臺灣博碩士論文加值系統

具微米構形之聚電解質多層膜能控制表面地形尺寸至分子級，非常適合體外培養細胞及生醫材料的研究上。
聚丙烯酸(polyacrylic acid，PAA)與聚丙烯醯胺(poly acrylamide，PAAm)在低酸鹼值時，可由氫鍵形成聚電解質多層膜，而且可被鹼性水洗去。配合上疊氮化物（azide）可在紫外光照射下與聚合物交聯的特性，來製備具微米構形之聚電解質多層膜。
首先將4-疊氮苯胺鹽酸鹽(4-azidoaniline hydrochloride)與聚丙烯酸接枝形成PAAz。接著，在載玻片上製作PAA/PAAm多層膜至4個雙層，再吸附PAAz/PAAm多層膜1個雙層後，繼續堆疊PAA/PAAm。將不同雙層數的此多層膜在等尺寸線寬、間距，週期分別為20 μm、50 μm之平行直線光罩下照射紫外光後，用pH 10.0的二次水洗去未交聯的區域，就形成具微米構形之聚電解質多層膜。
進行細胞實驗前，先在此表面上吸附一層聚離胺酸(poly-L-lysine，PLL)，我們發現MG63、L929細胞會選擇在避光的區域貼附、排列，PC12細胞則是照光、避光區域皆會排列，但其突觸會沿著平行線的方向延伸。本研究結果觀察到細胞在週期50 μm的微米構形多層膜表面上排列情況較週期20 μm佳。
本研究成功地利用光刻法製備可調控細胞貼附的具微米構形之聚電解質多層膜。

The topography of micropatterned polyelectrolyte multilayers can be controlled in molecule scale and is extremely versatile and useful for in vitro studies of cell behavior or biomedical devices.
Polyacrylamide (PAAm) and poly(acrylic acid) (PAA) can be assembled to form polyelectrolyte multilayers via hydrogen-bonding interactions at low pH. The PAA/PAAm films can be rinsed away with alkaline water. Azides, a photo-reactive molecule, can crosslink neighbor hydrocarbon chains under UV irradiation.
First, 4-azidoaniline hydrochloride was grafted on PAA to form PAAz. Then, PAA/PAAm films were fabricated on glass slide for 4 bilayers, followed by deposition of one PAAz/PAAm bilayer before further build-up of PAA/PAAm films. Micropatterned polyelectrolyte multilayers with different numbers of bilayers were fabricated by UV exposure of the multilayers films through photomasks with 20 μm or 50 μm periods of parallel lanes.
Poly-L-lysine was adsorbed on micropatterned films before in vitro cell experiments. We found MG63 and L929 cells adhered to ditch region and regularly arranged along the direction of ditch. On the other hand, PC12 cells extended neurites parallel to the striped pattern, but adhered on both ditch and ridge regions. Cell alignment on the micropatterned films with 50 μm periods was better than 20μm periods.
These works successfully developed a technique utilizing polyelectrolyte multilayers and photolithography technology to prepare micropatterned substrata with controlled surface chemistry for modulation of cell adhesion.

誌謝 I
摘要 II
Abstract III
目錄 V
圖目錄 VIII
表目錄 XI
第一章 緒論 1
1.1 研究背景 1
1.2 聚電解質多層膜簡介 3
1.3 具表面微圖案聚電解質多層膜之製備 5
1.4 芳香基疊氮化物（Aryl azide）簡介 8
1.5 細胞株簡介 9
1.6 研究動機 10
1.7 研究架構 10
第二章 實驗藥品、儀器與方法 21
2.1 實驗藥品 21
2.1-1 製備聚電解質多層膜 21
2.1-2 聚丙烯酸接枝疊氮化物 21
2.1-3 聚電解質多層膜染色 21
2.1-4 細胞培養 22
2.2 實驗儀器及耗材 23
2.2-1 實驗儀器 23
2.2-2 實驗耗材 24
2.3 溶液配置 24
2.4 實驗方法 27
2.4-1 PAAz合成與分析 27
2.4-2 聚電解質多層膜製備與分析 28
2.4-3 細胞培養 32
2.4-4 細胞解凍與冷凍 33
2.4-5 多層膜表面上之細胞計數 34
第三章 實驗結果與討論 35
3.1 PAAz的製備與接枝比例分析 35
3.2 探討光可交聯之多層膜的製備起因 37
3.3 H[(A/m)4(z/m)]n多層膜之接觸角分析 38
3.4 H[(A/m)4(z/m)]n多層膜之吸附質量分析 39
3.5 H[(A/m)4(z/m)]n多層膜表面微圖案以甲基藍染色結果
分析 41
3.6 於具表面微圖案之H[(A/m)4(z/m)]n多層膜上培養MG63
細胞並觀察細胞排列情形 42
3.7 於具表面微圖案之H[(A/m)4(z/m)]n多層膜上培養L929
細胞並觀察細胞排列情形 43
3.8 於具表面微圖案之H[(A/m)4(z/m)]n多層膜上培養PC12
細胞並觀察細胞排列情形 43
第四章 結論與未來展望 69
參考文獻 73
附錄 75

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