycidyl-methyl-diallyl ammonium salts (GMDAS) to form new N-O-dicationic quaternary ammonium salts chitosan derivatives (DCQCS) with four carbon chain lengths. The purpose is to strengthen the chitosan antimicrobial resistance and biocompatibility. These four DCQCS were examined using Fourier transform infrared spectroscopy (FTIR) to determine the chemical structures, X-ray diffraction (XRD) to determine their crystal structure, Zeta potential to determine their surface charges, TGA and DSC to measure their pyrolysis temperature, thermal stability and thermal properties. The fusion and uptake of DCQCS by fibroblasts was observed by Confocal microscopy. Antibacterial activity was evaluated against Staphylococus aureus and Klebsiella pneumonia based on the minimal bactericidal concentration. The biocompatibility and cytotoxicity were evaluated with mouse fibroblasts (L-929). The results show that DCQCS can be applied as drug delivery carriers, wound dressing materials, and other biomedical fields in the future.

目錄
摘要 I
Abstract II
目錄 III
圖目錄 VI
表目錄 X
第一章 緒論 1
1.1 研究背景 1
1.2 研究目的 2
第二章 文獻回顧 3
2.1 幾丁質與幾丁聚醣簡介 3
2.2 幾丁質與幾丁聚醣的應用 5
2.3 幾丁聚醣抗菌機制 8
2.4 修飾幾丁聚醣 9
2.5 四級銨鹽 10
2.6 微生物 11
2.6.1 細菌之生長 12
2.6.2 影響細菌生長因素 13
2.6.3 金黃色葡萄球菌 14
2.6.4 肺炎桿菌 15
2.7 細胞介紹 15
2.7.1 細胞結構 15
2.7.2 細胞生長週期 16
2.7.3 細胞衰老與細胞凋亡 17
第三章 實驗步驟 18
3.1 實驗藥品 18
3.2 實驗儀器 20
3.3 實驗流程 21
3.4 樣品製備 22
3.4.1 製備四級銨鹽化合物 22
3.4.2 製備雙陽離子之N-O-四級銨鹽化幾丁聚醣衍生物 23
3.5 基本性質測試 24
3.5.1 傅立葉紅外線光譜分析（FT-IR） 24
3.5.2 X光繞射 24
3.5.3 熱重分析(TGA) 24
3.5.4 示差掃描量熱分析 25
3.5.5 Zeta potential 分析 25
3.6 抗菌實驗 25
3.6.1 菌活性質之計算 25
3.6.2 抗菌活性 26
3.7 細胞相容性測試 26
3.7.1 細胞活化 26
3.7.2 細胞繼代 27
3.7.3 纖維母細胞於幾丁聚醣衍生物溶液中培養 27
3.7.4 細胞增生實驗（MTT assay） 27
3.8 共軛焦顯微鏡(Confocal Microscopy) 28
第四章 結果與討論 29
4.1 傅立葉紅外線光譜分析(FT-IR) 29
4.2 X光繞射 30
4.3 熱重分析(TGA) 31
4.4 示差掃描量熱分析 34
4.5 Zeta Potential測定 35
4.6 抗菌實驗 36
4.7 細胞相容性測試 38
4.7.1 細胞型態的觀察 38
4.7.2 細胞增生實驗(MTT assay) 46
4.8 共軛焦顯微鏡(Confocal Microscopy) 51
第五章 結論 62
第六章 參考文獻 64

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