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用于新型单轴拉伸应变生物反应器的弹性细胞培养底物的开发。

Development of an elastic cell culture substrate for a novel uniaxial tensile strain bioreactor.

作者信息

Moles Matthew D, Scotchford Colin A, Ritchie Alastair Campbell

机构信息

Division of Materials, Mechanics and Structures, Faculty of Mechanical, Materials and Manufacturing Engineering, University of Nottingham, University Park, Nottingham, NG7 2RD, United Kingdom.

出版信息

J Biomed Mater Res A. 2014 Jul;102(7):2356-64. doi: 10.1002/jbm.a.34917. Epub 2013 Aug 31.

Abstract

Bioreactors can be used for mechanical conditioning and to investigate the mechanobiology of cells in vitro. In this study a polyurethane (PU), Chronoflex AL, was evaluated for use as a flexible cell culture substrate in a novel bioreactor capable of imparting cyclic uniaxial tensile strain to cells. PU membranes were plasma etched, across a range of operating parameters, in oxygen. Contact angle analysis and X-ray photoelectron spectroscopy showed increases in wettability and surface oxygen were related to both etching power and duration. Atomic force microscopy demonstrated that surface roughness decreased after etching at 20 W but was increased at higher powers. The etching parameters, 20 W 40 s, produced membranes with high surface oxygen content (21%), a contact angle of 66° ± 7° and reduced topographical features. Etching and protein conditioning membranes facilitated attachment, and growth to confluence within 3 days, of MG-63 osteoblasts. After 2 days with uniaxial strain (1%, 30 cycles/min, 1500 cycles/day), cellular alignment was observed perpendicular to the principal strain axis, and found to increase after 24 h. The results indicate that the membrane supports culture and strain transmission to adhered cells.

摘要

生物反应器可用于机械调节并在体外研究细胞的力学生物学。在本研究中,对一种聚氨酯(PU)材料Chronoflex AL进行了评估,以用作一种新型生物反应器中的柔性细胞培养底物,该生物反应器能够向细胞施加周期性单轴拉伸应变。在一系列操作参数下,PU膜在氧气中进行了等离子体蚀刻。接触角分析和X射线光电子能谱表明,润湿性的增加和表面氧含量与蚀刻功率和持续时间均有关。原子力显微镜显示,在20W功率下蚀刻后表面粗糙度降低,但在更高功率下则增加。蚀刻参数20W 40s产生的膜具有高表面氧含量(21%)、66°±7°的接触角以及减少的形貌特征。蚀刻和蛋白质处理的膜促进了MG-63成骨细胞的附着,并在三天内生长至汇合。在单轴应变(1%,30次循环/分钟,1500次循环/天)处理两天后,观察到细胞垂直于主应变轴排列,并发现24小时后排列增加。结果表明,该膜支持细胞培养并能将应变传递给贴壁细胞。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/65e8/4255296/b04995bcb68a/jbma0102-2356-f1.jpg

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