具表面構形之聚電解質多層膜於細胞排列操控之應用

Patterned polyelectrolyte multilayer surface for the manipulation of cell alignment

陳建安(Chien-An Chen)

指導教授 ： 孫一明

元智大學/工程學院/化學工程與材料科學學系/碩士(2004年)

摘要

本研究利用軟蝕刻(soft lithography)技術，於基材表面產生微米級構形，藉此操控細胞之貼附與生長。首先，以光蝕刻(photolithography)技術分別於不鏽鋼板與矽晶片上蝕刻出凹、凸構形，再以此材料作為母模並利用poly(dimethylsiloxane) (PDMS)矽膠轉印其反構形；該反轉印後之矽膠可作為微觸印刷(microcontact printing, μCP)之印章，其構形解析度可達10 μm。
結合layer-by-layer (LBL)與μCP技術，可將相反電荷之聚電解質—poly(allylamine hydrochloride) (PAH)與 poly(acrylic acid) (PAA)逐層吸附於蓋玻片表面，形成6—10層多層膜，並在其最外層產生PAH/PAA交替之化學構形；再藉由化學反應之特異性，將能與胺基(-NH2)反應之分子接枝於PAH區域，並露出親水端的聚乙二醇(polyethylene glycol, PEG)，最終則在基材表面生成PAA與PEG交錯之化學構形。以此具表面化學構形之基材，在血清培養環境下，操控C3A細胞選擇性地貼附於PAA之表面，將細胞侷限於50 μm 線寬之區間。

關鍵字

軟蝕刻 ； 聚電解質 ； 聚乙二醇 ； C3A細胞 ； 微觸印刷

並列摘要

In this study, soft lithography technique was applied to create micropatterns on substrate surface for the manipulation of cell adhesion and growth. Micropatterns were first fabricated on stainless steel plates or silicon wafers via photolithographic technique. By using these topo- logically-patterned substrates as masters, complementary poly(dimethyl- siloxane) (PDMS) replicas were prepared. The PDMS replicas were used as pattern transfer elements (stamps) in microcontact printing (mCP), and their spatial resolution could reach 10 mm. Combining layer-by-layer (LBL) and mCP techniques, oppositely charged polyelectrolytes, poly(allylamine hydrochloride) (PAH) and poly(acrylic acid) (PAA), were alternatively deposited onto cover glasses for 6 to 10 layers, and chemically differentiated patterns of PAH/PAA were on the topmost layer. Hydrophilic polyethylene glycol (PEG) moiety was grafted onto the PAH region through PEGylation with an amine-reactive coupling reaction. Finally, the topmost layer became PEG/PAA patterned surface. C3A cells selectively adhered and grew on the PAA region of the patterned surface in serum-containing medium and the spatial resolution could reach 50μm.

並列關鍵字

soft lithography ； polyelectrolyte ； polyethylene glycol (PEG) ； C3A cell ； microcontact printing (mCP) ； layer-by-layer (LBL)

參考文獻

Adams, J. C., “Cell adhesion-spreading frontiers, intricate insight,” Trends Cell Biol., 7, 107 (1997)

Benjamin, H. S., L. Li and P. Yoon, “Thymic peptide protects vascular endothelial cells from hydrogen peroxide-induced oxidant injury,” Life Sci., 52, 1787 (1993)

Bhatia, S. N., M. L. Yarmush, and M. Toner, “Controlling cell interactions by micropatterning in co-cultures: Hepatocytes and 3T3 fibrolasts,” J. Biomed. Mater. Res., 34, 189 (1997)

Blawas, A. S., and W. M. Reichert, “Protein patterning,” Biomaterials, 19, 595 (1998)

Caruso, F., H. Lichtenfeld, E. Donath, and H. Möhwald, “Investigation of electrostatic interactions in polyelectrolyte multilayer films: binding of anionic fluorescent probes to layers assembled onto colloids,” Macromolecules, 32, 2317 (1999)