Nano-mechanical mapping of interdependent cell and ECM mechanics by AFM force spectroscopy
Nano-mechanical mapping of interdependent cell and ECM mechanics by AFM force spectroscopy
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DOI：
10.1038/s41598-019-48566-7
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发表时间：
2019-08-23
影响因子：
4.6
通讯作者：
Radmacher, Manfred
中科院分区：
文献类型：
--
作者：
Babu, Prem Kumar Viji;Rianna, Carmela;Radmacher, Manfred
细胞外基质（ECM）作为组织的动态成分，影响细胞行为，并在细胞力学和组织稳态中发挥重要作用。相反，这种三维支架会被细胞动态、结构和机械地修改。在生物物理学领域，细胞和ECM力学的独立作用已得到广泛研究；然而，缺乏同时测量的细胞和 ECM 力学之间相互依赖的相互作用的实验数据。在这里，我们使用原子力显微镜 (AFM) 表征了五种不同的脱细胞基质，它们的形貌、ECM 成分和硬度各不相同，并将它们与正常和病理性成纤维细胞（疤痕和掌腱膜成纤维细胞）一起培养。我们分别使用 AFM 峰值力成像和机械测绘研究了由于细胞接种而导致的这些基质的形貌和弹性的变化。我们发现正常成纤维细胞比病理成纤维细胞更能软化这些基质，这表明病理成纤维细胞深刻影响纤维化过程中的组织硬化。我们检测到此处使用的脱细胞基质的不同 ECM 成分会影响成纤维细胞硬度，从而强调细胞力学不仅取决于 ECM 硬度，还取决于其成分。我们使用共聚焦显微镜来评估成纤维细胞的侵袭，发现病理成纤维细胞比正常成纤维细胞侵入基质更深。
Extracellular matrix (ECM), as a dynamic component of the tissue, influences cell behavior and plays an important role in cell mechanics and tissue homeostasis. Reciprocally, this three-dimensional scaffold is dynamically, structurally and mechanically modified by cells. In the field of biophysics, the independent role of cell and ECM mechanics has been largely investigated; however, there is a lack of experimental data reporting the interdependent interplay between cell and ECM mechanics, measured simultaneously. Here, using Atomic Force Microscopy (AFM) we have characterized five different decellularized matrices diverse in their topography, ECM composition and stiffness and cultured them with normal and pathological fibroblasts (scar and Dupuytren's). We investigated the change in topography and elasticity of these matrices due to cell seeding, by using AFM peak force imaging and mechanical mapping, respectively. We found normal fibroblasts soften these matrices more than pathological fibroblasts, suggesting that pathological fibroblasts are profoundly influencing tissue stiffening in fibrosis. We detected different ECM composition of decellularized matrices used here influences fibroblast stiffness, thus highlighting that cell mechanics not only depends on ECM stiffness but also on their composition. We used confocal microscopy to assess fibroblasts invasion and found pathological fibroblasts were invading the matrices deeper than normal fibroblasts.