Vascular surveillance by haptotactic blood platelets in inflammation and infection

Breakdown of vascular barriers is a major complication of inflammatory diseases. Anucleate platelets form blood-clots during thrombosis, but also play a crucial role in inflammation. While spatio-temporal dynamics of clot formation are well characterized, the cell-biological mechanisms of platelet recruitment to inflammatory micro-environments remain incompletely understood. Here we identify Arp2/3-dependent lamellipodia formation as a prominent morphological feature of immune-responsive platelets. Platelets use lamellipodia to scan for fibrin(ogen) deposited on the inflamed vasculature and to directionally spread, to polarize and to govern haptotactic migration along gradients of the adhesive ligand. Platelet-specific abrogation of Arp2/3 interferes with haptotactic repositioning of platelets to microlesions, thus impairing vascular sealing and provoking inflammatory microbleeding. During infection, haptotaxis promotes capture of bacteria and prevents hematogenic dissemination, rendering platelets gate-keepers of the inflamed microvasculature. Consequently, these findings identify haptotaxis as a key effector function of immune-responsive platelets

Alignment of cell division axes in directed epithelial cell migration

Cell division is an essential dynamic event in tissue remodeling during wound healing, cancer and embryogenesis. In collective migration, tensile stresses affect cell shape and polarity, hence, the orientation of the cell division axis is expected to depend on cellular flow patterns. Here, we study the degree of orientation of cell division axes in migrating and resting epithelial cell sheets. We use microstructured channels to create a defined scenario of directed cell invasion and compare this situation to resting but proliferating cell monolayers. In experiments, we find a strong alignment of the axis due to directed flow while resting sheets show very weak global order, but local flow gradients still correlate strongly with the cell division axis. We compare experimental results with a previously published mesoscopic particle based simulation model. Most of the observed effects are reproduced by the simulations

Ion-mediated changes of supported lipid bilayers and their coupling to the substrate. A case of bilayer slip?

Ion-mediated (Ca(2+)) changes in viscoelastic, structural and optical properties of negatively charged solid supported lipid bilayers (SLBs) on SiO(2) surfaces were studied by means of quartz crystal microbalance with dissipation (QCM-D) monitoring and optical reflectometry. Despite the sensitivity of QCM-D to viscoeleastic/structural variations, it has not often been used to probe such changes for SLBs. SLBs were prepared from binary phospholipid mixtures of 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC, neutral) and 1-palmitoyl-2-oleoyl-sn-glycero-3-phospho-(1'-rac-glycerol) (POPG, negatively charged) on SiO(2) sensor surfaces in a Ca(2+)-containing buffer. Interestingly, for bilayers containing POPG fractions above 35%, large QCM-D dissipation shifts occurred, when Ca(2+) was removed from buffer in contact with the SLB (while maintaining 100 mM NaCl). The accompanying frequency changes were small. These Ca(2+) mediated QCM-D responses are reversible, and a signal for considerable changes in the viscoelastic and structural properties of the SLB. Variation of Ca(2+)-concentration revealed a threshold concentration of around 0.4 mM for the changes in the SLB to occur. Below this value, at >35% POPG concentration in the SLB, the SLB appears to become more weakly attached to the SiO(2) substrate, which is partly attributed to a weakening of the POPG-substrate interaction in the absence of Ca(2+). A consequence of this is an oscillation-amplitude dependent dissipation, which we attribute to slip of the bilayer at higher oscillation amplitudes. Complementary experiments using a combined QCM-D/reflectometry instrument showed that the Ca(2+)-induced changes in the viscoelastic/structural properties of the SLB are accompanied by changes in the optical properties. We discuss different scenarios to explain the observed reversible effect of Ca(2+)-ions on the dissipative and optical properties of the mixed SLBs. Based on our results we propose the observed phenomenon to be a combination of geometric changes, internal structural changes, changes in the interfacial water layer, and a slip mechanism, i.e. friction between the SLB and the substrate

Inhibitory effects of trans-resveratrol analogs molecules on the proliferation and the cell cycle progression of human colon tumoral cells.

International audienceResveratrol may function as a cancer chemopreventive agent. However, few data are available on the antitumoral activities of its dimer, epsilon-viniferin, also present in human diet. So, the effects of resveratrol, epsilon-viniferin, of their acetylated forms (resveratrol triacetate, epsilon-viniferin pentaacetate) and of vineatrol (a wine grape extract) were compared on human adenocarcinoma colon cells. Resveratrol and resveratrol triacetate inhibit cell proliferation and arrest cell cycle. epsilon-Viniferin and epsilon-viniferin pentaacetate slightly reduce cell proliferation. Vineatrol inhibits cell proliferation and favors an accumulation in the S phase of the cell cycle. Consequently, resveratrol triacetate and vineatrol could constitute new putative anticancer agents on colon carcinoma

Phenomenological approaches to collective behavior in epithelial cell migration

AbstractCollective cell migration in epithelial tissues resembles fluid-like behavior in time-lapse recordings. In the last years, hydrodynamic velocity fields in living matter have been studied intensely. The emergent properties were remarkably similar to phenomena known from active soft matter systems. Here, we review migration experiments of large cellular ensembles as well as of mesoscopic cohorts in micro-structured environments. Concepts such as diffusion, velocity correlations, swirl strength and polarization are metrics to quantify the cellular dynamics both in experiments as well as in computational simulations. We discuss challenges relating collective migration to single cell and oligocellular behavior as well as linking the phenotypic parameters to the underlying cytoskeleton dynamics and signaling networks. This article is part of a Special Issue entitled: Mechanobiology

Dynamic Changes of Acoustic Load and Complex Impedance as Reporters for the Cytotoxicity of Small Molecule Inhibitors

Cellular motility is the major driving force of numerous biological phenomena including wound healing, immune response, embryogenesis, cancer formation, and metastasis. We studied the response of epithelial FaDu monolayers cultured on gold electrodes of an acoustic resonator (quartz crystal microbalance, QCM) and impedance sensor (electric cell-substrate impedance sensing, ECIS) to externally applied chemical stimuli interfering with cytoskeleton organization. Epithelial cell motility of confluent monolayers is characterized by subtle cell shape changes and variations in the cell-substrate as well as cell-cell distance without net directionality of individual cells. The impact of small molecules such as cytochalasin D, phalloidin, and blebbistatin as well as paclitaxel, nocodazol, and colchicin on actin and microtubules organization was quantified by conventional sensors\u27 readouts and by comparing the noise pattern of the signals which is attributed to cellular dynamics. The responsiveness of noninvasive and label-free techniques relying on cellular dynamics is compared to classical viability assays and changes of the overall impedance of ultrasmall electrodes or acoustic loads of a thickness shear mode resonator. Depending on the agent used, a distinct sensor response was found, which can be used as a fingerprint of the cellular response. Cytoskeletal rearrangements and nuclear integrity were corroborated by fluorescence microscopy and correlated to the readouts of QCM and ECIS

Dynamic Changes of Acoustic Load and Complex Impedance as Reporters for the Cytotoxicity of Small Molecule Inhibitors

Cellular motility is the major driving force of numerous biological phenomena including wound healing, immune response, embryogenesis, cancer formation, and metastasis. We studied the response of epithelial FaDu monolayers cultured on gold electrodes of an acoustic resonator (quartz crystal microbalance, QCM) and impedance sensor (electric cell-substrate impedance sensing, ECIS) to externally applied chemical stimuli interfering with cytoskeleton organization. Epithelial cell motility of confluent monolayers is characterized by subtle cell shape changes and variations in the cell-substrate as well as cell-cell distance without net directionality of individual cells. The impact of small molecules such as cytochalasin D, phalloidin, and blebbistatin as well as paclitaxel, nocodazol, and colchicin on actin and microtubules organization was quantified by conventional sensors\u27 readouts and by comparing the noise pattern of the signals which is attributed to cellular dynamics. The responsiveness of noninvasive and label-free techniques relying on cellular dynamics is compared to classical viability assays and changes of the overall impedance of ultrasmall electrodes or acoustic loads of a thickness shear mode resonator. Depending on the agent used, a distinct sensor response was found, which can be used as a fingerprint of the cellular response. Cytoskeletal rearrangements and nuclear integrity were corroborated by fluorescence microscopy and correlated to the readouts of QCM and ECIS

Extrusion texturization of cricket flour and soy protein isolate: Influence of insect content, extrusion temperature, and moisture-level variation on textural properties

Due to the increasing global population and unsustainable meat production, the future supply of animal-derived protein is predicted to be insufficient. Currently, edible insects are considered as a potential and “novel” source of protein in the development of palatable meat analogues. This research used high moisture extrusion cooking (HMEC), at a screw speed of 150 rpm, to produce meat analogues using full- or low-fat cricket flours (CF) and soy protein isolate (SPI). Effects of water flow rate (WFR), cooking temperature (9 and 10 ml/min; 120, 140, and 160°C, respectively), and CF inclusions levels of 0, 15, 30, and 45% were analyzed. Cooking temperature and CF inclusion had a significant effect (p < .05) on both tensile stress in parallel and perpendicular directions, while WFR had no significant effect (p = .3357 and 0.7700), respectively. The tensile stress increased with temperature but decreased with CF inclusion at both WFRs. Comparatively, the tensile stress was stronger at WFR of 9 ml/min than at 10 ml/min; however, the tensile stress in parallel was mostly greater than tensile stress in perpendicular directions. Fibrous meat analogues with high anisotropic indices (AIs) of up to 2.80 were obtained, particularly at WFR of 10 ml/min and at inclusions of 30% low-fat CF. By controlling HMEC conditions, full-/low-fat cricket flours at 15% and 30% inclusions can offer an opportunity to partially substitute SPI in manufacturing of fibrous meat analogues