Department of Chemical and Biochemical Engineering, The University of Western Ontario, London, Ontario, Canada N6A 5B9.
Acta Biomater. 2011 Mar;7(3):1114-25. doi: 10.1016/j.actbio.2010.11.010. Epub 2010 Nov 10.
Tissue engineering scaffolds provide the three-dimensional (3-D) geometry and mechanical framework required for regulating cell behavior and facilitating tissue maturation. Unfortunately, most synthetic scaffolds lack the biological recognition motifs required for seeded cell interaction. In order to impart this recognition, synthetic scaffolds should possess appropriate biological functionality. Here, for the first time, we present a comprehensive study of fibronectin (FN) conjugation onto highly porous 3-D poly(carbonate) urethane scaffolds through grafted poly(acrylic acid) spacers on the urethane backbone. Scanning electron microscopy was used to ensure that the porous structures of the scaffolds were preserved throughout the multiple conjugation steps, and Fourier transform infrared spectroscopy was used to monitor the reaction progress. Toluidine blue staining revealed that increasing acrylic acid concentration and grafting time increased the number of poly(acrylic acid) groups incorporated. High resolution X-ray photoelectron spectroscopy studies of the scaffolds demonstrated an increase in nitrogen and sulfur due to FN conjugation. Immunofluorescence microscopy studies showed an even distribution of conjugated FN on the 3-D scaffolds. Cell culture studies using human coronary artery smooth muscle cells demonstrated that FN-conjugated scaffolds had improved cell attachment and infiltration depth compared with scaffolds without FN conjugation and with those scaffolds on which FN was merely adsorbed.
组织工程支架提供了调节细胞行为和促进组织成熟所需的三维（3-D）几何形状和机械框架。然而，大多数合成支架缺乏用于接种细胞相互作用的生物识别基序。为了赋予这种识别，合成支架应具有适当的生物学功能。在这里，我们首次全面研究了通过在聚氨酯主链上接枝聚（丙烯酸）间隔物将纤连蛋白（FN）接枝到高度多孔的 3-D 聚（碳酸酯）聚氨酯支架上。扫描电子显微镜用于确保支架的多孔结构在多次接枝步骤中得以保留，傅里叶变换红外光谱用于监测反应进展。甲苯胺蓝染色表明，随着丙烯酸浓度和接枝时间的增加，接枝的聚（丙烯酸）基团数量增加。支架的高分辨率 X 射线光电子能谱研究表明，由于 FN 接枝，氮和硫的含量增加。免疫荧光显微镜研究表明，FN 接枝到 3-D 支架上均匀分布。使用人冠状动脉平滑肌细胞的细胞培养研究表明，与未接枝 FN 的支架以及仅吸附 FN 的支架相比，FN 接枝的支架具有更好的细胞附着和渗透深度。
