A biomechanical model for the transendothelial migration of cancer cells

We propose a biomechanical model for the extravasation of a tumor cell (TC) through the endothelium of a blood vessel. Based on prior in vitro observations, we assume that the TC extends a protrusion between adjacent endothelial cells (ECs) that adheres to the basement membrane via focal adhesions. As the protrusion grows in size and branches out, the actomyosin contraction along the stress fibers inside the protrusion pulls the relatively rigid nucleus through the endothelial opening. We model the chemo-mechanics of the stress fibers and the focal adhesions by following the kinetics of the active myosin motors and high-affinity integrins, subject to mechanical feedback. This is incorporated into a finite-element simulation of the extravasation process, with the contractile force pulling the nucleus of the tumor cell against elastic resistance of the ECs. To account for the interaction between the TC nucleus and the endothelium, we consider two scenarios: solid-solid contact and lubrication by cytosol. The former gives a lower bound for the required contractile force to realize transmigration, while the latter provides a more realistic representation of the process. Using physiologically reasonable 1 parameters, our model shows that the stress-fiber and focal-adhesion ensemble can produce a contractile force on the order of 70 nN, which is sufficient to deform the ECs and enable transmigration. Furthermore, we use an atomic force microscope to measure the resistant force on a human bladder cancer cell that is pushed through an endothelium cultured in vitro. The magnitude of the required force turns out to be in the range of 70-100 nN, comparable to the model predictions

Paroxysmal Permeability Disorders: Development of a Microfluidic Device to Assess Endothelial Barrier Function

Background: Paroxysmal Permeability Disorders (PPDs) are pathological conditions caused by periodic short lasting increase of endothelial permeability, in the absence of inflammatory, degenerative, ischemic vascular injury. PPDs include primary angioedema, idiopathic systemic capillary leak syndrome and some rare forms of localized retroperitoneal-mediastinal edema.Aim: to validate a microfluidic device to study endothelial permeability in flow conditions.Materials and Methods: we designed a microchannel network (the smallest channel is 30μm square section). Human Umbilical Vein Endothelial Cells (HUVECs) were cultured under constant shear stress in the networks. Endothelial permeability assessment was based on interaction of biotinylated fibronectin used as a matrix for HUVECs and FITC-conjugated avidin. The increase in endothelial permeability was identified as changes in fluorescence intensity detected by confocal fluorescent microscopy.Results: The microchannels were constantly perfused with a steady flow of culture medium, ensuring a physiologically relevant level of shear stress at the wall of ~0.2 Pa. Our preliminary results demonstrated that circulation of culture medium or plasma from healthy volunteers was associated with low fluorescence of fibronectin matrix. When bradykinin diluted in culture medium was perfused, an increase in average fluorescence was detected.Conclusion: Our microvasculature model is suitable to study endothelial functions in physiological flow conditions and in the presence of factors like bradykinin known as mediator of several PPDs. Therefore, it can be a promising tool to better understand the mechanisms underlying disorders of endothelial permeability

The intriguing role of collagen on the rheology of cancer cell spheroids

International audienceSpheroids are multicellular systems with an interesting rheology giving rise to elasto–visco–plastic properties. They are good tumor models, but the role of the extracellular matrix (ECM) is not fully understood. ECM is an important link between cells and may have a significant impact on tissue organization. Here we determine viscoelastic properties of spheroids including different collagen I amounts using AFM and predict new frequency–dependent properties leading to soft glassy rheology behavior. A unified model – similar to single cell behavior – is proposed and discussed, while complementary confocal experiments reveal the microstructure of spheroids, with collagen I fibers serving as a skeleton for cells, thus reinforcing the spheroid viscoelastic behavior

Changes in endothelial cell microrheology in medium modified by cancer cells

During the process of metastasis, cancer cells are detached from the primary tumour and may find their way into the blood system, from where they can migrate in a distant site. Endothelial cells line the vascular wall of all blood and lymphatic vessels and regulate its permeability for substances and migratory cells. Previous studies have focused on the properties of cancer cells (Abidine et al. 2018), as well as their interaction with endothelial cells (Rajan et al. 2016), as they attach to and transmigrate through the endothelium barrier. For many authors, the most widely accepted hypothesis was that transmigration is a characteristic of cancer cells, overlooking the possibility of endothelial cells actively participating in this process (Mierke et al. 2012). In this paper, endothelial monolayers were studied with a previously developed AFM microrheology technique (Abidine et al. 2018) when cultured in cancer cell conditioned medium, at different dilutions. This medium is enriched with molecules secreted by the cancer cells, which may play a role during the cancer cells’ migration and alter the endothelial cells’ behaviour (Ritchie et al. 2021). We show the first results of the viscoelastic properties of endothelial cells under these experimental conditions

The effect of shear stress reduction on endothelial cells: a microfluidic study of the actin cytoskeleton

International audienceReduced blood flow, as occurring in ischemia or resulting from exposure to microgravity such as encountered in space flights, induces a decrease in the level of shear stress sensed by the endothelial cells forming the inner part of blood vessels. In the present study, we use a microvasculature-on-a-chip device in order to investigate in vitro the effect of such a reduction in shear stress on shear-adapted endothelial cells. We find that, within one hour of exposition to reduced wall shear stress, human umbilical vein endothelial cells undergo a reorganization of their actin skeleton, with a decrease in the number of stress fibers and actin being recruited into the cells' peripheral band, indicating a fairly fast change in cells' phenotype due to altered flow

Paroxysmal Permeability Disorders: Development of a Microfluidic Device to Assess Endothelial Barrier Function

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Blood flow and microgravity

International audienceThe absence of gravity during space flight can alter cardio-vascular functions partially due to reduced physical activity. This affects the overall hemodynamics, and in particular the level of shear stresses to which blood vessels are submitted. Long-term exposure to space environment is thus susceptible to induce vascular remodeling through a mechanotransduction cascade that couples vessel shape and function with the mechanical cues exerted by the circulating cells on the vessel walls. Central to such processes, the glycocalyx – i.e. the micron-thick layer of biomacromolecules that lines the lumen of blood vessels and is directly exposed to blood flow – is a major actor in the regulation of biochemical and mechanical interactions. We discuss in this article several experiments performed under microgravity, such as the determination of lift force and collective motion in blood flow, and some preliminary results obtained in artificial microfluidic circuits functionalized with endothelium that offer interesting perspectives for the study of the interactions between blood and endothelium in healthy condition as well as by mimicking the degradation of glycocalyx caused by long space missions. A direct comparison between experiments and simulations is discussed

Rheology of a confined vesicle suspension

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