羥基自由基(•OH)被認為是强力的活性氧化物質(ROS)，其抗菌活性超過其他種類的ROS。•OH可由芬頓(Fenton)反應所產生，該反應需要過氧化氫（H2O2）做為反應物，亞鐵離子（鐵（II））為催化劑。在本論文中使用含有葡萄糖氧化酶(GOx)的奈米反應器，通過GOx與葡萄糖(Glu)反應產生H2O2，再由奈米反應器中所含之二價鐵化合物(Hem)或所加入亞鐵離子來促使芬顿反應發生產生羥基自由基(•OH)。論文中採用了兩種不同的奈米反應器結構，即PVA電紡奈米纖維和沸石咪唑骨架(ZIF-8)奈米粒子。在製備過程中將含有葡萄糖的PVA溶液與含有GOx的PVA溶液同時進行電紡來製造新型的奈米纖維反應器。含有葡萄糖的奈米纖維主要是為了提供在分隔的GOx奈米纖維葡萄糖以產生H2O2。而對於ZIF-8奈米反應器而言，則需在反應過程中額外添加葡萄糖。以TMB為反應基質可證實所製備的反應系統具有類過氧化物酶的活性，在可見光光譜分析中可以觀測到在~450nm或~650nm處顯示出明顯的峰值；使用對苯二甲酸(TPA) 為基質可證明反應系統中•OH的產生，當TPA與•OH反應後•OH生成螢光羥基-對苯二甲酸(HTPA)，可以在PL發射光譜觀測到波長~425nm處之峰值。最後分別以革蘭氏陽性菌（金黄色葡萄球菌）和革蘭氏陰性菌（大腸杆菌）針對各系统進行抗菌活性實驗，與H2O2的抗菌活性相比，此兩系統所產生的•OH有明顯更強的抗菌效果。所有菌體在濃度OD值為1時與奈米系統接觸12小時後均能達到100%的滅殺率。

Developing antimicrobial materials with high effectiveness while maintaining their biocompatibility and biodegradability has been becoming more important recently, especially ever since the starting of pandemic. Hydroxyl radical (•OH) has been known as the most powerful reactive oxygen species (ROS) with antimicrobial activity surpassing any of other ROS. The generation of •OH comes from Fenton reaction, which requires hydrogen peroxide (H2O2) as a reactant and ferrous ion (iron (II)) as a catalyst. In this study, nanosystems containing glucose oxidase (GOx), which produces H2O2 through reaction with glucose (Glu), with iron ion containing hemin (Hem) or direct ferrous ion were employed to generate •OH by supplementing the Fenton reaction. Two different nanosystems were adopted, which are PVA electrospun nanofibers and zeolitic imidazole framework (ZIF-8) nanoparticles, to ensure the biocompatibility of these materials. Nanofibrous reactor was fabricated by employing co-electrospinning of two separated Glu and GOx containing PVA dope solutions. Glu nanofibers is used to supply the required substrate of GOx encapsulated in the separated nanofibers for H2O2 generation. While for ZIF-8 nanosystems, supplementary Glu was added throughout the reaction. Characterizations of each nanosystem were done using FE-SEM, FTIR, and XRD. The peroxidase-like activity of as-prepared nanosystem was confirmed through TMB reaction with peaks showing at either ~450nm or ~650nm, while generation of •OH was detected under terephthalic acid (TPA) reaction to generate fluorescent hydroxyl-terephthalic acid (HTPA) measured using PL emission spectroscopy at emission wavelength ~425nm. Antimicrobial activity of each nanosystems were tested against both Gram-positive (S. auerus) and Gram-negative bacteria (E. coli), which shown much higher antimicrobial results as compared to the antimicrobial activity of H2O2. 100% killing rate was achieved for all nanosystems at 12h contact time against OD=1 for both bacteria under 0.2%(w/v) of nanofibers and 0.05% (w/v) of ZIF-8 nanoparticles.

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