Uchida Noriyuki, Sivaraman Srikanth, Amoroso Nicholas J, Wagner William R, Nishiguchi Akihiro, Matsusaki Michiya, Akashi Mitsuru, Nagatomi Jiro
Department of Chemistry and Biotechnology, School of Engineering, the University of Tokyo, 7-3-1 Hongo, Bunkyo-Ku, Tokyo, 113-8656, Japan.
RIKEN Center for Emergent Matter Science, 2-1 Hirosawa, Wako, Saitama, 351-0198, Japan.
J Biomed Mater Res A. 2016 Jan;104(1):94-103. doi: 10.1002/jbm.a.35544. Epub 2015 Aug 6.
Surface modification can play a crucial role in enhancing cell adhesion to synthetic polymer-based scaffolds in tissue engineering applications. Here, we report a novel approach for layer-by-layer (LbL) fabrication of nanometer-size fibronectin and gelatin (FN-G) layers on electrospun fibrous poly(carbonate urethane)urea (PCUU) scaffolds. Alternate immersions into the solutions of fibronectin and gelatin provided thickness-controlled FN-G nano-layers (PCUU(FN-G) ) which maintained the scaffold's 3D structure and width of fibrous bundle of PCUU as evidenced by scanning electron miscroscopy. The PCUU(FN-G) scaffold improved cell adhesion and proliferation of bladder smooth muscles (BSMCs) when compared to uncoated PCUU. The high affinity of PCUU(FN-G) for cells was further demonstrated by migration of adherent BSMCs from culture plates to the scaffold. Moreover, the culture of UROtsa cells, human urothelium-derived cell line, on PCUU(FN-G) resulted in an 11-15 μm thick multilayered cell structure with cell-to-cell contacts although many UROtsa cells died without forming cell connections on PCUU. Together these results indicate that this approach will aid in advancing the technology for engineering bladder tissues in vitro. Because FN-G nano-layers formation is based on nonspecific physical adsorption of fibronectin onto polymer and its subsequent interactions with gelatin, this technique may be applicable to other polymer-based scaffold systems for various tissue engineering/regenerative medicine applications.
在组织工程应用中,表面改性对于增强细胞与合成聚合物基支架的粘附性可能起着至关重要的作用。在此,我们报告了一种在电纺聚(碳酸酯聚氨酯)脲(PCUU)纤维支架上逐层(LbL)制备纳米级纤连蛋白和明胶(FN-G)层的新方法。交替浸入纤连蛋白和明胶溶液中可提供厚度可控的FN-G纳米层(PCUU(FN-G)),扫描电子显微镜显示,该纳米层保持了PCUU支架的三维结构和纤维束宽度。与未涂层的PCUU相比,PCUU(FN-G)支架改善了膀胱平滑肌细胞(BSMCs)的粘附和增殖。粘附的BSMCs从培养板迁移到支架上,进一步证明了PCUU(FN-G)对细胞具有高亲和力。此外,在PCUU(FN-G)上培养人尿路上皮来源的细胞系UROtsa细胞,形成了具有细胞间接触的11 - 15μm厚的多层细胞结构,尽管许多UROtsa细胞在PCUU上未形成细胞连接就死亡了。这些结果共同表明,这种方法将有助于推进体外工程化膀胱组织的技术。由于FN-G纳米层的形成基于纤连蛋白在聚合物上的非特异性物理吸附及其随后与明胶的相互作用,该技术可能适用于其他基于聚合物的支架系统,用于各种组织工程/再生医学应用。