開發促進傷口癒合之複合敷料

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姓名

林于廷(Yu-Ting Lin) 查詢紙本館藏

畢業系所

化學工程與材料工程學系

論文名稱

開發促進傷口癒合之複合敷料
(The Development of Composite Dressings to Promote Wound Healing)

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摘要(中)

慢性傷口癒合緩慢，可能會造成很多風險與生活上的不便，因此本研究欲開發多功能傷口敷料以促進組織再生。我們以雙噴射電紡絲系統將褐藻酸鈉與聚己內酯（poly (ε-caprolactone)，PCL）混紡成多功能複合納米纖維。其中親水性的褐藻酸鈉纖維具高吸收性，可以在傷口部位提供潮濕環境，PCL則可增加機械強度並促進細胞貼附。將奈米銀導入PCL纖維中使其可以持續抑制微生物的生長。由於血小板衍生生長因子（platelet-derived growth factor B，PDGFB）是嗜中性粒細胞的化學引誘物並且可以誘導成纖維細胞的增殖和分化，進而促進傷口修復，因此我們將PDGFB質粒DNA與聚乙烯亞胺（PEI）所複合成的正電性奈米顆粒通過靜電相互作用吸附於複合纖維中的陰離子褐藻酸鈉纖維上。當傷口細胞貼附到複合纖維時將被原位轉染，而這些被轉染的細胞將可連續表達PDGFB。此外，褐藻酸鈉纖維中的鈣離子通過離子交換釋放到傷口部位以加速止血。這種全面的敷料為慢性傷口的治療提供了理想的解決方案。

摘要(英)

Chronic wounds may retard the healing process to cause many risks. Therefore, it is essential to develop a multifunctional wound dressing to promote tissue regeneration. To fabricate a versatile composite nanofibrous matrix, sodium alginate and poly (ε-caprolactone) (PCL) were coelectrospun as composite nanofibers using a dual jet system. Hydrophilic alginate fibers may provide a moist environment in wound sites. In addition, PCL were applied to increase mechanical strength and cell adhesion. Silver nanoparticles were embedded in PCL fibers for long-term release to inhibit the growth of microorganism. Plasmid DNA encoding platelet-derived growth factor B(PDGFB) was delivered from composite fibers because this growth factor is a chemoattractant for neutrophils and can induce the proliferation and differentiation of fibroblasts. These PDGFB plasmids were complexed with polyethylenimine (PEI) to form cationic nanoparticles which may thus be adsorbed onto anionic alginate fibers through electrostatic interaction. As wound cells adhered to composite fibers, they can be in situ transfected to continuously express PDGFB. Moreover, calcium ions in alginate fibers were released to wound sites through ion exchange to accelerate hemostasis. This comprehensive dressing provides an ideal solution to heal chronic wounds.

關鍵字(中)

★ 電紡絲
★ 傷口敷料
★ 奈米銀
★ 血小板衍生生長因子B

關鍵字(英)

論文目次

摘要 v
Abstract vi
致謝 vii
目錄 viii
圖目錄 xi
表目錄 xiv
第一章緒論 1
1-1研究背景 1
1-2研究目的 3
第二章文獻回顧 4
2-1電紡絲 4
2-1-1電紡絲原理 4
2-1-2 製備複合電紡絲 5
2-1-3 複合電紡絲的應用 7
2-2 褐藻酸鈉 10
2-2-1 褐藻酸鈉之簡介 10
2-2-2 褐藻酸鈉之性質 10
2-2-3 褐藻酸鈉的應用 14
2-3 聚己內酯 16
2-3-1聚己內酯之簡介 16
2-3-2聚己內酯之性質 17
2-3-3 聚己內酯的應用 17
2-4 奈米銀 19
2-4-1奈米銀之簡介 19
2-4-2奈米銀之抗菌功能 20
2-4-3奈米銀於傷口敷料的應用 23
2-5調節傷口癒合的生長因子 26
2-6基因治療 28
2-6-1載體 29
2-6-2基因治療的應用 29
第三章實驗藥品與方法 31
3-1實驗原料及藥品 31
3-2實驗儀器 35
3-3實驗方法 37
3-3-1電紡絲溶液製備 37
3-3-2奈米銀製備 38
3-3-3電紡絲纖維製備 39
3-3-4電紡絲纖維收集量之量測 40
3-3-5溶液配製 40
3-3-6 NIH 3T3細胞培養 42
3-3-7 SEM之樣本製備 44
3-3-8TEM樣本製備 46
3-3-9XRD樣本製備 46
3-3-10FTIR/ATR樣本製備 46
3-3-11TGA、DSC樣本製備 46
3-3-12細胞存活率實驗(MTT assay) 47
3-3-13 質體DNA(PDGFB) 純化 48
3-3-14原位轉染實驗 49
3-3-15測量PDGFB釋放實驗 51
3-3-16抗菌實驗 53
3-3-17奈米銀檢測[61] 55
3-3-18凝血實驗[62] 56
3-3-19 傷口癒合動物實驗 57
第四章結果與討論 59
4-1褐藻酸鈉/聚己內酯/奈米銀之複合電紡絲性質 59
4-1-1電紡絲之收集量與直徑分佈 59
4-1-2奈米銀於電紡絲中之型態與分析 65
4-1-3複合電紡絲之收集 73
4-2複合電紡絲之抗菌活性 74
4-2-1液態培養之抗菌活性 74
4-2-2固態培養之抗菌活性 77
4-3複合電紡絲對細胞之影響 79
4-3-1細胞於複合纖維上培養之SEM圖 79
4-3-2細胞於複合纖維上培養之存活率 80
4-4複合電紡絲之奈米銀釋放 82
4-5於複合電紡絲上進行原位轉染PDGFB 85
4-5-1原位轉染之螢光影像 85
4-5-2原位轉染後分析細胞增殖率與PDGFB釋放 87
4-6複合電紡絲之凝血測試 90
4-7 傷口癒合動物實驗 92
第五章結論 96
參考資料 99

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指導教授

胡威文(Wei-Wen Hu)

審核日期

2019-8-20

推文