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論文中文名稱:幾丁聚醣薄膜中包覆褐藻酸顆粒以改善褐藻酸薄膜之細胞與藥物傳送效率 [以論文名稱查詢館藏系統]
論文英文名稱:Chitosan film coating alginate beads for improving the cell in alginate beads and promoting the efficiency for drug release [以論文名稱查詢館藏系統]
院校名稱:臺北科技大學
學院名稱:工程學院
系所名稱:化學工程研究所
畢業學年度:103
畢業學期:第二學期
中文姓名:羅婉榕
英文姓名:Lo,Wan-Jung
研究生學號:102738041
學位類別:碩士
口試日期:2015/07/23
指導教授中文名:林忻怡
指導教授英文名:LIN HSIN-YI
口試委員中文名:蔡偉博;王孟菊
中文關鍵詞:幾丁聚醣纖維母細胞褐藻酸鈉顆粒
英文關鍵詞:chitosanfibroblastalginate beads
論文中文摘要:褐藻酸鈉是無毒、水溶性天然高分子因此被廣泛地運用在組織工程中,幾丁聚醣也是常用的天然基材,利用這兩種天然高分子製成複合式材料,實驗利用褐藻酸鈉顆粒包覆藥物改善褐藻酸鈉幾丁聚醣的藥物釋放速率,對照組是包覆細胞的褐藻酸鈉顆粒混入純的褐藻酸鈉薄膜,實驗組是包覆細胞的褐藻酸鈉顆粒混入含有幾丁聚醣顆粒藻酸鈉薄膜。來觀察在不同環境下細胞的生長、細胞增生與活性的差異。
在細胞存活螢光染色中觀察實驗組與對照組的細胞在第一、七天生長沒有太大的差異,但到了第十四天實驗組的細胞存活明顯高於對照組,結果顯示加入幾丁聚醣的複合式材料可改善細胞生長,DNA定量第一天和第四天沒有明顯的生長,隨者時間的增加在第七和第十四天細胞增生實驗組高於對照組,將藥物包在褐藻酸鈉顆粒中和加入幾丁聚醣顆粒增加了藥物釋放的阻力,而實驗組中的幾丁聚醣會與褐藻酸鈉作交聯,可減緩藻酸鈉薄膜的降解速率因此減緩藥物釋放的速率,讓藥物對抑制細胞發炎反應的時間變長。
論文英文摘要:Alginate hydrogels are used extensively in cell encapsulation, non-toxic, and tissue engineering applications. Chitosan is also commonly used natural substrates using both a natural polymer made of composite material. Drugs used in this experiment were embedded in alginate beads to improve the chitosan film rate of drug release . Control group cell was embedded in alginate beads which is coating in pure alginate film. The experimental group is alginate coated beads into cells containing chitosan particles of alginate film to observe the growth of cells in different environments, differences in cell proliferation and activity.
Cells were observed in the experimental group and the control group in Live/Dead staining of cell survival in the day 1,7 is not much difference in the growth, but in the day14 of the experimental group was significantly higher cell survival. The results shows that adding chitosan composite material can be improved cell growth, The cell proliferation in day 1,4 no significant growth, with the increase by the time the 7 and day14 of the experimental group than in the control of cell proliferation group. The drug embedded in alginate beads and added chitosan particles to increase the release of the drug resistance. In the experimental group and chitosan will crosslink alginate, alginate can slow the rate of degradation of the film thus slowing the rate of drug release, allowing the drug to inhibit the cell inflammatory response time becomes longer.
論文目次:目錄
摘要 I
ABSTRACT II
誌謝 IV
目錄 V
表目錄 VIII
圖目錄 X
第一章 緒論 1
1.1 前言 1
1.2研究目的 2
2.1 傷口癒合 3
2.2水凝膠 6
2.3 褐藻酸鈉 8
2.4 幾丁聚醣 10
第三章 實驗材料與方法 13
3.1實驗材料 13
3.1.1細胞來源 13
3.1.2細胞培養用藥品 13
3.1.3實驗藥品 14
3.1.4儀器設備 17
3.1.5藥品及溶液配製 20
3.2實驗方法 23
3.2.1實驗設計 23
3.2.2實驗流程 23
3.2.3 噴霧器架設 24
3.2.3.1 噴霧示意圖 25
3.2.4 製作褐酸鈉顆粒 26
3.2.4.1 褐酸鈉顆粒 26
3.2.4.2 褐藻酸鈉平均顆粒之粒徑 26
3.2.4.3 含細胞褐藻酸鈉顆粒存活 26
3.2.4.4 不同天數褐藻酸鈉顆粒中含細胞存活 27
3.2.5製作褐藻酸鈉薄膜 27
3.2.5.1溶液配製 27
3.2.5.2薄膜製作 27
3.3物理性質測試 29
3.3.1拉伸測試 29
3.3.1.1海藻酸鈉拉伸樣品製備 29
3.3.1.2幾丁聚醣拉伸樣品製備 30
3.3.1.3拉伸示意圖 31
3.3.1.樣品拉伸測試 33
.3.4含鈣鎂PBS降解 34
3.5細胞在支架上分佈 35
3.5.1細胞存活螢光染色 35
3.6共軛焦螢光顯微鏡 35
3.7細胞增生 36
3.8細胞活性測定 37
3.9 膠原蛋白測試 37
3.10 雙氯芬酸鈉藥物釋放 38
3.12統計分析 42
第四章 實驗結果與討論 43
4.1物理性質測試 43
4.1.1顆粒大小的比較 43
4.1.2顆粒大小的比較 44
4.1.3.1褐藻酸鈉顆粒細胞存活染色 45
4.1.3.2不同天數褐藻酸鈉顆粒細胞存活染色 46
4.1.2 含鈣鎂PBS降解 47
4.2拉伸測試 49
4.3細胞存活螢光染色 51
圖4.10 纖維母細胞生長在alginate-alginate與alginate-chitosan水凝膠中,進行 Live/Dead螢光染色,放大倍率為100X;可以觀察到大多數細胞都存活下來,在第十四天,alginate-chitosan組有些許細胞有攤開的趨勢 (如紅色箭頭所示),而alginate-chitosan組的細胞數比alginate-alginate細胞數多,推測細胞在含有幾丁聚醣的薄膜中生長情況有比較好 53
4.4細胞增生 55
4.5細胞活性測定 57
.4.6膠原蛋白測定 58
.4.7 雙氯芬酸鈉藥物釋放 59
4.8 基因表現 60
第五章 結論 63
參考文獻 64
附錄一 DNA定量標準曲線 69
附錄二 COLLAGEN 標準曲線 70
附錄三 DICLOFENAC 標準曲線 71
附錄四 PRIMER 72
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