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用于评估聚合物性质对原代人角质形成细胞和纤维原细胞在单独和共培养中的影响的测试系统。

Test system for evaluating the influence of polymer properties on primary human keratinocytes and fibroblasts in mono- and coculture.

机构信息

Institute of Biomaterial Science and Berlin-Brandenburg Center for Regenerative Therapies, Helmholtz-Zentrum Geesthacht, Kantstrasse 55, 14513 Teltow, Germany.

出版信息

J Biotechnol. 2013 Jun 20;166(1-2):58-64. doi: 10.1016/j.jbiotec.2013.04.012. Epub 2013 Apr 30.

Abstract

Interactions of cells with polymer-based biomaterials are influenced by properties of the substrate. Polymers, which are able to induce cell specific effects, gain increasing importance for biotechnology and regenerative therapies. A test system was developed, which allows studying primary human keratinocytes and fibroblasts in mono- and cocultures to analyze and operate the effect of polymer properties. This system offers to identify polymers for keratinocyte cultivation or wound dressings, since adherence, viability and functionality can be analyzed. Especially the coculture system enables the characterization of potential cell specific effects of polymer-based biomaterials. To establish a coculture test system, it is challenging to find a suitable culture medium, to identify initial seeding densities for comparable cell growth and to develop methods to distinguish and characterize both cell types. Poly(n-butyl acrylate) networks (cPnBAs) as model biomaterials were used to demonstrate the applicability of our newly developed coculture screening system for differential cell growth. The apparent Young's modulus of the cPnBAs differentially regulated fibroblasts and keratinocytes. Particularly, cPnBA73 with an apparent Young's modulus of 930±140 kPa measured in phosphate buffered saline (PBS) solution at ambient temperature seemed to have favoring properties for keratinocyte adhesion, while fibroblast adhesion was not affected. For keratinocytes the concentration of some pro-inflammatory cytokines was lower on cPnBA73 and a decreased deposition of collagen, elastin and fibronectin was observed in the coculture.

摘要

细胞与基于聚合物的生物材料的相互作用受基底性质的影响。能够诱导细胞特异性效应的聚合物在生物技术和再生疗法中越来越重要。开发了一种测试系统,该系统允许研究原代人角质形成细胞和成纤维细胞的单培养和共培养,以分析和操作聚合物性质的影响。该系统可用于鉴定用于角质形成细胞培养或伤口敷料的聚合物,因为可以分析粘附性、活力和功能。特别是共培养系统能够表征基于聚合物的生物材料的潜在细胞特异性效应。为了建立共培养测试系统,找到合适的培养基、确定用于可比细胞生长的初始接种密度以及开发区分和表征两种细胞类型的方法都是具有挑战性的。聚(正丁基丙烯酸酯)网络(cPnBAs)作为模型生物材料被用于证明我们新开发的共培养筛选系统对差异细胞生长的适用性。cPnBAs 的表观杨氏模量差异调节成纤维细胞和角质形成细胞。特别是,在环境温度下磷酸盐缓冲盐水(PBS)溶液中测量的表观杨氏模量为 930±140 kPa 的 cPnBA73 似乎具有有利于角质形成细胞粘附的性质,而成纤维细胞粘附不受影响。对于角质形成细胞,cPnBA73 上某些促炎细胞因子的浓度较低,并且在共培养中观察到胶原蛋白、弹性蛋白和纤维连接蛋白的沉积减少。

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