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表面化学在细胞-材料界面蛋白质重排中的作用。

Role of surface chemistry in protein remodeling at the cell-material interface.

机构信息

Center for Biomaterials and Tissue Engineering, Universidad Politécnica de Valencia, Valencia, Spain.

出版信息

PLoS One. 2011 May 9;6(5):e19610. doi: 10.1371/journal.pone.0019610.

Abstract

BACKGROUND

The cell-material interaction is a complex bi-directional and dynamic process that mimics to a certain extent the natural interactions of cells with the extracellular matrix. Cells tend to adhere and rearrange adsorbed extracellular matrix (ECM) proteins on the material surface in a fibril-like pattern. Afterwards, the ECM undergoes proteolytic degradation, which is a mechanism for the removal of the excess ECM usually approximated with remodeling. ECM remodeling is a dynamic process that consists of two opposite events: assembly and degradation.

METHODOLOGY/PRINCIPAL FINDINGS: This work investigates matrix protein dynamics on mixed self-assembled monolayers (SAMs) of -OH and -CH(3) terminated alkanethiols. SAMs assembled on gold are highly ordered organic surfaces able to provide different chemical functionalities and well-controlled surface properties. Fibronectin (FN) was adsorbed on the different surfaces and quantified in terms of the adsorbed surface density, distribution and conformation. Initial cell adhesion and signaling on FN-coated SAMs were characterized via the formation of focal adhesions, integrin expression and phosphorylation of FAKs. Afterwards, the reorganization and secretion of FN was assessed. Finally, matrix degradation was followed via the expression of matrix metalloproteinases MMP2 and MMP9 and correlated with Runx2 levels. We show that matrix degradation at the cell material interface depends on surface chemistry in MMP-dependent way.

CONCLUSIONS/SIGNIFICANCE: This work provides a broad overview of matrix remodeling at the cell-material interface, establishing correlations between surface chemistry, FN adsorption, cell adhesion and signaling, matrix reorganization and degradation. The reported findings improve our understanding of the role of surface chemistry as a key parameter in the design of new biomaterials. It demonstrates the ability of surface chemistry to direct proteolytic routes at the cell-material interface, which gains a distinct bioengineering interest as a new tool to trigger matrix degradation in different biomedical applications.

摘要

背景

细胞-材料相互作用是一个复杂的双向动态过程,在某种程度上模拟了细胞与细胞外基质的自然相互作用。细胞倾向于在材料表面以纤维状模式附着和重排吸附的细胞外基质(ECM)蛋白。随后,ECM 经历蛋白水解降解,这是一种去除多余 ECM 的机制,通常近似于重塑。ECM 重塑是一个动态过程,由两个相反的事件组成:组装和降解。

方法/主要发现:本工作研究了混合自组装单层(SAM)中基质蛋白的动力学,该混合自组装单层由-OH 和-CH(3)终止的烷硫醇组成。在金上组装的 SAM 是高度有序的有机表面,能够提供不同的化学功能和可控制的表面特性。纤连蛋白(FN)被吸附在不同的表面上,并根据吸附表面密度、分布和构象进行定量。通过形成粘着斑、整合素表达和粘着斑激酶(FAKs)磷酸化,对 FN 涂层 SAM 上初始细胞黏附和信号转导进行了表征。随后,评估了 FN 的重排和分泌。最后,通过基质金属蛋白酶 MMP2 和 MMP9 的表达以及与 Runx2 水平的相关性,来跟踪基质降解。我们表明,细胞-材料界面处的基质降解取决于 MMP 依赖的表面化学。

结论/意义:本工作提供了细胞-材料界面处基质重塑的广泛概述,建立了表面化学、FN 吸附、细胞黏附和信号转导、基质重排和降解之间的相关性。所报道的发现提高了我们对表面化学作为新生物材料设计关键参数的作用的理解。它展示了表面化学在细胞-材料界面处引导蛋白水解途径的能力,这作为一种在不同的生物医学应用中触发基质降解的新工具,具有独特的生物工程意义。

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