Interfacing biomembrane mimetic polymer surfaces with living cells – Surface modification for reliable bioartificial liver
Interfacing biomembrane mimetic polymer surfaces with living cells – Surface modification for reliable bioartificial liver
复制标题
DOI：
10.1016/j.apsusc.2008.06.152
复制
发表时间：
2008-11
影响因子：
6.7
通讯作者：
Y. Iwasaki;Utae Takami;S. Sawada;K. Akiyoshi
中科院分区：
文献类型：
--
作者：
Y. Iwasaki;Utae Takami;S. Sawada;K. Akiyoshi
用于减少非特异性生物污垢的表面设计是医疗器械制造中最重要的问题之一。本文提出了一种新合成的碳水化合物固定化磷酸胆碱聚合物，用于医疗器械的表面修饰，以控制与活细胞的界面。采用常规自由基聚合法制备了2-甲基丙烯酰氧乙基磷酸胆碱（MPC）、甲基丙烯酸正丁酯（BMA）和2-乳酸氨基甲基丙烯酸乙酯（LAMA）无规共聚物。共聚物中单体投料比调整为24/75/1 （MPC/BMA/LAMA）。共聚物（PMBL1.0）可以通过乙醇溶液的溶剂蒸发涂覆。将含asialal糖蛋白受体（ASGPRs）的人肝细胞肝癌细胞系（HepG2）细胞接种于PMBL1.0或聚BMA （PBMA）包被的PET板上。在PBMA上可见许多贴壁细胞，细胞分布良好，呈单层黏附。由于PMBL1.0细胞中含有ASGPRs，因此在PMBL1.0上观察到HepG2的粘附。PMBL1.0黏附的部分细胞呈球状形成，表面散布着类似形状的球体。培养96h后共聚焦激光显微镜观察发现，白蛋白的产生优先发生在球体的中心。粘附在PBMA上的细胞产生的白蛋白较少。ELISA法测定PMBL1.0粘附的细胞每单位白蛋白产量显著高于PBMA粘附的细胞。利用包被PMBL1.0的中空纤维微型模块进行HepG2的长期培养。HepG2产生的白蛋白浓度持续升高一个月。在迷你模块中，HepG2的功能在这段时间内得到了有效的保存。在中空纤维膜上也观察到HepG2细胞形成球形。综上所述，PMBL1.0可以为肝细胞的培养提供合适的表面，具有生产可靠的生物人工肝（BAL）装置的巨大潜力。
The surface design used for reducing nonspecific biofouling is one of the most important issues for the fabrication of medical devices. We present here a newly synthesized a carbohydrate-immobilized phosphorylcholine polymer for surface modification of medical devices to control the interface with living cells. A random copolymer composed of 2-methacryloyloxyethyl phosphorylcholine (MPC), n-butyl methacrylate (BMA), and 2-lactobionamidoethyl methacrylate (LAMA) was synthesized by conventional radical polymerization. The monomer feeding ratio in the copolymer was adjusted to 24/75/1 (MPC/BMA/LAMA). The copolymer (PMBL1.0) could be coated by solvent evaporation from an ethanol solution. Cells of the human hepatocellular liver carcinoma cell line (HepG2) having asialoglycoprotein receptors (ASGPRs) were seeded on PMBL1.0 or poly(BMA) (PBMA)-coated PET plates. On PBMA, many adherent cells were observed and were well spread with monolayer adhesion. HepG2 adhesion was observed on PMBL1.0 because the cell has ASGPRs. Furthermore, some of the cells adhering to PMBL1.0 had a spheroid formation and similarly shaped spheroids were scattered on the surface. According to confocal laser microscopic observation after 96h cultivation, it was found that albumin production preferentially occurred in the center of the spheroid. The albumin production of the cells that adhered to PBMA was sparse. The amount of albumin production per unit cell that adhered to PMBL1.0 was determined by ELISA and was significantly higher than that which adhered to PBMA. Long-term cultivation of HepG2 was also performed using hollow fiber mini-modules coated with PMBL1.0. The concentration of albumin produced from HepG2 increased continuously for one month. In the mini-module, the function of HepG2 was effectively preserved for that period. On the hollow fiber membrane, spheroid formation of HepG2 cells was also observed. In conclusion, PMBL1.0 can provide a suitable surface for the cultivation of hepatocytes and has great potential for producing reliable bioartificial liver (BAL) devices.