Preparation of Self-Organized Micro-Patterned Polymer Films Having Cell Adhesive Ligands
Preparation of Self-Organized Micro-Patterned Polymer Films Having Cell Adhesive Ligands
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DOI：
10.1295/polymj.34.166
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发表时间：
2002-03
期刊：
影响因子：
2.8
通讯作者：
J. Nishida;K. Nishikawa;S. Nishimura;S. Wada;T. Karino;Takehiro Nishikawa;K. Ijiro;M. Shimomura
中科院分区：
文献类型：
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作者：
J. Nishida;K. Nishikawa;S. Nishimura;S. Wada;T. Karino;Takehiro Nishikawa;K. Ijiro;M. Shimomura
本文介绍了利用简单铸造法形成的自组织蜂窝结构实现细胞黏附配体微图案化的三种新方法。第一种方法是通过浇注包含作为细胞粘附性配体之一的乳糖残基的两亲性聚合物来直接制备图案化薄膜。将两亲性聚合物的苯溶液在高湿度下浇铸在玻璃基板上。原子力显微镜(AFM)观察表明，薄膜形成了直径为微米级的蜂窝状微孔结构。将薄膜浸泡在荧光标记的凝集素水溶液中，以研究乳糖在图案化薄膜上的分布。凝集素结合膜的荧光图像与蜂窝图案的一致性表明，乳糖残基不存在于孔洞，而是存在于蜂窝图案膜的边缘。第二种方法是通过化学反应将明胶固定在蜂窝图案的薄膜上，明胶也是细胞黏附配体之一。由含有活性琥珀酰亚胺酯基的两亲性聚合物的氯仿溶液制备了蜂窝状薄膜，然后将薄膜浸入荧光标记的明胶水溶液中，在薄膜表面引入明胶。荧光显微镜证实了明胶被固定在蜂窝状薄膜上。第三种方法是通过特定的亲和素-生物素相互作用将明胶引入蜂窝膜的另一种方法。由含有生物素残基和十二基的两亲性聚合物制备了蜂窝状薄膜，然后将其依次浸泡在亲和素溶液和生物素化荧光标记的明胶溶液中。通过对膜的荧光显微镜观察，证实明胶通过亲和素-生物素相互作用被固定在蜂窝状结构的边缘。用第二种方法制备的明胶固定化图案膜进行细胞培养。明胶固定化蜂窝膜的生物活性通过细胞在膜上的粘附性得到证实。
This article describes novel three methods for micro-patterning of cell adhesive ligands by using the self-organized honeycomb-patterned structure formed by the simple cast method. A first method is a direct preparation of a patterned film by casting an amphiphilic polymer containing lactose residue which is one of cell adhesive ligands. A benzene solution of the amphiphilic polymer was cast at high humidity on a glass substrate. Atomic force microscopy (AFM) observation of the film showed that a honeycomb pattern with microporousness with as large as micrometer size in diameter was formed. The film was immersed into an aqueous fluorescence-labeled lectin solution to investigate the distribution of lactoses on the patterned film. Consistence of a fluorescence image of the lectin bound film with the honeycomb pattern showed that the lactose residues were existed not at the holes but at the rims of the honeycomb-patterned film. A second method is to immobilize gelatin, which is one also one of cell adhesive ligands, on the honeycomb-patterned film by chemical reaction. A honeycomb-patterned film was prepared from chloroform solution of an amphiphilic polymer containing reactive succinimide ester groups, and then the film was immersed into an aqueous fluorescence-labeled gelatin solution to introduce gelatin on the film surface. Immobilization of gelatin onto honeycomb-patterned film was confirmed by the fluorescence microscope. A third method is another way to introduce gelatin onto the honeycomb film by the specific avidin-biotin interaction. A honeycomb-patterned film was prepared from amphiphilic polymer containing biotin residues and dodecyl groups, and then the film was immersed into a avidin solution and a biotinylated fluorescence labeled gelatin solution successively. By the fluorescence microscopic observation of the film, gelatin was confirmed to be immobilized at the rims of the honeycomb pattern via the avidin-biotin interaction. Cell culture was performed on the gelatin immobilized patterned film prepared by second method. Bioactivity of gelatin immobilized honeycomb-patterned film was confirmed by adhesion of cell onto the film.