A novel method to immobilize collagen on polypropylene film as substrate for hepatocyte culture
A novel method to immobilize collagen on polypropylene film as substrate for hepatocyte culture
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DOI：
10.1016/j.msec.2005.08.039
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发表时间：
2006-05
期刊：
影响因子：
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通讯作者：
Yong Zhang;Wenjie Wang;Q. Feng;F. Cui;Ying-xin Xu
中科院分区：
文献类型：
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作者：
Yong Zhang;Wenjie Wang;Q. Feng;F. Cui;Ying-xin Xu
为提高聚丙稀膜与肝细胞的生物相容性，首先通过过氧二硫酸铵(APS)热分解生成过氧化氢基团，然后以丙烯酸为单体，亚铁离子为氧化还原引发剂，进行自由基接枝聚合。然后在水溶性碳二亚胺存在下，通过残基胺和羧基之间的酰胺键共价固定胶原蛋白。用衰减全反射傅里叶变换红外光谱(ATR-FTIR)、X-光电子能谱(XPS)和激光共聚焦扫描显微镜(CLSM)等手段对薄膜进行了表征，实验结果表明，APS水热分解后存在过氧化氢基团，并转化为引发乙烯基单体进行接枝聚合的聚合物自由基。最大固定化胶原量为10.5μg/cm2。扫描电子显微镜(SEM)和生化参数测定显示，在胶原固定化薄膜上培养的肝细胞在20天以上表现出稳定的表型和正常的肝功能。
To improve the biocompatibility of polypropylene membrane with hepatocytes, film was firstly allowed to generate peroxide groups by ammonium peroxydisulfate (APS) thermal decomposition and was then carboxylated by free radical grafting polymerization using acrylic acid as the monomer and ferrous(II) ions as redox initiator. Collagen was then covalently immobilized through amide bonds between the residue amine groups and carboxylic ones in the presence of water-soluble carbodiimide. The film was characterized by Fourier transformed infrared spectroscopy in attenuated total reflection mode (ATR–FTIR), X-photoelectron spectroscopy (XPS) and confocal laser scanning microscopy (CLSM), etc. The experimental results show that there exist hydroperoxide groups after APS aqueous thermal decomposition, which are subsequently converted into the polymeric free radicals initiating the vinyl monomers to subject grafting polymerization. The maximum collagen immobilization content is about 10.5 μg/cm2by ninhydrin assay. Hepatocytes cultured on collagen immobilized film show stable phenotype and normal liver-specific functions more than 20 days as observed by scanning electronic microscopy (SEM) and biochemical parameters determination.