臺灣博碩士論文加值系統

本研究探討溫度敏感性甲基纖維素水溶液之相轉移情形和成膠動力學，並測試間葉幹細胞在甲基纖維素水膠中之生物相容性。添加0.13 M NaCl至甲基纖維素水溶液可產生鹽析作用，而降低成膠溫度及提升成膠速率。高濃度甲基纖維素溶液，因鏈間糾纏造成相轉移溫度下降及成膠速率提高。甲基纖維素溶液至9 wt%以上，成膠溫度趨近於42℃，而藉由鹽類添加造成成膠溫度降低而趨近至40℃。在細胞培養部分，當間葉幹細胞培養於3D甲基纖維素水膠中，可觀察到細胞聚集現象，並且能夠存活至28天，證實甲基纖維素水膠具生物相容性，為可應用於組織工程之注射型水膠。

The phase transition behaviors and the gelation kinetics of the thermosensitive methylcellulose (MC) solution were studied and the MC gels were tested for the biocompatibility with mesenchymal stem cells (MSCs). The presence of NaCl (0.13 M) decreased the gelation temperature (or cloud point) and increased the rate of the gelation process due to the salting out effect. The gelation temperature decreased and the rate of gelation increased with the increase of MC concentration in the solution due to the chain entanglement effect at higher polymer concentrations. The gelation temperature of the salt-free MC solution approximated to 42 oC and that of the MC solution with 0.13 M NaCl approximated to 40 oC when the polymer concentration was higher than 9 wt%. When MSCs were cultured within the 3D MC gels, it was found that MSCs could aggregate and maintain their viability up to 28 days. The biocompatibility of MC gels with MSCs suggested that MC gels could be considered as an injectable hydrogel for tissue engineering application.

致謝 I
摘要 II
Abstract III
目錄 IV
表目錄 VIII
圖目錄 IX
第一章 序論 1
1-1 水膠( Hydrogel ) 1
1-2 組織工程 1
1-3 支架( Scaffold ) 3
1-4 研究動機與範疇 4
第二章 文獻回顧 5
2-1 溫度敏感性水膠( Thermo-sensitive hydrogels ) 5
2-2 溫度敏感性水膠—甲基纖維素( Methylcellulose ) 8
2-3 溫度敏感性甲基纖維素成膠機制 10
2-4 鹽類對溫度敏感性甲基纖維素的影響 12
2-5 水膠在組織工程之應用 13
2-6 間葉幹細胞在膠裡的生長情形 14
2-7 間葉幹細胞與組織工程之應用 14
第三章 材料與實驗方法 16
3-1實驗藥品 16
3-2 實驗設備 17
3-3溶液配製 18
3-3-1 配製16 wt%的甲基纖維素溶液( Methylcellulose solution ) 18
3-3-2 DMEM F-12培養基 ( DMEM F-12 medium ) 19
3-3-3 磷酸緩衝溶液( Phosphate buffer solution, PBS ) 19
3-3-4 Blue CMAC試劑(7-amino-4-chloromethylcoumarin, CMAC) 19
3-3-5 MTT試劑( 3-(4,5-dimethylthiazol-2-yl )-2,5-
diphenyltetrazolium Bromide ) 20
3-4 基材分析 20
3-4-1 甲基纖維素濃度初步測試 23
3-4-2 光穿透實驗( UV-Vis spectrophotometer )[Zheng et al.,
2004]熱性質分析 24
3-4-3 溫度敏感性甲基纖維素溶液雲點分佈 25
3-4-4 光穿透實驗動力學分析 25
3-4-5 溫度敏感性甲基纖維素溶液動力學分析 26
3-4-6 倒立式顯微鏡 26
3-5 細胞培養實驗 27
3-5-1 間葉幹細胞( MSCs )培養 27
3-5-2 細胞與基材測試 28
3-5-3 細胞型態觀察 29
3-5-4 CMAC (7-amino-4-chloromethylcoumarin, CMAC )染劑 29
3-5-5 MTT測試細胞活性 30
第四章 結果與討論 32
4-1 溫度敏感性甲基纖維素溶液熱性質分析 32
4-2 溫度敏感性甲基纖維素溶液之雲點分布 32
4-3 溫度敏感性甲基纖維素溶液動力學分析 33
4-4 溫度敏感性甲基纖維素溶液動力學分析 34
4-5 溫度敏感性甲基纖維素溶液黏度分析 35
4-6 溫度敏感性甲基纖維素溶液構形分析 35
4-7 溫度敏感性甲基纖維素溶液成膠性 36
4-8 間葉幹細胞與溫度敏感性甲基纖維素溶液之培養前測試 36
4-8-1 溫度敏感性甲基纖維素內含0.13 M NaCl鹽類之膠體 36
4-8-2 溫度敏感性甲基纖維素溶液內含1倍PBS緩衝溶液鹽類之膠體 37
4-8-3 間葉幹細胞培養在溫度敏感性甲基纖維素內含1倍培養液及
0.13 M NaCl鹽類 38
4-8-4 間葉幹細胞培養在溫度敏感性甲基纖維素水膠的活性測試 39
第五章 結論 66
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