The force loading rate drives cell mechanosensing through both reinforcement and cytoskeletal softening

Cell response to force regulates essential processes in health and disease. However, the fundamental mechanical variables that cells sense and respond to remain unclear. Here we show that the rate of force application (loading rate) drives mechanosensing, as predicted by a molecular clutch model. By applying dynamic force regimes to cells through substrate stretching, optical tweezers, and atomic force microscopy, we find that increasing loading rates trigger talin-dependent mechanosensing, leading to adhesion growth and reinforcement, and YAP nuclear localization. However, above a given threshold the actin cytoskeleton softens, decreasing loading rates and preventing reinforcement. By stretching rat lungs in vivo, we show that a similar phenomenon may occur. Our results show that cell sensing of external forces and of passive mechanical parameters (like tissue stiffness) can be understood through the same mechanisms, driven by the properties under force of the mechanosensing molecules involved.

The force loading rate drives cell mechanosensing through both reinforcement and fluidization

Cell response to force regulates essential processes in health and disease. However, the fundamental mechanical variables that cells sense and respond to remain unclear. Here we show that the rate of force application (loading rate) drives mechanosensing, as predicted by a molecular clutch model. By applying dynamic force regimes to cells through substrate stretching, optical tweezers, and atomic force microscopy, we find that increasing loading rates trigger talin-dependent mechanosensing, leading to adhesion growth and reinforcement, and YAP nuclear localization. However, above a given threshold the actin cytoskeleton undergoes fluidization and softens, decreasing loading rates and preventing reinforcement. By stretching rat lungs in vivo, we show that a similar phenomenon occurs at the organ level. Our results show that cell sensing of external forces and of passive mechanical parameters (like tissue stiffness) can be understood through the same mechanisms, driven by the properties under force of the mechanosensing molecules involved.

A Comprehensive Method for Accurate Strain Distribution Measurement of Cell Substrate Subjected to Large Deformation

Cell mechanical stretching in vitro is a fundamental technique commonly used in cardiovascular mechanobiology research. Accordingly, it is crucial to measure the accurate strain field of cell substrate under different strains. Digital image correlation (DIC) is a widely used measurement technique, which is able to obtain the accurate displacement and strain distribution. However, the traditional DIC algorithm used in digital image correlation engine (DICe) cannot obtain accurate result when utilized in large strain measurement. In this paper, an improved method aiming to acquire accurate strain distribution of substrate in large deformation was proposed, to evaluate the effect and accuracy, based on numerical experiments. The results showed that this method was effective and highly accurate. Then, we carried out uniaxial substrate stretching experiments and applied our method to measure strain distribution of the substrate. The proposed method could obtain accurate strain distribution of substrate film during large stretching, which would allow researchers to adequately describe the response of cells to different strains of substrate.

Numerical Simulation about Stretching Process in Different Layers of Cartilage

Cartilage with complex structure is a porous viscoelastic material. The direction of arrangement of collagen fibers in different layer regions directly affects the mechanical properties of the cartilage layer region. It is very important to use the method of numerical simulation for studying cartilage damage and repair through experimental measurements of cartilage mechanical parameters of the different layers. Because of the relatively small size of the cartilage, it is very difficult to measure mechanical parameters of cartilages by tensile test. The paper for main problems in the tensile test of cartilages, first by porcine articular cartilage compression testing, measuring the displacement of cartilage areas of different layers, according to the characteristics of the displacement determines the size of areas of different layers of cartilage, and then designed the cartilage and substrate stretching models. Model includes two forms of direct bonding and embedding bonding to simulate stretching process of different layers of the cartilage area in numerical way, displacement fields and stress-strain fields of stretching cartilage in different layer regions are derived. The numerical results show that using the way of embedded bonding can make stress of articular well-distributed without stress concentration, so it is a good way of bonding methods. Paper of the research work laid the foundation for measuring mechanical parameters of cartilage by stretch experiment.

Reproducible uniform equibiaxial stretch of precision-cut lung slices

Simulating ventilator-induced lung injury (VILI) in the laboratory requires stretching of lung alveolar tissue. Whereas precision-cut lung slices (PCLSs) are widely used for studying paracrine signaling pathways in the lungs, their use in stretch studies is very limited because of the technical challenge of fixing them to a stretchable substrate, stretching them uniformly, or holding them in a stretch device without causing rupture. We describe a novel method for attaching PCLSs to silicone membranes by stitching them together in a star-shaped pattern. Using a device that was previously designed in our laboratory for stretching primary alveolar epithelial cell monolayers, we demonstrate that in the central region of the PCLSs stretch is uniform, equibiaxial, and, after a short preconditioning period, also reproducible. The stitched and stretched PCLSs showed equal or better viability outcomes after 60 min of cyclic stretch at different magnitudes of physiological stretch compared with primary pulmonary alveolar epithelial cell monolayers. Preparing and stitching the PCLSs before stretch is relatively easy to perform, yields repeatable outcomes, and can be used with tissue from any species. Together with the ensuring uniform and equibiaxial stretch, the proposed methods provide an optimal model for VILI studies with PCLSs.