臺灣博碩士論文加值系統

由於傳統抗生素的過度使用，導致細菌出現耐藥性，並造成全球性的健康威脅。因此，研發可替代傳統抗生素的抗菌材料，便成為近年迫在眉睫之研究重點。碳量子點（CQDs）為一種新型碳基奈米材料，因其具備微小尺寸（< 10 nm）、優異的生物相容性及獨特的光學特性，使其在對抗耐藥性感染方面展現強大的潛力。本研究將不可食用的石榴皮（pomegranate peel）分別透過燒結法（pyrolysis）及水熱法（hydrothermal），將以往視為廢棄物的石榴皮製成具有高價值的石榴皮碳量子點（ppCQD）。本研究除了分析不同合成方法之材料特性，亦結合藍光（440 nm）照射，激發ppCQD。並進一步評估ppCQD對於金黃色葡萄球菌（Staphylococcus aureus）之光動力抗菌效益。實驗結果顯示，透過燒結及水熱合成法製備之碳量子點在無照光條件下培養4小時，濃度為400 μg mL-1的ppCQD對金黃色葡萄球菌之殺菌效率便可達50 %。此外，在照射4小時藍光後，其對金黃色葡萄球菌的殺菌效率更可達近80 %。這些數據表明，使用石榴皮碳量子點進行抗菌光動力療法殺滅金黃色葡萄球菌是一種具發展性且可行的方式。

The excessive use of antibiotics causes the emergence of bacterial resistance, which has become a global health threat. Therefore, it has become urgently important to develop alternative antimicrobial materials. Carbon quantum dots（CQDs）, a new type of carbon-based nanomaterials, shows great potential against antimicrobial-resistant infections due to their favorable characteristics, including their tiny size（< 10 nm）, excellent biocompatibility and unique optical properties. In this study, the inedible pomegranate peel was transformed into valuable pomegranate peel carbon quantum dots（ppCQDs）using two different synthesis methods : pyrolysis and hydrothermal synthesis. Alongside the characterization of the materials produced using each of these methods, the photodynamic antibacterial efficacy of ppCQDs against Staphylococcus aureus was evaluated under blue light（440 nm）excitation. For instance, after four hours of culture without illumination, hydrothermally produced ppCQDs at a concentration of 400 μg mL-1 showed 50 % bactericidal efficacy against Staphylococcus aureus. However, the bactericidal effectiveness against Staphylococcus aureus could be increased to 80 % after 4 hours of blue light irradiation. According to the data, using ppCQDs for antimicrobial photodynamic treatment is a viable and promising strategy for eliminating Staphylococcus aureus.

摘要 ..... i
Abstract ..... ii
誌謝 ..... iii
目錄 ..... iv
表目錄 ..... vi
圖目錄 ..... vii
縮寫對照表 ..... viii
第一章 緒論 ..... 1
1.1 前言 ..... 1
1.2 研究目的 ..... 1
第二章 文獻探討 ..... 2
2.1 奈米碳材料 ..... 2
2.1.1 碳量子點（CQDs） ..... 2
2.2 碳量子點的合成 ..... 2
2.2.1 自上而下合成 ..... 3
2.2.2 自下而上合成 ..... 3
2.3 碳量子點的應用 ..... 4
2.3.1 生物成像 ..... 4
2.3.2 離子檢測 ..... 5
2.3.3 藥物傳輸 ..... 5
2.3.4 抗菌 ..... 6
2.4 碳量子點的抗菌途徑 ..... 6
2.4.1 尺寸的影響 ..... 6
2.4.2 表面電荷 ..... 6
2.4.3 光動力抗菌 ..... 7
2.4.4 官能基 ..... 7
第三章 實驗架構與材料方法 ..... 8
3.1 實驗架構圖 ..... 8
3.2 實驗材料 ..... 9
3.2.1 實驗藥品 ..... 9
3.2.2 實驗儀器 ..... 11
3.2.3 實驗菌株 ..... 12
3.2.4 實驗細胞 ..... 12
3.3 實驗方法 ..... 13
3.3.1 碳量子點製備及分析 ..... 13
3.3.1.1 碳量子點水熱合成 ..... 13
3.3.1.2 碳量子點高溫燒結 ..... 13
3.3.1.3 場發射式穿透顯微鏡（FE-TEM） ..... 13
3.3.1.4 動態光散射粒徑分析儀（DLS） ..... 13
3.3.1.5 介面電位分析（Zeta potential） ..... 14
3.3.1.6 傅立葉紅外光譜（FTIR） ..... 14
3.3.2 細胞培養及繼代 ..... 14
3.3.3 細胞存活率測試（MTT） ..... 14
3.3.4 菌株活化 ..... 15
3.3.5 生長曲線及稀釋濃度測定 ..... 15
3.3.6 抗菌能力測定(OD 600) ..... 15
3.3.7 抗菌能力測定(平版計數) ..... 15
3.3.8 活性氧物質(ROS) ..... 16
3.3.9 熱場發射掃描式電子顯微鏡（FE-SEM）之菌體型態分析 ..... 16
3.3.10 統計分析 ..... 16
第四章 結果與討論 ..... 17
4.1 水熱/燒結法PPCQD之物化分析 ..... 17
4.1.1 場發射式電子穿透顯微鏡（FE-TEM） ..... 17
4.1.2 場奈米粒徑及介面電位測量 （DLS & zeta potential） ..... 18
4.1.3 傅立葉轉換紅外光譜 （FTIR） ..... 19
4.2 體外細胞實驗 （HEK-293） ..... 21
4.2.1 細胞存活率（MTT） ..... 21
4.3 抗菌測試（金黃色葡萄球菌） ..... 21
4.3.1 抗菌效率測試（OD 600） ..... 21
4.3.2 抗菌效率測試（塗盤法） ..... 22
4.3.3 活性氧化物試驗（ROS） ..... 23
4.3.4 菌體分析（FE-SEM） ..... 24
第五章 結論 ..... 25
參考文獻 ..... 27
Extended Abstract ..... 33

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