- 學位論文
膠原蛋白/幾丁聚醣/硫酸軟骨素做為人工皮膚基材之研發
Artificial skin development: preliminary studies of a composite matrix
摘要
目前在組織工程的發展方面，仍以人工皮膚的研發最為純熟，本 研究欲建立一個皮膚替代物的立體模型來取代一般的體外細胞毒性試驗，以減少實驗動物的犧牲與龐大經濟成本的支出。本實驗主要分為四個階段:第一階段，萃取來自豬皮的第一型膠原蛋白。第二階段，將膠原蛋白溶液加入幾丁聚醣和硫酸軟骨素，以冷凍乾燥的方式製備出多孔性海綿狀基質。第三階段主要是材料性質分析，以熱穩定分析儀（示差掃描熱卡計）、傅立葉轉換紅外光分光光譜儀以及動態分析儀對基質的基本性質進行分析，並配合掃描式電子顯微鏡的結果作一初步探討。第四階段進行細胞培養，以初級培養的人類纖維母細胞和角質細胞分別於基質中進行培養，將培養兩個星期的基質做H & E染色(Hematoxylin and eosin stain；H & E stain)切片並配合電子顯微鏡觀察細胞於基質中生長的情形。
並列摘要
So far, the most established system in tissue engineering is the development of the artificial skin. In order to reduce the number of animals used in trials, an in vitro model would be helpful. In this study, the idea is to create a skin substitute that could mimic the real skin by first develop a composite matrix for cell and tissue culture. There are four parts in this plan. First, obtain the type I collagen from porcine skin. Second, prepare a 3-D, sponge-liked (porous) collagen based matrix with chitosan and chondroitin sulfate. Third, investigate the physicochemical properties of the prepared material by Differential Scanning Calorimetry (DSC), Fourier-Transform Infrared (FTIR) spectroscopic studies, Dynamic Mechanical Analysis (DMA) and Scanning Electron Microscopic (SEM) studies. Fourth, culture the human dermal fibroblasts and epidermal keratinocytes into the matrix and to observe the cell growth and proliferation by SEM and H & E stain.