Codan B, Del Favero G, Martinelli V, Long C S, Mestroni L, Sbaizero O
Department of Engineering and Architecture, University of Trieste, Italy.
Department of Engineering and Architecture, University of Trieste, Italy; International Center for Genetic Engineering and Biotechnology, Trieste, Italy.
Mater Sci Eng C Mater Biol Appl. 2014 Jul 1;40:427-34. doi: 10.1016/j.msec.2014.04.003. Epub 2014 Apr 13.
AFM was used to collect the whole force-deformation cell curves. They provide both the elasticity and adhesion behavior of mouse primary cardiac fibroblasts. To confirm the hypothesis that a link exists between the membrane receptors and the cytoskeletal filaments causing therefore changing in both elasticity and adhesion behavior, actin-destabilizing Cytochalsin D was administrated to the fibroblasts. From immunofluorescence observation and AFM loading/unloading curves, cytoskeletal reorganization as well as a change in the elasticity and adhesion was indeed observed. Elasticity of control fibroblasts is three times higher than that for fibroblasts treated with 0.5 μM Cytochalasin. Moreover, AFM loading-unloading curves clearly show the different mechanical behavior of the two different cells analyzed: (i) for control cells the AFM cantilever rises during the dwell time while cells with Cytochalasin fail to show such an active resistance; (ii) the maximum force to deform control cells is quite higher and as far as adhesion is concern (iii) the maximum separation force, detachment area and the detachment process time are much larger for control compared to the Cytochalasin treated cells. Therefore, alterations in the cytoskeleton suggest that a link must exist between the membrane receptors and the cytoskeletal filaments beneath the cellular surface and inhibition of actin polymerization has effects on the whole cell mechanical behavior as well as adhesion.
原子力显微镜(AFM)用于收集整个细胞的力-变形曲线。这些曲线能反映小鼠原代心脏成纤维细胞的弹性和黏附行为。为了证实细胞膜受体与细胞骨架丝之间存在联系,进而导致弹性和黏附行为发生变化这一假设,向成纤维细胞中加入了破坏肌动蛋白的细胞松弛素D。通过免疫荧光观察和AFM加载/卸载曲线,确实观察到了细胞骨架的重组以及弹性和黏附的变化。对照成纤维细胞的弹性比用0.5μM细胞松弛素处理的成纤维细胞高两倍。此外,AFM加载-卸载曲线清楚地显示了所分析的两种不同细胞的不同力学行为:(i)对于对照细胞,AFM悬臂在停留时间内上升,而用细胞松弛素处理的细胞则没有显示出这种主动抗性;(ii)使对照细胞变形的最大力要高得多,就黏附而言(iii)与用细胞松弛素处理的细胞相比,对照细胞的最大分离力、分离面积和分离过程时间要大得多。因此,细胞骨架的改变表明细胞表面下的膜受体与细胞骨架丝之间必然存在联系,并且肌动蛋白聚合的抑制会对整个细胞的力学行为以及黏附产生影响。