Hu Shaohua, Eberhard Luc, Chen Jianxin, Love J Christopher, Butler James P, Fredberg Jeffrey J, Whitesides George M, Wang Ning
Physiology Program, Harvard School of Public Health, Boston, MA 02115, USA.
Am J Physiol Cell Physiol. 2004 Nov;287(5):C1184-91. doi: 10.1152/ajpcell.00224.2004. Epub 2004 Jun 22.
We describe a three-dimensional magnetic twisting device that is useful in characterizing the mechanical properties of cells. With the use of three pairs of orthogonally aligned coils, oscillatory mechanical torque was applied to magnetic beads about any chosen axis. Frequencies up to 1 kHz could be attained. Cell deformation was measured in response to torque applied via an RGD-coated, surface-bound magnetic bead. In both unpatterned and micropatterned elongated cells on extracellular matrix, the mechanical stiffness transverse to the long axis of the cell was less than half that parallel to the long axis. Elongated cells on poly-L-lysine lost stress fibers and exhibited little mechanical anisotropy; disrupting the actin cytoskeleton or decreasing cytoskeletal tension substantially decreased the anisotropy. These results suggest that mechanical anisotropy originates from intrinsic cytoskeletal tension within the stress fibers. Deformation patterns of the cytoskeleton and the nucleolus were sensitive to loading direction, suggesting anisotropic mechanical signaling. This technology may be useful for elucidating the structural basis of mechanotransduction.
我们描述了一种三维磁扭转装置,它可用于表征细胞的力学特性。通过使用三对正交排列的线圈,可围绕任何选定轴对磁珠施加振荡机械扭矩。可实现高达1kHz的频率。通过经RGD包被、表面结合的磁珠施加扭矩,测量细胞变形情况。在细胞外基质上的无图案和微图案化伸长细胞中,横向于细胞长轴的机械刚度小于平行于长轴的机械刚度的一半。聚-L-赖氨酸上的伸长细胞失去应力纤维,几乎没有表现出机械各向异性;破坏肌动蛋白细胞骨架或降低细胞骨架张力会显著降低各向异性。这些结果表明,机械各向异性源自应力纤维内的内在细胞骨架张力。细胞骨架和核仁的变形模式对加载方向敏感,表明存在各向异性机械信号传导。这项技术可能有助于阐明机械转导的结构基础。