藥物的濫用，常常會造成抗藥性細菌的生成，為了避免這種情形產生，尋找一種新型的抗菌療法取代傳統的抗生素療法是一重要的課題，而光動力治療法(photodynamic therapy, PDT)被認為是可以有效防止抗藥性細菌的生成，近幾年以來，有越來越多的學者投入此領域研究。

本研究利用同軸靜電紡絲技術生產具拉伸性的奈米纖維膜並結合光動力療法應用於傷口敷料上，主要可分為四大部分：第一部分利 用一種抗沾黏的材料 [2-(methacryloyloxy)ethyl] dimethyl -(3-sulfopropyl) ammonium hydroxide (SBMA)與具有化學交聯反應的單體N-methylol acrylamide (NMA)以自由基聚合法合成poly(SBMA-co-NMA)共聚高分子及其性質鑑定；第二部分利用同軸靜電紡絲技術製備以熱塑性聚氨酯彈性橡膠(thermoplastic polyurethane, TPU)為內層芯材， poly(SBMA-co-NMA)共聚高分子為外層鞘材的同軸芯鞘型纖維膜，並將親水性帶正電光敏劑亞甲基藍(methylene blue, MB)分別混摻至芯層與鞘層，且進行加熱交聯以得到結構穩定的奈米纖維膜，實驗證實以120℃交聯72 hr的纖維膜具有較高的結構穩定性；第三部分則針對傷口敷料進行相關的性質鑑定，同軸纖維膜的水氣滲透率為2110.75 g.m-2day-1符合理想傷口敷料水氣滲透率範圍2000-2500 g.m-2day-1。並藉由拉伸試驗證實同軸芯鞘型纖維膜可以改善poly(SBMA-co-NMA)單層膜的機械性質。另外亦討論光敏劑在核層與殼層的釋放情形，由UV-Vis結果得知，位於殼層的親水性光敏劑較容易釋放，並透過2',7'-dichlorodihydrofluorescein diacetate (DCFH-DA)試劑證實纖維膜具有光動力效果；第四部分為抗菌實驗：以革蘭氏陰性菌和革蘭氏陽性菌的抑菌圈與細菌存活率來檢測纖維膜的抗菌效果。

Antibiotics misuse often results in the formation of drug-resistant bacteria that causes a severe threat to human. In order to prevent this situation, an alternative antibacterial therapy is highly demanded to replace traditional antibiotic therapy. Photodynamic therapy (PDT) is considered to be effective in preventing the formation of drug-resistant bacteria. In recent years, more and more studies have been focused on this field.

In this study, we used the coaxial electrospinning technology to produce stretchable nanofibrous membranes and combined with photodynamic therapy to develop an antibacterial wound dressing. This study was divided into four parts: (I) an anti-fouling material [2- (methacryloyloxy) ethyl] dimethyl-(3-sulfopropyl) ammonium hydroxide (SBMA) and crosslinkable N-methylol acrylamide (NMA) was synthesized by free radical polymerization to form poly(SBMA-co-NMA). (II) a coaxial electrospinning technology was used to prepare core-shell nanofibers with thermoplastic polyurethane (TPU) as the core and poly (SBMA-co-NMA) as the shell, and the photosensitizers, methylene blue (MB), was doped into the core layer or the shell layer. The prepared core-shell nanofibers were thermal cross-linked to obtain a structurally stable nanofiber membrane at 120 curing temperature for 72 hours. (III) We tested the relevant properties for application in wound dressing. The water vapor permeability of the prepared coaxial nanofibrous membrane was 2110.75 g.m-2day-1, which is within the range for applying in ideal would dressing (2000-2500 g.m-2day-1). And the tensile test proves that the coaxial core-shell nanofiber membrane can improve the mechanical properties of poly(SBMA-co-NMA) monolayer membrane. In addition, the release behavior of the photosensitizers doped in the core layer and shell layer was also discussed. The coaxial nanofibers with photosensitizers doped into the shell layer showed faster release rate than doped into the core layer. We also confirmed the coaxial nanofibers with photosensitizers doped had photodynamic therapy effect by using the 2',7'-dichlorodihydrofluorescein diacetate (DCFH-DA) reagent. (IV) the antibacterial effects on gram-negative bacteria and gram-positive bacteria will be tested.

致謝 II
中文摘要 III
Abstract V
目錄 VII
表目錄 XII
圖目錄 XIII
第一章 序論 1
1.1 前言 1
第二章 文獻回顧 3
2.1 靜電紡絲簡介 3
2.1.1 靜電紡絲原理 4
2.1.2 靜電紡絲參數 5
2.1.3 靜電紡絲分類 8
2.2 靜電紡絲應用於傷口敷料 10
2.3 抗沾黏材料 11
2.3.1 發展歷程 13
2.3.2 第三代雙離子性材料 15
2.4 交聯聚合物 18
2.4.1 N-羥甲基丙烯酰胺 (N-methylol acrylamide) 19
2.5 光敏劑 (Photosensitizer, PS) 20
2.5.1 亞甲基藍 (Methylene blue, MB) 25
2.6 光動力治療 26
2.7 研究動機與目的 28
第三章 實驗方法及步驟 30
3.1 實驗架構 30
3.2 poly(SBMA-co-NMA)之聚合 31
3.2.1 實驗材料與藥品 31
3.2.2 實驗裝置與設備 31
3.2.3 實驗步驟 32
3.2.4 實驗檢測儀器 34
3.3 利用靜電紡絲技術製備奈米纖維膜 34
3.3.1 實驗材料及藥品 34
3.3.2 實驗裝置及設備 35
3.3.3 實驗步驟 36
3.3.4 實驗檢測儀器 38
3.4 電紡纖維型態 39
3.5 交聯溫度對材料之影響 39
3.5.1 材料熱性質及熱穩定分析 40
3.5.2 芯鞘型纖維膜對水的穩定性 42
3.6 纖維性質鑑定 43
3.6.1 纖維親疏水鑑定 43
3.6.2 纖維吸水率 (Water uptake ratio) 44
3.6.3 水氣保留率 (Water residual rate) 45
3.6.4 水氣滲透率 (Water vapor permeability rate, WVTR) 46
3.6.5 抗生物沾黏 47
3.7 纖維機械性質 49
3.7.1 實驗步驟 50
3.7.2 實驗檢測儀器 50
3.8 纖維包藥檢測 50
3.8.1 藥物包覆率 (Loading content, L.C.)/藥物封裝效率(Encapsulation efficiency, E.E.) 51
3.8.2 藥物釋放 51
3.9 活性氧分子 (Reactive oxygen species, ROS)檢測 52
3.9.1 實驗材料與藥品 52
3.9.2 實驗裝置 52
3.9.3 實驗步驟 53
3.9.4 實驗檢測儀器 54
3.10 體外細胞毒性測試 55
3.10.1 實驗材料與藥品 55
3.10.2 實驗裝置與設備 56
3.10.3 實驗步驟 56
3.10.4 實驗檢測儀器 59
3.11 抗菌測試 59
3.11.1 實驗材料與菌種 59
3.11.2 實驗裝置 60
3.11.3 抑菌圈實驗 60
3.11.4 細菌存活率檢測 63
第四章 結果與討論 64
4.1 poly(SBMA-co-NMA)共聚高分子之合成與鑑定 64
4.2 同軸芯鞘型纖維之鑑定 65
4.3 電紡纖維型態 67
4.4 交聯溫度對材料之影響 73
4.4.1 材料熱穩定分析 73
4.4.2 纖維膜對水的穩定度檢測 76
4.5 纖維性質鑑定 80
4.5.1 纖維親疏水鑑定 80
4.5.2 纖維吸水率 (Water uptake ratio) 81
4.5.3 水氣保留率 (Water residual ratio) 82
4.5.4 水氣滲透率 (Water vapor permeability rate, WVTR) 83
4.5.5 抗生物沾黏 84
4.6 纖維機械性質 86
4.6.1 應力-應變曲線 86
4.6.2 拉伸後同軸芯鞘型纖維(未包藥)型態 88
4.7 纖維包藥測試 89
4.7.1 藥物包覆率 (Loading content, L.C.)/藥物封裝效率(Encapsulation efficiency, E.E.) 89
4.7.2 藥物釋放 91
4.8 活性氧分子 (Reactive oxygen species, ROS)檢測 93
4.9 生物相容性 95
4.10 抗菌測試 97
4.10.1 抑菌圈測試 97
4.10.2 細菌存活率測試 102
第五章 結論與未來展望 106
參考文獻 108

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