Viscoelastic properties of suspended cells measured with shear flow deformation cytometry

Numerous cell functions are accompanied by phenotypic changes in viscoelastic properties, and measuring them can help elucidate higher-level cellular functions in health and disease. We present a high-throughput, simple and low-cost microfluidic method for quantitatively measuring the elastic (storage) and viscous (loss) modulus of individual cells. Cells are suspended in a high-viscosity fluid and are pumped with high pressure through a 5.8 cm long and 200 μm wide microfluidic channel. The fluid shear stress induces large, near ellipsoidal cell deformations. In addition, the flow profile in the channel causes the cells to rotate in a tank-treading manner. From the cell deformation and tank treading frequency, we extract the frequency-dependent viscoelastic cell properties based on a theoretical framework developed by R. Roscoe that describes the deformation of a viscoelastic sphere in a viscous fluid under steady laminar flow. We confirm the accuracy of the method using atomic force microscopy-calibrated polyacrylamide beads and cells. Our measurements demonstrate that suspended cells exhibit power-law, soft glassy rheological behavior that is cell cycle-dependent and mediated by the physical interplay between the actin filament and intermediate filament networks.

Atlas-Based Evaluation of Hemodynamic in Ascending Thoracic Aortic Aneurysms

Atlas-based analyses of patients with cardiovascular diseases have recently been explored to understand the mechanistic link between shape and pathophysiology. The construction of probabilistic atlases is based on statistical shape modeling (SSM) to assess key anatomic features for a given patient population. Such an approach is relevant to study the complex nature of the ascending thoracic aortic aneurysm (ATAA) as characterized by different patterns of aortic shapes and valve phenotypes. This study was carried out to develop an SSM of the dilated aorta with both bicuspid aortic valve (BAV) and tricuspid aortic valve (TAV), and then assess the computational hemodynamic of virtual models obtained by the deformation of the mean template for specific shape boundaries (i.e., ±1.5 standard deviation, σ). Simulations demonstrated remarkable changes in the velocity streamlines, blood pressure, and fluid shear stress with the principal shape modes such as the aortic size (Mode 1), vessel tortuosity (Mode 2), and aortic valve morphologies (Mode 3). The atlas-based disease assessment can represent a powerful tool to reveal important insights on ATAA-derived hemodynamic, especially for aneurysms which are considered to have borderline anatomies, and thus challenging decision-making. The utilization of SSMs for creating probabilistic patient cohorts can facilitate the understanding of the heterogenous nature of the dilated ascending aorta.

Coupled CFD-DEM modelling to predict how EPS affects bacterial biofilm deformation, recovery and detachment under flow conditions

The deformation and detachment of bacterial biofilm are related to the structural and mechanical properties of the biofilm itself. Extracellular polymeric substances (EPS) play an important role on keeping the mechanical stability of biofilms. The understanding of biofilm mechanics and detachment can help to reveal biofilm survival mechanisms under fluid shear and provide insight about what flows might be needed to remove biofilm in a cleaning cycle or for a ship to remove biofilms. However, how the EPS may affect biofilm mechanics and its deformation in flow conditions remains elusive. To address this, a coupled computational fluid dynamic – discrete element method (CFD-DEM) model was developed. The mechanisms of biofilm detachment, such as erosion and sloughing have been revealed by imposing hydrodynamic fluid flow at different velocities and loading rates. The model, which also allows adjustment of the proportion of different functional group of microorganisms in the biofilm, enables the study of the contribution of EPS towards biofilm resistance to fluid shear stress. Furthermore, the stress-strain curves during biofilm deformation have been captured by loading and unloading fluid shear stress to study the viscoelastic properties of the biofilm.

MEKK3–TGFβ crosstalk regulates inward arterial remodeling

Arterial remodeling is an important adaptive mechanism that maintains normal fluid shear stress in a variety of physiologic and pathologic conditions. Inward remodeling, a process that leads to reduction in arterial diameter, plays a critical role in progression of such common diseases as hypertension and atherosclerosis. Yet, despite its pathogenic importance, molecular mechanisms controlling inward remodeling remain undefined. Mitogen-activated protein kinases (MAPKs) perform a number of functions ranging from control of proliferation to migration and cell-fate transitions. While the MAPK ERK1/2 signaling pathway has been extensively examined in the endothelium, less is known about the role of the MEKK3/ERK5 pathway in vascular remodeling. To better define the role played by this signaling cascade, we studied the effect of endothelial-specific deletion of its key upstream MAP3K, MEKK3, in adult mice. The gene’s deletion resulted in a gradual inward remodeling of both pulmonary and systematic arteries, leading to spontaneous hypertension in both vascular circuits and accelerated progression of atherosclerosis in hyperlipidemic mice. Molecular analysis revealed activation of TGFβ-signaling both in vitro and in vivo. Endothelial-specific TGFβR1 knockout prevented inward arterial remodeling in MEKK3 endothelial knockout mice. These data point to the unexpected participation of endothelial MEKK3 in regulation of TGFβR1-Smad2/3 signaling and inward arterial remodeling in artery diseases.

MULTI-SCALE MECHANICAL BEHAVIOR ANALYSIS ON FLUID–SOLID COUPLING FOR OSTEONS IN VARIOUS GRAVITATIONAL FIELDS

Background: Compact bone mainly consists of cylindrical osteon structures. In microgravity, the change in the mechanical microenvironment of osteocytes might be the root cause of astronauts’ bone loss during space flights. Methods: A multi-scale three-dimensional (3D) fluid–solid coupling finite element model of osteons with a two-stage pore structure was developed using COMSOL software based on the natural structure of osteocytes. Gradients in gravitational fields of [Formula: see text]1, 0, 1, 2.5, and 3.7[Formula: see text]g were used to investigate the changes in the mechanical microenvironment on osteocyte structure. The difference in arteriole pulsating pressure and static compression stress caused by each gravity gradient was investigated. Results: The mechanical response of osteocytes increased with the value of g, compared with the Earth’s gravitational field. For instance, the fluid pressure of osteocytes and the von Mises stress of bone matrix near lacunae decreased by 31.3% and 99.9%, respectively, in microgravity. Under static loading, only about 16.7% of osteocytes in microgravity and 58.3% of osteocytes in the Earth’s gravitational field could reach the fluid shear stress threshold of biological reactions in cell culture experiments. Compared with the Earth’s gravitational field, the pressure gradient inside osteocytes severely decreased in microgravity. Conclusion: The mechanical microenvironment of osteocytes in microgravity might cause significant changes in the mechanical microenvironment of osteocytes, which may lead to disuse osteoporosis in astronauts.

Fluid-Solid Coupling Simulation of Wall Fluid Shear Stress on Cells under Gradient Fluid Flow

Fluid shear stress (FSS) plays a crucial role for cell migration within bone cavities filled with interstitial fluid. Whether the local wall FSS distribution on cell surface depends on the global gradient FSS of flow field should be clarified to explain our previous experimental observation. In this study, finite element models of discretely distributed or hexagonal closely packed cells adherent on the bottom plate in a modified plate flow chamber with different global FSS gradient were constructed. Fluid-solid coupling simulation of wall fluid shear stress on cells was performed, and two types of data analysis methods were used. The results showed that the profile of local FSS distribution on cell surface coincides with the angle of cell migration determined in the previous study, suggesting that RAW264.7 osteoclast precursors may sense the global FSS gradient and migrate toward the low-FSS region under a high gradient. For hexagonal closely packed cells, this profile on the surface of central cells decreased along with the increase of cell spacing, which may be caused by the higher local FSS difference along the direction of FSS gradient in the regions close to the bottom plate. This study may explain the phenomenon of the targeted migration of osteoclast precursors under gradient FSS field and further provide insights into the mechanism of mechanical stimulation-induced bone remodeling.

Therapeutic Effect of Platinum Nanoparticles on Atherosclerosis Research Model Based on Microfluidics and Determination of Injury

The atherosclerosis (AS) microenvironment plays an important role in pathogenicity, including blood flow and blood pressure, high cholesterol, high blood sugar, angiotensin II, tumor necrosis factor, and the like. The AS microfluidic model was established, and the fluid shear stress and cyclic stretching were set to 5.07 Pa and 1.17 Hz to simulate normal blood flow, respectively. The effects of different biochemical environments on endothelial cells (ECs) and cardiomyocytes were analyzed. The results confirmed that different biochemical environments had significant damage to ECs and cardiomyocytes. Subsequently, the further effect of ECs on cardiomyocytes in AS microenvironment was studied, and the results proved that ECs caused further damage to cardiomyocytes under AS biochemical factors. We used Pt nanoparticles (Pt NPs) to study the anti-AS efficiency. The results showed that the addition of Pt NPs played a particular role in the AS treatment of ECs in the AS microenvironment, and the protection for myocardial cells was achieved.

The Response of Corneal Endothelial Cells to Shear Stress in an In Vitro Flow Model

Purpose. Corneal endothelial cells are usually exposed to shear stress caused by the aqueous humour, which is similar to the exposure of vascular endothelial cells to shear stress caused by blood flow. However, the effect of fluid shear stress on corneal endothelial cells is still poorly understood. The purpose of this study was to explore whether the shear stress that results from the aqueous humour influences corneal endothelial cells. Methods. An in vitro model was established to generate fluid flow on cells, and the effect of fluid flow on corneal endothelial cells after exposure to two levels of shear stress for different durations was investigated. The mRNA and protein expression of corneal endothelium-related markers in rabbit corneal endothelial cells was evaluated by real-time PCR and western blotting. Results. The expression of the corneal endothelium-related markers ZO-1, N-cadherin, and Na+-K+-ATPase in rabbit corneal endothelial cells (RCECs) was upregulated at both the mRNA and protein levels after exposure to shear stress. Conclusion. This study demonstrates that RCECs respond favourably to fluid shear stress, which may contribute to the maintenance of corneal endothelial cell function. Furthermore, this study also provides a theoretical foundation for further investigating the response of human corneal endothelial cells to the shear stress caused by the aqueous humour.

PGC1α Regulates the Endothelial Response to Fluid Shear Stress via Telomerase Reverse Transcriptase Control of Heme Oxygenase-1

Objective: Fluid shear stress (FSS) is known to mediate multiple phenotypic changes in the endothelium. Laminar FSS (undisturbed flow) is known to promote endothelial alignment to flow, which is key to stabilizing the endothelium and rendering it resistant to atherosclerosis and thrombosis. The molecular pathways responsible for endothelial responses to FSS are only partially understood. In this study, we determine the role of PGC1α (peroxisome proliferator gamma coactivator-1α)-TERT (telomerase reverse transcriptase)-HMOX1 (heme oxygenase-1) during shear stress in vitro and in vivo. Approach and Results: Here, we have identified PGC1α as a flow-responsive gene required for endothelial flow alignment in vitro and in vivo. Compared with oscillatory FSS (disturbed flow) or static conditions, laminar FSS (undisturbed flow) showed increased PGC1α expression and its transcriptional coactivation. PGC1α was required for laminar FSS-induced expression of TERT in vitro and in vivo via its association with ERRα(estrogen-related receptor alpha) and KLF (Kruppel-like factor)-4 on the TERT promoter. We found that TERT inhibition attenuated endothelial flow alignment, elongation, and nuclear polarization in response to laminar FSS in vitro and in vivo. Among the flow-responsive genes sensitive to TERT status, HMOX1 was required for endothelial alignment to laminar FSS. Conclusions: These data suggest an important role for a PGC1α-TERT-HMOX1 axis in the endothelial stabilization response to laminar FSS.