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一种用于原位研究细胞和组织在机械应变下的新平台。

A novel platform for in situ investigation of cells and tissues under mechanical strain.

机构信息

Department of Mechanical Sciences & Engineering, University of Illinois at Urbana-Champaign, 1206 W. Green St., Urbana, IL 61801, USA.

出版信息

Acta Biomater. 2010 Aug;6(8):2979-90. doi: 10.1016/j.actbio.2010.02.035. Epub 2010 Feb 25.

Abstract

The mechanical micro-environment influences cellular responses such as migration, proliferation, differentiation and apoptosis. Cells are subjected to mechanical stretching in vivo, e.g., epithelial cells during embryogenesis. Current methodologies do not allow high-resolution in situ observation of cells and tissues under applied strain, which may reveal intracellular dynamics and the origin of cell mechanosensitivity. A novel polydimethylsiloxane substrate was developed, capable of applying tensile and compressive strain (up to 45%) to cells and tissues while allowing in situ observation with high-resolution optics. The strain field of the substrate was characterized experimentally using digital image correlation, and the deformation was modeled by the finite element method, using a Mooney-Rivlin hyperelastic constitutive relation. The substrate strain was found to be uniform for >95% of the substrate area. As a demonstration of the system, mechanical strain was applied to single fibroblasts transfected with GFP-actin and whole transgenic Drosophila embryos expressing GFP in all neurons during live imaging. Three observations of biological responses due to applied strain are reported: (1) dynamic rotation of intact actin stress fibers in fibroblasts; (2) lamellipodia activity and actin polymerization in fibroblasts; (3) active axonal contraction in Drosophila embryo motor neurons. The novel platform may serve as an important tool in studying the mechanoresponse of cells and tissues, including whole embryos.

摘要

机械微环境影响细胞的反应,如迁移、增殖、分化和凋亡。细胞在体内受到机械拉伸,例如胚胎发生过程中的上皮细胞。目前的方法学无法在施加应变下对细胞和组织进行高分辨率的原位观察,这可能揭示细胞内动力学和细胞机械敏感性的起源。开发了一种新型的聚二甲基硅氧烷(PDMS)基底,能够对细胞和组织施加拉伸和压缩应变(高达 45%),同时允许使用高分辨率光学进行原位观察。通过数字图像相关实验对基底的应变场进行了表征,并使用有限元法对变形进行建模,使用 Mooney-Rivlin 超弹性本构关系。发现基底应变在>95%的基底面积上是均匀的。作为系统的演示,在活细胞成像中,对转染 GFP-肌动蛋白的单个成纤维细胞和表达 GFP 的整个转基因果蝇胚胎施加机械应变。报道了由于施加应变而产生的三个生物学反应观察结果:(1)成纤维细胞中完整的肌动蛋白应力纤维的动态旋转;(2)成纤维细胞中的片状伪足活性和肌动蛋白聚合;(3)果蝇胚胎运动神经元中的活跃轴突收缩。该新型平台可用作研究细胞和组织机械反应的重要工具,包括整个胚胎。

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