Characterization of Cancer Cell Mechanics by Measuring Active Deformation Behavior.

Active deformation behavior reflects cell structural dynamics adapting to varying environmental constraints during malignancy progression. In most cases, cell mechanics is characterized by modeling using static equilibrium systems, which fails to comprehend cell deformation behavior leading to inaccuracies in distinguishing cancer cells from normal cells. Here, a method is introduced to measure the active deformation behavior of cancer cells using atomic force microscopy (AFM) and the newly developed deformation behavior cytometry (DBC). During the measurement, cells are deformed and allows a long timescale relaxation (≈5 s). Two parameters are derived to represent deformation behavior: apparent Poisson's ratio for adherent cells, which is measured with AFM and refers to the ratio of the lateral strain to the longitudinal strain of the cell, and shape recovery for suspended cells, which is measured with DBC. Active deformation behavior defines cancer cell mechanics better than traditional mechanical parameters (e.g., stiffness, diffusion, and viscosity). Additionally, aquaporins are essential for promoting the deformation behavior, while the actin cytoskeleton acts as a downstream effector. Therefore, the potential application of the cancer cell active deformation behavior as a biomechanical marker or therapeutic target in cancer treatment should be evaluated.

AFM-Based Poroelastic@Membrane Analysis of Cells and its Opportunities for Translational Medicine.

Cell mechanics is an emerging field of research for translational medicine. Here, the cell is modeled as poroelastic cytoplasm wrapped by tensile membrane (poroelastic@membrane model) and is characterized by the atomic force microscopy (AFM). The parameters of cytoskeleton network modulus EC , cytoplasmic apparent viscosity ηC , and cytoplasmic diffusion coefficient DC are used to describe the mechanical behavior of cytoplasm, and membrane tension γ is used to evaluate the cell membrane. Poroelastic@membrane analysis of breast cells and urothelial cells show that non-cancer cells and cancer cells have different distribution regions and distribution trends in the four-dimensional space composed of EC , ηC . From non-cancer to cancer cells, there is often a trend of γ, EC , ηC decreases and DC increases. Patients with urothelial carcinoma at different malignant stages can be distinguished at high sensitivity and specificity by analyzing the urothelial cells from tissue or urine. However, sampling directly from tumor tissues is an invasive method, may lead to undesirable consequences. Thus, AFM-based poroelastic@membrane analysis of urothelial cells from urine may provide a non-invasive and no-bio-label method to detecting urothelial carcinoma.

Cell senescence alters responses of porcine trabecular meshwork cells to shear stress.

Mechanical microenvironment and cellular senescence of trabecular meshwork cells (TMCs) are suspected to play a vital role in primary open-angle glaucoma pathogenesis. However, central questions remain about the effect of shear stress on TMCs and how aging affects this process. We have investigated the effect of shear stress on the biomechanical properties and extracellular matrix regulation of normal and senescent TMCs. We found a more significant promotion of Fctin formation, a more obvious realignment of F-actin fibers, and a more remarkable increase in the stiffness of normal cells in response to the shear stress, in comparison with that of senescent cells. Further, as compared to normal cells, senescent cells show a reduced extracellular matrix turnover after shear stress stimulation, which might be attributed to the different phosphorylation levels of the extracellular signal-regulated kinase. Our results suggest that TMCs are able to sense and respond to the shear stress and cellular senescence undermines the mechanobiological response, which may lead to progressive failure of cellular TM function with age.

AFM Force Relaxation Curve Reveals That the Decrease of Membrane Tension Is the Essential Reason for the Softening of Cancer Cells.

Differences in stiffness constitute an extremely important aspect of the mechanical differences between cancer cells and normal cells, and atomic force microscopy (AFM) is the most commonly used tool to characterize the difference in stiffness. However, the process of mechanical characterization using AFM has been controversial and the influence of the membrane tension on AFM measurement results was often ignored. Here, a physical model involving a simultaneous consideration of the effects of the cell membrane, cytoskeleton network and cytosol was proposed. We carried out a theoretical analysis of AFM force relaxation curves, and as a result solved many of the remaining controversial issues regarding AFM-based mechanical characterization of cells, and provided a quantitative solution for the membrane tension measured using AFM indentation experiments for the first time. From the results of experiments on cells with different adherent shapes and different pairs of normal cells and cancer cells, we found additional force provided by membrane tension to be the main component of the force applied to the AFM probe, with decreased cell membrane tension being the essential reason for the greater softness of cancer cells than of normal cells. Hence, regulating membrane tension may become an important method for regulating the behavior of cancer cells.

One-step fabrication of functionalized poly(etheretherketone) surfaces with enhanced biocompatibility and osteogenic activity.

Polyetheretherketone (PEEK) has an elastic modulus similar to that of the bone; however, its use as a material for bone repair is limited by bio-inert surface chemistry and poor osteogenesis-inducing capacity. To address this issue, the PEEK surface was activated by ultraviolet radiation-induced grafting of methacrylated hyaluronic acid (MeHA) and titanium dioxide (TiO2) nanofibers via a one-step process. The modified PEEK surface was characterized by X-ray photoelectron and Fourier-transform infrared spectroscopy, and the extent of surface modification was evaluated by measuring static contact angles. Atomic force microscopy revealed that the PEEK surface grafted with electrospun TiO2 had abundant nanofibers and a roughness that was comparable to that of human cortical bone. In vitro experiment, rat bone mesenchymal stem cells showed increased adhesion, proliferation, and osteogenic differentiation capacity on TiO2-modified as compared to unmodified PEEK. Thus, PEEK that is surface-modified with electrospun TiO2 and MeHA has enhanced biocompatibility and can be an effective material for use in orthopedic implants and medical devices.

Mono-fullerenols modulating cell stiffness by perturbing actin bundling.

Understanding what modulates the cell stiffness is important given its potential application as a diagnostic and medical target. Here, we investigated why and how mono-fullerenols affect the cell stiffness. We confirmed the fullerenol-modulation of cell stiffness using atomic force microscopy (AFM) with sphere tips and ascertained that the particles reduce the cell polarity. The structures of b-actin and f-actin were evaluated by inverted fluorescence microscopy, synchrotron radiation small angle X-ray scattering (SAXS), transmission electron microscopy (TEM) and AFM. Statistical and quantitative analyses of the SAXS data of fullerenol-treated b-actin and f-actin reveal a transformation from large-size to small-size b-actin and simultaneously to f-actin. The slight increase in f-actin diameter in the treated group suggests that fullerenols attach to the actin surface. We verified the attachment using AFM and high-resolution probes. Collectively, our results suggest that fullerenols hamper the bundling of f-actin to form b-actin by adhering to the surface of f-actin, weakening the bundle-based cell stiffness.