Induction of myofibroblastic differentiation in vitro by covalently immobilized transforming growth factor-beta(1).

Wolfgang Metzger, Nadine Grenner, Sandra E Motsch, Rothin Strehlow, Tim Pohlemann, Martin Oberringer
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引用次数: 19

Abstract

Growth factors are an important tool in tissue engineering. Bone morphogenetic protein-2 and transforming growth factor-beta(1) (TGF-beta(1)) are used to provide bioactivity to surgical implants and tissue substitute materials. Mostly growth factors are used in soluble or adsorbed form. However, simple adsorption of proteins to surfaces is always accompanied by reduced stability and undefined pharmacokinetics. This study aims to prove that TGF-beta(1) can be covalently immobilized to functionalized surfaces, maintaining its ability to induce myofibroblastic differentiation of normal human dermal fibroblasts. In vivo, fibroblasts differentiate to myofibroblasts (MFs) during soft tissue healing by the action of TGF-beta(1). As surfaces for our experiments, we used slides bearing aldehyde, epoxy, or amino groups. For our in vitro cell culture experiments, we used the expression of alpha-smooth muscle actin as a marker for MFs after immunochemical staining. Using the aldehyde and the epoxy slides, we were able to demonstrate the activity of immobilized TGF-beta(1) through a significant increase in MF differentiation rate. A simple immunological test was established to detect TGF-beta(1) on the surfaces. This technology enables the creation of molecular "landscapes" consisting of several factors arranged in a distinct spatial pattern and immobilized on appropriate surfaces.

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共价固定化转化生长因子- β诱导肌成纤维细胞的体外分化(1)。
生长因子是组织工程中的重要工具。骨形态发生蛋白-2和转化生长因子- β (1) (tgf - β(1))被用来为外科植入物和组织替代材料提供生物活性。大多数生长因子以可溶性或吸附形式使用。然而,蛋白质在表面的简单吸附总是伴随着稳定性降低和不确定的药代动力学。本研究旨在证明tgf - β(1)可以共价固定在功能化表面,维持其诱导正常人类真皮成纤维细胞向肌成纤维细胞分化的能力。在体内,在软组织愈合过程中,成纤维细胞在tgf - β的作用下分化为肌成纤维细胞(MFs)(1)。作为我们实验的表面,我们使用了含有醛、环氧或氨基的载玻片。在体外细胞培养实验中,我们使用α -平滑肌肌动蛋白的表达作为免疫化学染色后MFs的标记物。使用醛和环氧树脂载玻片,我们能够通过显著增加MF分化率来证明固定化tgf - β(1)的活性。建立了一种简单的免疫试验来检测表面上的tgf - β(1)。这种技术可以创造分子“景观”,由几个因素组成,以不同的空间模式排列,并固定在适当的表面上。
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Tissue engineering
Tissue engineering CELL & TISSUE ENGINEERING-BIOTECHNOLOGY & APPLIED MICROBIOLOGY
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