單晶金屬有機骨架材料封裝活體細菌之研究

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李東(Dong Li) 查詢紙本館藏

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論文名稱

單晶金屬有機骨架材料封裝活體細菌之研究
(Bacterial Microrobot for the Encapsulation of Living Bacteria in a Single Metal-Organic Frameworks)

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至系統瀏覽論文 (2028-6-30以後開放)

摘要(中)

癌症為全球死因第二位，每年約有一千萬人死於癌症。而在台灣，癌症已連續四十年蟬聯十大死因的首位。且癌症發生率也年年上升。癌症的傳統療法常具有同時傷害癌症細胞與一般細胞的性質，身體虛弱的癌症病患不僅需承受強大的副作用，且治療過程往往十分漫長。因此需要開發新型且有效的癌症治療方法。在醫學上，有使用細菌進行免疫反應之免疫療法。免疫療法是藉由活化人體的免疫系統，來達到殺傷癌細胞的效果。但癌症病患的體內常有抗生素與溶菌酶等其他療法所施打的藥物，導致微生物降低活性甚至死亡。因此科學家便致力研究結合微生物的生物複合材料，來增強生物細胞對環境的適應性。
本研究利用本實驗室於2015年發表於JACS之文章，利用類沸石咪唑骨架材料-90(ZIF-90)封裝酵素之技術，合成微米級ZIF-90封裝大腸桿菌(E .coli@ZIF-90)，並與先前發表之多晶包覆大腸桿菌奈米級類沸石咪唑骨架材料-8(E. coli⸦ZIF-8)比較。發現經由紫外光滅菌程序，仍能保持單晶包覆之E . coli@ZIF-90內大腸桿菌約百分之八十的存活率。且在照射紫外光時間增加後，大腸桿菌之存活率沒有明顯下降。並合成封裝抗癌藥物之奈米級ZIF-90，期望將來可將抗癌藥物協同細菌癌症療法。以開創嶄新的癌症療法。

摘要(英)

Cancer has remained the leading cause of death in Taiwan, with rising incidence rates annually. Conventional cancer treatments pose risks to both cancer cells and healthy cells, resulting in severe side effects and lengthy procedures for weakened patients. Microbial-mediated treatments provide a targeted approach by selectively colonizing tumors, inhibiting cancerous growth, and minimizing negative impacts compared to less precise methods. However, concerns arise regarding the potential spread, mutation, and acquisition of antibiotic resistance by attenuated bacteria. Enhancing targeting efficiency and identifying safe and effective bacterial strains continue to pose challenges in this field.
According to our previous report, in this study, our laboratory successfully synthesized micron-scale Zeolitic imidazolate frameworks-90 (ZIF-90)-encapsulated E. coli (E. coli@ZIF-90). To make comparisons, we examined the poly-nanocrystalline encapsulated E. coli known as Zeolitic imidazolate frameworks -8 (E. coli⸦ZIF-8). Despite increased exposure to UV light, the single-crystal coated E. coli@ZIF-90 displayed an impressive 80% survival rate of encapsulated bacteria, showcasing exceptional preservation during UV sterilization with no significant decline in E. coli′s survival rate. Furthermore, our research successfully synthesized nanoscale ZIF-90 for the encapsulation of anti-cancer drugs, demonstrating the potential for synergistic approaches that integrate anti-cancer drugs and bacterial cancer therapy, paving the way for a novel paradigm in cancer treatment with enhanced therapeutic outcomes.

關鍵字(中)

★ 金屬有機框架
★ 微生物介導療法
★ Zeta 電位
★ 大腸桿菌
★ 植物乳桿菌B21

關鍵字(英)

★ Metal-organic Frameworks
★ Microbial-Mediated Therapy
★ Zeta Potential
★ Escherichia coli
★ Lactiplantibacillus plantarum B21

論文目次

摘要 i
Abstract ii
圖目錄 viii
表目錄 viii
1 第一章緒論 1
1-1 金屬有機骨架材料 1
1-1-1 金屬有機骨架材料 1
1-1-2 類沸石咪唑骨架材料 3
1-1-3 類沸石咪唑骨架材料-8/-90 5
1-1-4 鋁基金屬有機骨架材料A520 5
1-2 微生物 6
1-2-1 微生物 6
1-2-2 大腸桿菌(Escherichia coli, E .coli) 6
1-2-3 植物乳桿菌B21(Lactiplantibacillus plantarum B21, L. plantarum B21) 7
1-2-4 質體(Plasmid) 8
1-2-5 癌症療法 10
1-2-6 細菌癌症療法(Bacterial Cancer Therapy) 11
1-3 研究動機與目的 14
2 第二章實驗 16
2-1 實驗藥品與設備 16
2-1-1 實驗藥品 16
2-1-2 實驗儀器 19
2-1-3 場發掃描式電子顯微鏡(Field-emission Scanning Electron Microscope, FE-SEM) 21
2-1-4 X射線粉末繞射儀(Power X-ray Diffractometer, PXRD) 22
2-1-5 攜帶式分光光度計(Ultrospec 10 cell Density Meter) 24
2-1-6 紫外光可見光分光光譜儀(UV/VIS Spectrophotomter) 24
2-1-7 界面電位分析儀(Zeta Potential Analyzer) 24
2-2 實驗使用之酵素 26
2-2-1 溶菌酶(Lysozyme) 26
2-2-2 蛋白酶 (Protease) 26
2-3 實驗步驟 27
2-3-1 大腸桿菌之培養步驟 27
2-3-2 微米級類沸石咪唑骨架材料-90封裝之大腸桿菌(E .coli@ZIF-90)之合成步驟 28
2-3-3 奈米級類沸石咪唑骨架材料-8塗層之大腸桿菌(E .coli⸦ZIF-8)之合成步驟 29
2-3-4 微米級類沸石咪唑骨架材料-90封裝之大腸桿菌(E .coli@ZIF-90)抵抗紫外光能力測試實驗步驟 29
2-3-5 奈米級類沸石咪唑骨架材料-8塗層之大腸桿菌(E .coli⸦ZIF-8)抵抗紫外光能力測試實驗步驟 30
2-3-6 大腸桿菌(E .coli)抵抗紫外光能力測試實驗步驟 30
2-3-7 奈米級類沸石咪唑骨架材料-90合成步驟 31
2-3-8 單批微米級類沸石咪唑骨架材料-90封裝之大腸桿菌(E. coli@ZIF-90)含有之活體 E. coli 數量測定 31
2-3-9 奈米級類沸石咪唑骨架材料-90包覆阿黴素(DOX@nano ZIF-90)之合成步驟 32
2-3-10 奈米級類沸石咪唑骨架材料-90包覆腎上腺皮質酮(Prednisolone@nano ZIF-90)之合成步驟 32
2-3-11 奈米級類沸石咪唑骨架材料-90塗層之大腸桿菌(E .coli⸦ nano ZIF-90)之合成步驟 33
2-3-12 奈米級類沸石咪唑骨架材料-90塗層之大腸桿菌再次結晶實驗之相關步驟 33
2-3-13 奈米級類沸石咪唑骨架材料-8塗層之大腸桿菌(E. coli⸦ZIF-8)界面電位測定步驟 34
2-3-14 植物乳桿菌B21之培養步驟 34
2-3-15 鋁基金屬有機骨架材料A520塗層之植物乳桿菌B21之合成步驟 35
3 第三章結果與討論 36
3-1 大腸桿菌生物複合材料之相關鑑定 36
3-1-1 X射線粉末繞射儀之鑑定結果 36
3-1-2 場發掃描式電子顯微鏡之成像結果 37
3-1-3 單批微米級類沸石咪唑骨架材料-90封裝之大腸桿菌(E. coli@ZIF-90)含有之活體 E. coli 數量測定 38
3-1-4 微米級類沸石咪唑骨架材料-90 /奈米級類沸石咪唑骨架材料-8塗層之大腸桿菌(E .coli@ZIF-90/E .coli⸦ZIF-8)抵抗紫外光能力測試 39
3-2 抗癌藥物生物複合材料之相關鑑定 42
3-2-1 X射線粉末繞射儀之鑑定結果 42
3-2-2 場發掃描式電子顯微鏡之成像結果 44
3-2-3 奈米級類沸石咪唑骨架材料-90包覆阿黴素測試 46
3-2-4 奈米級類沸石咪唑骨架材料-90包覆腎上腺皮質酮測試 48
3-2-5 界面電位對於類咪唑骨架材料-8結晶大小之影響測試 49
3-3 植物乳桿菌B21生物複合材料之相關鑑定 51
3-3-1 X射線粉末繞射儀之鑑定結果 51
3-3-2 場發掃描式電子顯微鏡之成像結果 52
3-3-3 單批奈米級A520塗層之L. plantarum B21 (B21⸦A520)含有之活體 L. plantarum B21數量測定 53
4 第四章結論與未來展望 55
參考文獻 56

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指導教授

謝發坤

審核日期

2023-8-14

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