Salerno Simona, Piscioneri Antonella, Laera Stefania, Morelli Sabrina, Favia Pietro, Bader Augustinus, Drioli Enrico, De Bartolo Loredana
Institute on Membrane Technology, National Research Council of Italy, ITM-CNR, c/o University of Calabria, via P. Bucci cubo 17/C, I-87030 Rende (CS), Italy.
Biomaterials. 2009 Sep;30(26):4348-56. doi: 10.1016/j.biomaterials.2009.04.052. Epub 2009 Jun 4.
PEEK-WC-PU membranes were modified with an NH(3) glow discharge process to graft N-containing functional groups at their surface in order to improve the maintenance of human hepatocytes. Native and modified membrane surfaces were characterized with XPS, ToF-SIMS and WCA measurements. We have investigated morphological behaviour and specific functions of primary human hepatocytes on native and modified PEEK-WC-PU membranes in a small-scale gas-permeable bioreactor. N-containing groups grafted at the surface of the membranes improved the initial steps of adhesion and the maintenance of phenotype and differentiated functions of cells. Confocal microscopy of cell morphology evidenced human hepatocytes exhibiting a polygonal shape and organizing a 3D structure. The presence of CK19 positive cells, a marker of biliary duct epithelium, was also found on native and modified membranes. Liver specific functions, investigated in terms of urea production, albumin synthesis and diazepam biotransformation, were maintained at high levels up to 19 days, particularly on surface-modified membranes.
采用NH₃辉光放电工艺对聚醚醚酮-木质素-聚氨酯(PEEK-WC-PU)膜进行改性,以在其表面接枝含氮官能团,从而改善人肝细胞的维持情况。通过X射线光电子能谱(XPS)、飞行时间二次离子质谱(ToF-SIMS)和水接触角(WCA)测量对天然和改性膜表面进行了表征。我们在小型透气生物反应器中研究了原代人肝细胞在天然和改性PEEK-WC-PU膜上的形态行为和特定功能。膜表面接枝的含氮基团改善了细胞黏附的初始步骤以及细胞表型和分化功能的维持。细胞形态的共聚焦显微镜观察表明人肝细胞呈现多边形形状并形成三维结构。在天然和改性膜上还发现了胆管上皮标志物细胞角蛋白19(CK19)阳性细胞。从尿素生成、白蛋白合成和地西泮生物转化方面研究的肝脏特定功能在长达19天的时间内保持在高水平,特别是在表面改性膜上。