臺灣博碩士論文加值系統

本研究主要是欲以電紡織法所製得的奈米纖維膜應用在軟骨組織工程中，發展出一套能用於軟骨組織修復的方法。在實驗設計上我們是希望能以有利於細胞分裂生長的成分混合在製備奈米纖維膜的高分子溶液中，而直接得到一個具有高表面積均勻的混有各種能促進細胞生長的成分。
在不同高分子薄膜上培養軟骨細胞的實驗中，我們初步發現材料成分中含有PEO(polyethylene oxide)時，會造成細胞無法很好在支架上的貼附生長。而在奈米纖維薄膜上培養軟骨細胞時，我們也發現此一奈米纖維結構也可能會使得細胞貼附的不穩固而造成細胞無法在膜上正常的生長。
當奈米纖維膜有膠原蛋白吸附修飾後，軟骨細胞在膜上的貼附生長就有比較良好的效果，貼附效率能提升5％ ~ 27％。
綜合以上研究結果，我們發現似乎直接將膠原蛋白混入電紡織成膜的製程中比較沒有明顯的效果，仍需在製成膜後進一步地在膜表面進行修飾才能有個良好的效果。

The aim of this research is to fabricate nano-fibrous membranes from electrospinning for cartilage tissue engineering. Nanofibers were produced by electrospinning of a polymer solution of chitosan, collagen and polyethylene oxide (PEO) and crosslinked by glutaraldehyde vapor. Chondrocytes were isolated from rabbit articular cartilage and seeded on the nanofibrous scaffolds.

By seeding chondrocytes and meniscus cells on film surfaces containing different combinations of collagen, chitosan, and PEO, we found that cells did not attach and proliferate well to films containing PEO. When cells were cultured on nano-fibrous membranes, the nano-fibrous structure may lead to weak cell attachment and poor proliferation.

After type I collagen modification, the nano-fibrous membrane becomes better for cell attachment. The cells attachment efficiency can be improved by 5 ~ 27％ by growing the cells on the collagen modified nano-fibrous membrane.

第一章 緒論 1
1.1 研究動機 1
1.2 研究目的 3
第二章 文獻回顧 5
2.1目前關節的治療修復方式 5
2.1.1 顯微骨折手術 5
2.1.2 異體移植 7
2.1.3 自體移植 9
2.1.4 自體軟骨細胞移植 11
2.1.5 組織工程技術 12
2.2 電紡織技術 18
第三章 實驗設備、藥品及方法 20
3.1 實驗設備 20
3.2 實驗藥品 22
3.3 實驗方法 24
3.3.1 初代軟骨細胞之取得 24
3.3.2 繼代培養 25
3.3.3 軟骨細胞數測定 26
3.3.4 24孔細胞培養盤預處理 27
3.3.5 軟骨細胞MTT測定 28
3.3.6 細胞外間質-葡萄糖胺聚醣（GAG）分析 30
3.3.7掃描式電子顯微鏡（SEM）樣品前處理 32
3.3.8奈米纖維膜的交聯 33
3.3.9奈米纖維膜及高分子塗佈薄膜表面的修飾處理 35
第四章 初步結果與討論 36
4.1 初代軟骨細胞之取得 36
4.2 利用光學顯微鏡觀察軟骨細胞活性及其型態 40
4.3 細胞培養盤（TCPS）平面培養細胞活性測試 42
4.4軟骨細胞在奈米纖維膜上培養的活性測試 44
4.5 軟骨細胞在高分子薄膜（FILM）上生長情形 47
4.6軟骨細胞在奈米纖維膜及高分子薄膜上的生長與分泌測試 51
4.7軟骨細胞在第一型膠原蛋白（TYPE I COLLAGEN）修飾後的奈米纖維膜及高分子薄膜上之生長測試 51
第五章 結論 68
第六章 參考文獻 70

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