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通过改变溶液环境来控制基于弹性蛋白的聚合物涂层的表面粗糙度。

Manipulating the solution environment to control the surface roughness of elastin-based polymer coatings.

机构信息

Department of Biomedical Materials Science, 21693University of Mississippi Medical Center, Jackson, MS, USA.

出版信息

J Biomater Appl. 2021 Sep;36(3):419-427. doi: 10.1177/08853282211010302. Epub 2021 Apr 17.

Abstract

Elastin-like polypeptides (ELP) have been used as a genetically-engineered, biocompatible substitute for elastin. Cell culture coatings prepared using ELP conjugated to low molecular weight polyethyleneimine (PEI) entices cells to form three-dimensional cellular aggregates that mimic their counterparts. This study seeks to control the deposition of the ELP and ELP-PEI molecules to control the roughness of the final coatings. The two polymers were coated onto three different substrates (glass, polystyrene, tissue-culture polystyrene) and the solution environment was altered by changing the polymer concentration (0.5, 1.0, 1.5 mg/mL) and/or salt concentration (None, 0.2 M phosphate buffered saline) for a total of 36 conditions. Atomic force microscopy (AFM) was used to measure the average roughness (R) of the samples and found that ELP coated samples had a higher R than their ELP-PEI counterparts. The coatings were tested for stability by performing cell culture media changes every three days for 11 days. AFM showed that the average roughness of the tested samples increased with each media change. To address this, the surfaces were crosslinked using hexamethyl diisocyanate, which minimized the change in surface roughness even when subjected to an intense sonication process. This study provides parameters to achieve elastin-based coatings with controlled roughness that can be used to support stable, long-term cell culture.

摘要

弹性蛋白样多肽 (ELP) 已被用作一种经过基因工程改造的、生物相容性的弹性蛋白替代品。使用与低分子量聚乙烯亚胺 (PEI) 偶联的 ELP 制备的细胞培养涂层,诱使细胞形成三维细胞聚集体,从而模拟其对应物。本研究旨在控制 ELP 和 ELP-PEI 分子的沉积,以控制最终涂层的粗糙度。将这两种聚合物涂覆在三种不同的基底(玻璃、聚苯乙烯、组织培养聚苯乙烯)上,并通过改变聚合物浓度(0.5、1.0、1.5mg/mL)和/或盐浓度(无、0.2M 磷酸盐缓冲盐水)来改变溶液环境,共进行了 36 种条件的实验。原子力显微镜 (AFM) 用于测量样品的平均粗糙度 (R),发现 ELP 涂层样品的 R 值高于其 ELP-PEI 对应物。通过每三天更换一次细胞培养基,共进行 11 天,对涂层的稳定性进行测试。AFM 显示,随着每次培养基的变化,测试样品的平均粗糙度都有所增加。为了解决这个问题,使用六亚甲基二异氰酸酯对表面进行交联,即使在经受强烈的超声处理过程时,也能将表面粗糙度的变化最小化。本研究提供了实现具有可控粗糙度的基于弹性蛋白的涂层的参数,该涂层可用于支持稳定、长期的细胞培养。

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