Sbaizero O, DelFavero G, Martinelli V, Long C S, Mestroni L
Department of Engineering and Architecture, University of Trieste, Italy; University of Colorado Cardiovascular Institute, University of Colorado Denver, Aurora, USA.
Department of Engineering and Architecture, University of Trieste, Italy.
Mater Sci Eng C Mater Biol Appl. 2015 Apr;49:217-224. doi: 10.1016/j.msec.2014.12.083. Epub 2014 Dec 27.
Atomic force microscopy (AFM) for single-cell force spectroscopy (SCFS) and Poisson statistic were used to analyze the detachment work recorded during the removal of gold-covered microspheres from cardiac fibroblasts. The effect of Cytochalasin D, a disruptor of the actin cytoskeleton, on cell adhesion was also tested. The adhesion work was assessed using a Poisson analysis also derived from single-cell force spectroscopy retracting curves. The use of Poisson analysis to get adhesion work from AFM curves is quite a novel method, and in this case, proved to be effective to study the short-range and long-range contributions to the adhesion work. This method avoids the difficult identification of minor peaks in the AFM retracting curves by creating what can be considered an average adhesion work. Even though the effect of actin depolymerisation is well documented, its use revealed that control cardiac fibroblasts (CT) exhibit a work of adhesion at least 5 times higher than that of the Cytochalasin treated cells. However, our results indicate that in both cells short-range and long-range contributions to the adhesion work are nearly equal and the same heterogeneity index describes both cells. Therefore, we infer that the different adhesion behaviors might be explained by the presence of fewer membrane adhesion molecules available at the AFM tip-cell interface under circumstances where the actin cytoskeleton has been disrupted.
利用用于单细胞力谱分析(SCFS)的原子力显微镜(AFM)和泊松统计分析从心脏成纤维细胞上移除金覆盖微球过程中记录的脱离功。还测试了肌动蛋白细胞骨架破坏剂细胞松弛素D对细胞粘附的影响。粘附功也使用同样源自单细胞力谱分析回缩曲线的泊松分析进行评估。使用泊松分析从AFM曲线获得粘附功是一种相当新颖的方法,在这种情况下,事实证明该方法对于研究粘附功的短程和长程贡献是有效的。该方法通过创建可被视为平均粘附功的量来避免在AFM回缩曲线中难以识别微小峰值的问题。尽管肌动蛋白解聚的作用已有充分记录,但它的使用表明对照心脏成纤维细胞(CT)的粘附功比用细胞松弛素处理的细胞至少高5倍。然而,我们的结果表明,在这两种细胞中,对粘附功的短程和长程贡献几乎相等,并且相同的异质性指数描述了这两种细胞。因此,我们推断,不同的粘附行为可能是由于在肌动蛋白细胞骨架被破坏的情况下,AFM针尖 - 细胞界面处可用的膜粘附分子较少所致。