Differential integrin expression regulates cell sensing of the matrix nanoscale geometry
Differential integrin expression regulates cell sensing of the matrix nanoscale geometry
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DOI：
10.1016/j.actbio.2016.11.069
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发表时间：
2017-03-01
影响因子：
9.7
通讯作者：
Gautrot, Julien E.
中科院分区：
文献类型：
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作者：
Di Cio, Stefania;Boggild, Thea M. L.;Gautrot, Julien E.
细胞外基质（ECM）的纳米级几何形状和形貌是控制细胞粘附和表型的重要参数。类似地，整合素表达和它们调节的粘附的几何成熟与细胞铺展和表型的重要变化相关。然而，整合素表达如何控制ECM几何形状的纳米级感测尚不清楚。在这里，我们开发了一种新的纳米图案化技术，静电纺丝光刻（ENL），它允许生产具有受控尺寸（250-1000 nm）和密度的准二维纤维纳米图案。ENL依靠电纺纤维作为抗蛋白质聚合物刷受控生长的掩模。使用SEM、AFM和免疫荧光成像来观察所得图案和细胞外基质蛋白纤连蛋白对图案化纤维的吸附。研究了粘附形成的控制，以及新型基质的重塑和沉积。发现细胞扩散受纤维大小的调节，类似于先前对圆形纳米片的观察。然而，细胞形状和极性受到更显著的影响。这些变化与重要的细胞骨架重组，随着图案尺寸的减少，应力纤维的形成逐渐减少。最后，发现工程化细胞系中ccv 03和α 5131整联蛋白的差异表达是细胞感知ECM的纳米级几何形状的重要介质。重要性声明本研究中开发的新的EBM模式，通过ENL，模拟组织基质中发现的天然基质的几何形状和连续性，同时保持准2D特性（以便于成像和与其它2D系统例如由胶体光刻产生的微图案化单层和圆形纳米片进行比较）。这些结果表明，ECM的纳米级几何形状在调节细胞粘附中起着重要作用，并且这是由整合素表达调节的。这是一个重要的发现，因为它意味着整合素介导的特定细胞类型的粘附机制的生化背景的知识，应该允许更好的设计生物材料和生物界面。事实上，整联蛋白表达的变化通常与细胞增殖和分化的控制有关。(C)2016 Acta Materialia Inc.由爱思唯尔有限公司出版。保留所有权利。
The nanoscale geometry and topography of the extra-cellular matrix (ECM) is an important parameter controlling cell adhesion and phenotype. Similarly, integrin expression and the geometrical maturation of adhesions they regulate have been correlated with important changes in cell spreading and phenotype. However, how integrin expression controls the nanoscale sensing of the ECM geometry is not clearly understood. Here we develop a new nanopatterning technique, electrospun nanofiber lithography (ENL), which allows the production of a quasi-2D fibrous nanopattern with controlled dimensions (250-1000 nm) and densities. ENL relies on electrospun fibres to act as a mask for the controlled growth of protein-resistant polymer brushes. SEM, AFM and immunofluorescence imaging were used to characterise the resulting patterns and the adsorption of the extra-cellular matrix protein fibronectin to the patterned fibres. The control of adhesion formation was studied, as well as the remodelling and deposition of novel matrix. Cell spreading was found to be regulated by the size of fibres, similarly to previous observations made on circular nanopatterns. However, cell shape and polarity were more significantly affected. These changes correlated with important cytoskeleton reorganisation, with a gradual decrease in stress fibre formation as the pattern dimensions decrease. Finally, the differential expression of ccv03 and ot5131 integrins in engineered cell lines was found to be an important mediator of cell sensing of the nanoscale geometry of the ECM.Statement of SignificanceThe novel nanofiber patterns developed in this study, via ENL, mimic the geometry and continuity of natural matrices found in the stroma of tissues, whilst preserving a quasi-2D character (to facilitate imaging and for comparison with other 2D systems such as micropatterned monolayers and circular nanopatches generated by colloidal lithography). These results demonstrate that the nanoscale geometry of the ECM plays an important role in regulating cell adhesion and that this is modulated by integrin expression. This is an important finding as it implies that the knowledge of the biochemical context underlying the integrin-mediated adhesive machinery of specific cell types should allow better design of biomaterials and biointerfaces. Indeed, changes in integrin expression are often associated with the control of cell proliferation and differentiation. (C) 2016 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.