Rapid hepatic cell attachment onto biodegradable polymer surfaces without toxicity using an avidin-biotin binding system
Rapid hepatic cell attachment onto biodegradable polymer surfaces without toxicity using an avidin-biotin binding system
复制标题
DOI：
10.1016/j.biomaterials.2006.05.026
复制
发表时间：
2006-10-01
期刊：
影响因子：
14
通讯作者：
Sakai, Yasuyuki
中科院分区：
文献类型：
--
作者：
Kojima, Nobuhiko;Matsuo, Tomoki;Sakai, Yasuyuki
细胞在生物可降解高分子支架上的有效附着是组织工程的必要前提。然而，很难用细胞均匀地覆盖支架表面，因为支架通常是高度多孔的，具有复杂的三维（3D）表面。在这篇文章中，我们证明了亲和素-生物素结合系统（ABBS）的生物素化的Hep G2细胞的初始附着的亲和素吸附的平面，二维（2D）和高度多孔的3D聚L-乳酸（PLLA）表面的效率。还研究了生物素化和/或强ABBS结合力的潜在毒性。ABBS辅助Hep G2细胞在10分钟内粘附到平坦的PLLA表面;这些附着的细胞的增殖与在胶原上培养的对照完整细胞相当。附着细胞的肝功能（如白蛋白分泌、CYP 1A 1和CYP 1A 2基因诱导以及通过乙氧基试卤灵O-脱乙基酶试验测定的CYP 1A 1/2代谢能力）未发生显著变化。此外，细胞因子的刺激：制瘤素M（OSM）磷酸化的细胞内信号分子，细胞外信号相关激酶I（ERK 1）通过跨膜受体复合物，在接种ABBS后24小时。此外，证明了Hep G2细胞与高度多孔的PLLA 3D支架的有效附着。这些结果清楚地表明，ABBS可用于在可生物降解的、基于聚合物的平坦2D表面和高度多孔的3D支架中快速捕获细胞。此外，通过这种技术结合肝细胞对肝脏特异性功能或跨膜受体复合物的信号传递能力的影响很小。(c)2006爱思唯尔有限公司版权所有。
Efficient cell attachment to biodegradable polymer scaffolds is a necessary prerequisite in tissue engineering. However, it is difficult to evenly cover scaffold surfaces with cells because scaffolds are generally highly porous, with complex three-dimensional (3D) surfaces. In this article, we demonstrate the efficiency of avidin-biotin binding systems (ABBS) for the initial attachment of biotinylated Hep G2 cells to avidin adsorbed flat, two-dimensional (2D) and highly porous 3D Poly L-lactic acid (PLLA) surfaces. The potential toxicity of biotinylation and/or strong ABBS binding forces was also investigated. ABBS assisted Hep G2 cells to adhere to a flat PLLA surface within 10 min; the proliferation of these attached cells was comparable with control intact cells cultured on collagen. Hepatic functions of the attached cells, such as albumin secretion, induction of CYP1A1 and CYP1A2 genes, and metabolic capacity of CYP1A1/2 as measured by the ethoxyresorufin O-deethylase assay, were not significantly changed. Also, a stimulus of a cytokine: oncostatin M (OSM) phosphorylated an intracellular signaling molecule, extracellular signal-related kinase I (ERK1) via transmembrane receptor complex, at 24 h after inoculation by ABBS. In addition, efficient attachment of Hep G2 cells to a highly porous PLLA 3D scaffold was demonstrated. These results clearly show that ABBS is useful for rapidly trapping cells in both biodegradable, polymer-based, flat 2D surfaces, and in highly porous 3D scaffolds. Furthermore, binding hepatic cells by this technique has only small effects on liver-specific functions, or on signal transfer ability of transmembrane receptor complexes. (c) 2006 Elsevier Ltd. All rights reserved.