臺灣博碩士論文加值系統

肺炎，近年來已成為高罹患率與高死亡率的疾病。健大黴素(Gentamicin, GM) 為胺基醣苷類 (Aminoglycoside) 抗生素之一，常被用來治療肺炎等疾病。不過，由於其口服吸收率低，長期使用又會導致腎毒性 (Nephrotoxicity) 及耳毒性 (Ototoxicity) 副作用，使得此藥物使用上面臨重大限制。本研究製備幾丁聚醣/褐藻醣膠奈米粒攜帶健大黴素，探討經肺部傳輸系統對抗肺炎之研究。結果顯示，幾丁聚醣/褐藻醣膠奈米粒可經由正負電荷交互作用形成，當幾丁聚醣與褐藻醣膠的重量比為5:1時，於模擬肺液中粒徑分佈約246.1 nm ；以此奈米粒包覆健大黴素後，於模擬肺液中粒徑分佈約271.1 nm，包覆率高達94%，控制釋放百分比累計72 hr達99%。細胞毒性結果顯示，對 Raw 264.7 cell 而言，奈米粒之 IC50 約為 0.8 mg/mL，對A549 與 Raw 264.7具有良好的生物相容性；此奈米粒不會引起細胞發炎反應；具有降 DPPH 自由基以及LPS 所誘導之 ROS之功效；抑菌實驗顯示，以此奈米粒包覆健大黴素後可有效降低克雷伯氏肺炎桿菌 (Klebsiella pneumonia) 之最低抑菌濃度 (minimum inhibitory concentrations, MIC) 至 1.95 μg/mL 。由以上得知，幾丁聚醣/褐藻醣膠奈米粒可經肺部傳輸系統控制釋放健大黴素達對抗肺炎之功效。

Pneumonia is one of the leading causes of morbidity and mortality. Gentamicin (GM) is an aminoglycoside antibiotic commonly used for treating pneumonia disease. However, the use of GM is limited by a relatively short half-life and the dose-related adverse effects as ototoxicity and nephrotoxity is reflected in its low therapeutic index. In this investigation, we devel¬oped a controlled-release GM formulation using chitosan/fucoidan nanoparticles (CS/F NPs). The results suggest that nanoparticles composed of CS and F were successfully prepared based on their electrostatic interaction. The mean particle size of CS/F NP was around 246.1 nm as the weight ratio of CS to F was 5:1 and GM loaded CS/F NPs (GM/CS/F NPs) was around 271.1 nm in simulated lung fluid. The GM can be highly encapsulated and sustained release from CS/F NPs for up to 72 hrs. The MTT cell viability assay revealed an absence of overt toxicity to A549 or Raw 264.7 cells following exposure to the formulations containing < 0.8 mg NP/ml for up to 24 hrs. Inflammation and intracellular ROS assay revealed that CS/F NPs are suitable biomaterials without inducing inflammatory or oxidative reactions. GM-loaded CS/F NPs are effective in resisting Klebsiella pneumonia with low minimum inhibitory concentrations (MICs) at 1.95 μg/mL. Briefly, all the results mentioned above suggest that the CS/F NPs developed in this study are a promising carrier for pulmonary delivery of GM against pneumonia.

目錄
謝誌......................................I
摘要................ II
Abstract …………………………………………………………………………….III
目錄................ IV
圖目錄.......... VI
表目錄...... VIII
第一章、前言 1
第二章、文獻回顧 2
2-1、肺炎（PNEUMONIA） 2
2-2、GENTAMICIN (健大黴素，GM) 3
2-2-1、Gentamicin基本介紹 3
2-2-2、Gentamicin 的抗菌機制 3
2-2-3、Gentamicin 的毒性機制與保護 4
2-3、奈米技術 4
2-3-1、奈米粒載體 4
2-3-2、奈米載體控制釋放 5
2-3-3、奈米載體傳遞路徑 5
2-4、肺部傳遞 (PULMONARY DELIVERY) 5
2-4-1、呼吸系統的組成 5
2-4-2、呼吸道的結構與特性 6
2-4-3、氣霧粒子沉積機制 7
2-4-4、黏膜纖毛清除機制 7
2-4-5、肺泡巨噬細胞清除機制 7
2-4-6、肺部傳遞常用設備 8
2-5、幾丁聚醣 (CHITOSAN,CS) 9
2-5-1、幾丁聚醣結構 9
2-5-2、幾丁聚醣生理活性及生醫材料上的應用 9
2-6、褐藻醣膠 (FUCOIDAN,F) 10
2-6-1、褐藻醣膠結構 10
2-6-2、褐藻醣膠生理活性 10
2-6-3、褐藻醣膠生醫材料上的應用 11
2-7、克雷伯氏肺炎桿菌 (KLEBSIELLA PNEUMONIA, K.P.) 12
第三章、實驗動機與目的 13
第四章、實驗架構 14
第五章、實驗材料與方法 15
5-1、實驗材料 15
5-2、實驗儀器 17
5-3、實驗方法 18
5-3-1、幾丁聚醣/褐藻醣膠奈米粒之特性實驗 18
5-3-2、幾丁聚醣/褐藻醣膠/Gentamicin奈米粒之特性實驗 19
5-3-3、體外細胞實驗 21
5-3-4、動物實驗 26
第六章、結果與討論 28
6-1、幾丁聚醣/褐藻醣膠奈米粒之特性實驗 28
6-2、幾丁聚醣/褐藻醣膠/Gentamicin奈米粒之特性實驗 29
6-3、幾丁聚醣/褐藻醣膠/Gentamicin奈米粒之體外實驗 31
6-4、Gentamicin 於動物體內之生物可利用率 35
第七章、結論 36
第八章、未來展望 37
第九章、參考文獻 38
第十章、圖表 45

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