Average Rheological Quantities of Cells in Monolayers.

The method of cell monolayer rheology enables quantifying average rheological properties of cell in a single experimental run of few millions cells together in a single layer. Here we describe step-by-step procedure as to how to employ a modified commercial rotational rheometer to run rheological measurement and detect average viscoelastic properties of cells while maintaining the necessary precision level at the same time.

Shear rheology of methyl cellulose based solutions for cell mechanical measurements at high shear rates.

Methyl cellulose (MC) is a widely used material in various microfluidic applications in biology. Due to its biocompatibility, it has become a popular crowding agent for microfluidic cell deformability measurements, which usually operate at high shear rates (>10 000 s-1). However, a full rheological characterization of methyl cellulose solutions under these conditions has not yet been reported. With this study, we provide a full shear-rheological description for solutions of up to 1% MC dissolved in phosphate-buffered saline (PBS) that are commonly used in real-time deformability cytometry (RT-DC). We characterized three different MC-PBS solutions used for cell mechanical measurements in RT-DC with three different shear rheometer setups to cover a range of shear rates from 0.1-150 000 s-1. We report viscosities and normal stress differences in this regime. Viscosity functions can be well described using a Carreau-Yasuda model. Furthermore, we present the temperature dependency of shear viscosity and first normal stress difference of these solutions. Our results show that methyl cellulose solutions behave like power-law liquids in viscosity and exhibit first normal stress difference at shear rates between 5000-150 000 s-1. We construct a general viscosity equation for each MC solution at a certain shear rate and temperature. Furthermore, we investigated how MC concentration influences the rheology of the solutions and found the entanglement concentration at around 0.64 w/w%. Our results help to better understand the viscoelastic behavior of MC solutions, which can now be considered when modelling stresses in microfluidic channels.

Measuring the linear viscoelastic regime of MCF-7 cells with a monolayer rheometer in the presence of microtubule-active anti-cancer drugs at high concentrations.

The rheological properties of cells have vital functional implications. Depending, for instance, on the life cycle, cells show large cell-to-cell variations making it cumbersome to quantify average viscoelastic properties of cells by single-cell techniques. Microfluidic devices, typically working in the nonlinear viscoelastic range, allow fast analysis of single-cell deformation. Averaging over a large number of cells can also be achieved by studying them in a monolayer between rheometer discs. This technique allows applying well-established rheological standard procedures to cell rheology. It offers further advantages like studying cells in the linear viscoelastic range while quantifying cell vitality. Here, we study the applicability of the technique to rather adverse conditions, like for microtubule-active anti-cancer drugs and for a cell line with large size variation. We found a strong impact of the gap width and of normal forces on the moduli and obtained high vitality levels during the rheological study. To enable studying the impact of microtubule-active drugs on vital cells at concentrations several orders of magnitude beyond the half maximal effective concentration for cytotoxicity, we arrested the cell cycle with hydroxyurea. Irrespective of the high concentrations, we observed no clear impact of the microtubule-active drugs.

Narrow-Gap Rheometry: A Novel Method for Measuring Cell Mechanics.

The viscoelastic properties of a cell cytoskeleton contain abundant information about the state of a cell. Cells show a response to a specific environment or an administered drug through changes in their viscoelastic properties. Studies of single cells have shown that chemical agents that interact with the cytoskeleton can alter mechanical cell properties and suppress mitosis. This envisions using rheological measurements as a non-specific tool for drug development, the pharmacological screening of new drug agents, and to optimize dosage. Although there exists a number of sophisticated methods for studying mechanical properties of single cells, studying concentration dependencies is difficult and cumbersome with these methods: large cell-to-cell variations demand high repetition rates to obtain statistically significant data. Furthermore, method-induced changes in the cell mechanics cannot be excluded when working in a nonlinear viscoelastic range. To address these issues, we not only compared narrow-gap rheometry with commonly used single cell techniques, such as atomic force microscopy and microfluidic-based approaches, but we also compared existing cell monolayer studies used to estimate cell mechanical properties. This review provides insight for whether and how narrow-gap rheometer could be used as an efficient drug screening tool, which could further improve our current understanding of the mechanical issues present in the treatment of human diseases.

Physiological and Clinical Aspects of Bioactive Peptides from Marine Animals.

Biological molecules in nutraceuticals and functional foods have proven physiological properties to treat human chronic diseases. These molecules contribute to applications in the food and pharmaceutical industries by preventing food spoilage and cellular injury. Technological advancement in the screening and characterization of bioactive peptides has enabled scientists to understand the associated molecules. Consistent collaboration among nutritionists, pharmacists, food scientists, and bioengineers to find new bioactive compounds with higher therapeutic potential against nutrition-related diseases highlights the potential of the bioactive peptides for food and pharmaceutic industries. Among the popular dietary supplements, marine animals have always been considered imperative due to their rich nutritional values and byproduct use in the food and pharmaceutical industries. The bioactive peptides isolated from marine animals are well-known for their higher bioactivities against human diseases. The physiological properties of fish-based hydrolyzed proteins and peptides have been claimed through in vitro, in vivo, and clinical trials. However, systematic study on the physiological and clinical significance of these bioactive peptides is scarce. In this review, we not only discuss the physiological and clinical significance of antioxidant and anticancer peptides derived from marine animals, but we also compare their biological activities through existing in vitro and in vivo studies.

Visually Based Characterization of the Incipient Particle Motion in Regular Substrates: From Laminar to Turbulent Conditions.

Two different experimental methods for determining the threshold of particle motion as a function of geometrical properties of the bed from laminar to turbulent flow conditions are presented. For that purpose, the incipient motion of a single bead is studied on regular substrates that consist of a monolayer of fixed spheres of uniform size that are regularly arranged in triangular and quadratic symmetries. The threshold is characterized by the critical Shields number. The criterion for the onset of motion is defined as the displacement from the original equilibrium position to the neighboring one. The displacement and the mode of motion are identified with an imaging system. The laminar flow is induced using a rotational rheometer with a parallel disk configuration. The shear Reynolds number remains below 1. The turbulent flow is induced in a low-speed wind tunnel with open jet test section. The air velocity is regulated with a frequency converter on the blower fan. The velocity profile is measured with a hot wire probe connected to a hot film anemometer. The shear Reynolds number ranges between 40 and 150. The logarithmic velocity law and the modified wall law presented by Rotta are used to infer the shear velocity from the experimental data. The latter is of special interest when the mobile bead is partially exposed to the turbulent flow in the so-called hydraulically transitional flow regime. The shear stress is estimated at onset of motion. Some illustrative results showing the strong impact of the angle of repose, and the exposure of the bead to shear flow are represented in both regimes.

Measuring the adhesion limit of fibronectin for fibroblasts with a narrow-gap rotational rheometer.

We study the adhesion limit of 3T6 fibroblasts, cultured in Dulbecco's modified Eagle's medium with 10% fetal bovine serum at 37 °C and 5% CO2, with a narrow-gap rotational rheometer in the parallel-disk configuration. Reducing the uncertainty in gap width to about 1 µm allows studying the cells at narrow gaps, which enables to study the critical shear stress of the cells in low-viscous media. The adhesion limit on fibronectin-coated glass plates is determined as a function of concentration and adhesion time. We found that cells in groups have a tendency to detach at slightly higher shear stresses than single cells. Moreover, 60 min after the settling phase are enough for the cells to adhere to the coated plate at maximum strength. We show that the setup may also be used for cells that are not adhered from suspension, but are grown directly on the substrate.

Average Rheological Quantities of Cells in Monolayers.

Measuring rheological properties of cells in monolayers enables quantifying average cell properties in single experimental runs despite large cell-to-cell variations. Here, we describe how to modify a commercial rotational rheometer to accomplish the necessary precision for a monolayer rheometer and we delineate the steps for setting up experiments detecting average viscoelastic cell properties.

Deformation and rupture of Dunaliella salina at high shear rates without the use of thickeners.

BACKGROUND: High-density cultures require operating below the critical threshold of shear stress, in order to avoid reducing the specific growth rate of the cells. When determining this threshold, direct inspection of the cells in flow provides insight into the conditions of shearing. OBJECTIVE: Aim of this study was using a novel rheo-optical setup for the observation of cells in laminar shear flow and the determination of the critical shear stress required to damage them in their natural environment. METHODS: Dunaliella salina cells were sheared and observed in flow for shear stresses of up to 90 Pa, at ambient temperature, without adding thickeners. The critical shear stress was determined by fitting a hydrodynamics-based criterion to the experimental data on the percentage of deformed cells after shearing. RESULTS: Single cells, clusters and strings of cells were visible in shear flow. The strings formed at maximum shear stresses of 10 Pa or higher. Cells lost motility for maximum shear stresses higher than 15 Pa, and more than 80% of the cells were deformed at maximum shear stresses higher than 60 Pa. The estimated critical shear stress was 18 Pa. CONCLUSIONS: Shear stresses higher than 18 Pa should be avoided when cultivating D. salina.

Polyoxometalate ionic liquids as self-repairing acid-resistant corrosion protection.

Corrosion is a global problem for any metallic structure or material. Herein we show how metals can easily be protected against acid corrosion using hydrophobic polyoxometalate-based ionic liquids (POM-ILs). Copper metal disks were coated with room-temperature POM-ILs composed of transition-metal functionalized Keggin anions [SiW11 O39 TM(H2 O)](n-) (TM=Cu(II) , Fe(III) ) and quaternary alkylammonium cations (Cn H2 n+1 )4 N(+) (n=7-8). The corrosion resistance against acetic acid vapors and simulated "acid rain" was significantly improved compared with commercial ionic liquids or solid polyoxometalate coatings. Mechanical damage to the POM-IL coating is self-repaired in less than one minute with full retention of the acid protection properties. The coating can easily be removed and recovered by rinsing with organic solvents.

Pressure-induced phase transitions in triacylglycerides.

The melting point of triacylglycerides (TAGs) under atmospheric pressure depends on both the fatty acid composition and crystalline structure of the polymorphic state, which are influenced by the temperature treatment history of the TAG. In this contribution, the additional effect of high hydrostatic pressure is described. Samples were placed in a temperature-controlled cell and pressurized up to 450 MPa. The phase transition was investigated either by perpendicular light scattering and transmission or with a polarized-light microscope. The high-pressure polarized-light microscope allows a precise determination of the melting point. The investigated TAGs showed a significant nonlinear increase of the melting point with pressure. Light scattering and transmission were used to observe the phase change in the high-pressure cell. Similar to supercooling in temperature-induced phase transition, we found a dramatic increase of the delay time in our pressure-induced solidification. Even the dependency of this induction time on the control parameter pressure was similar to that in temperature-driven crystallization. We propose that different crystalline structures may be obtained by superpressuring instead of supercooling.

Barchan dunes in two dimensions: experimental tests for minimal models.

A well-defined two-dimensional single barchan dune under the force of a shearing water flow is investigated experimentally. From an initially prepared triangular heap a rapid relaxation to a steady-state solution is observed with constant mass, shape, and velocity. This attractor exhibits all characteristic features of barchan dunes found in nature, namely a gently inclined windward side, crest, brink, and steep lee face. The relaxation time towards the steady state increases with mass. For small dunes we find significant deviations from a fixed height-length aspect ratio. As predicted by recent theoretical models, the migration velocity scales reciprocal to the length of the dune.