Effect Of Matrix Elasticity Research Articles

Gold Nanowires/Fibrin Nanostructure as Microfluidics Platforms for Enhancing Stem Cell Differentiation: Bio-AFM Study.

Organ-on-a-chip technology has gained great interest in recent years given its ability to control the spatio-temporal microenvironments of cells and tissues precisely. While physical parameters of the respective niche such as microchannel network sizes, geometric features, flow rates, and shear forces, as well as oxygen tension and concentration gradients, have been optimized for stem cell cultures, little has been done to improve cell-matrix interactions in microphysiological systems. Specifically, detailed research on the effect of matrix elasticity and extracellular matrix (ECM) nanotopography on stem cell differentiation are still in its infancy, an aspect that is known to alter a stem cell’s fate. Although a wide range of hydrogels such as gelatin, collagen, fibrin, and others are available for stem cell chip cultivations, only a limited number of elasticities are generally employed. Matrix elasticity and the corresponding nanotopography are key factors that guide stem cell differentiation. Given this, we investigated the addition of gold nanowires into hydrogels to create a tunable biointerface that could be readily integrated into any organ-on-a-chip and cell chip system. In the presented work, we investigated the matrix elasticity (Young’s modulus, stiffness, adhesive force, and roughness) and nanotopography of gold nanowire loaded onto fibrin hydrogels using the bio-AFM (atomic force microscopy) method. Additionally, we investigated the capacity of human amniotic mesenchymal stem cells (hAMSCs) to differentiate into osteo- and chondrogenic lineages. Our results demonstrated that nanogold structured-hydrogels promoted differentiation of hAMSCs as shown by a significant increase in Collagen I and II production. Additionally, there was enhanced calcium mineralization activity and proteoglycans formation after a cultivation period of two weeks within microfluidic devices.

Impact of Hydrogel Elasticity and Adherence on Osteosarcoma Cells and Osteoblasts

Biochemical and physical properties of extracellular matrix (ECM) control cell behaviors, but how they affect osteosarcoma cells that do not require attachment and their normal counterparts (osteoblasts) that are anchorage-dependent has not been reported yet. In this study, the effects of matrix elasticity and adherence on osteosarcoma MG63 cells are investigated using four types of scaffolds (collagen type I, matrigel, alginate, and agarose) with varied adhesion ligands and rigidity, as compared with osteoblast hFOB1.19 cells. MG63 cells on 2D films are sensitive to ECM adherence, similar to the situation of hFOB1.19 cultured in both 2D and 3D. However, osteosarcoma cells in 3D hydrogels are sensitive to ECM elasticity rather than adherence, with tumor proliferation and malignancy varied with matrix rigidity. The results indicate that osteosarcomas cells might adopt unnatural characteristics on flat surfaces. But in 3D culture, they recover their normal state independent of adherence, as regulated mainly by ECM elasticity via the integrin-mediated focal adhesion pathway, which is further confirmed by in vivo studies. In contrast, osteoblasts and 2D cultured osteosarcoma cells are predominantly influenced by ECM bioactivity regulated by integrin-mediated adherens junction pathway. This study might provide new insights into rational design of scaffolds for tumor/tissue engineering.

Confined tube flow of low viscosity emulsions: Effect of matrix elasticity

This study is addressed to the effect of the elasticity of the continuous phase on confined flow of low viscosity emulsions, a topic of interest in several applications. A Newtonian silicone oil was used as the dispersed phase, while three different water-based fluids, a Newtonian and two Boger fluids, were used as the continuous phase. All the fluids have the same viscosity (ca. 0.05 Pa s). Emulsions were prepared by feeding the two phases to a T-junction followed by a membrane. When the continuous phase was a Boger fluid, the average drop size of the so obtained emulsions was higher (as compared to the Newtonian continuous phase) and a bimodal distribution was observed. Such lower mixing efficiency for a Boger continuous phase is in line with the result for an isolated droplet that breakup is hindered by matrix elasticity. High-speed microscopy visualization of the emulsions flowing through a microcapillary allowed one to determine the velocity profiles and the droplet spatial distribution as a function of the shear rate. In all the experiments, the formation of a droplet-rich core and of a droplet-devoid wall region Δ was observed, with a significant enhancement when the continuous phase was a Boger fluid (where values of Δ as high as 37% of capillary radius were found). So, confinement here refers to the fact that, although the droplets are much smaller than capillary radius, the droplet-free layer takes up a significant fraction of the capillary radius. The observed nonhomogeneous droplet spatial distribution was not due to a transient entry effect, as shown by imaging the emulsions flow at different sections from the capillary inlet, but can be explained by a balance between collision-induced drop diffusion and wall lift force. The former acts to homogenize droplet spatial distribution, while the latter tends to push droplets away from the wall and is enhanced by matrix elasticity. These effects lead to the accumulation of larger droplets around the center of the channel, while smaller droplets are pushed toward the periphery. The velocity profiles for the Boger continuous phase were flat in the droplet-rich core, thus suggesting that the confined emulsions behave as a Bingham fluid. The corresponding yield stress was determined by data fitting of the velocity profiles and rather large values were so obtained. This result is consistent with the high values, close to the maximum packing for spherical particles in binary dispersions, of the droplet volume fraction in the core, which were estimated by measuring the thickness of the droplet-devoid annular region as a function of the applied flow rate and making a volume balance. Overall, the results of this work bear strong analogies with the confined flow of other concentrated suspensions of deformable particles, such as red blood cells in microcirculation, where the formation of a cell-free layer at the wall, which is referred to as margination, has important pathophysiological implications. These findings suggest that low viscosity emulsions with an elastic matrix under confined flow can be used as a tunable model system of biomedical relevance.

Consequences of the effect of matrix elasticity on the rotation of droplet pairs for collision efficiency

The theory of flow-induced coalescence in viscoelastic matrix considering the effect of the matrix elasticity on the approach and rotation of coalescing droplets is derived. The study compares collision efficiencies, i.e. probabilities that the droplet collision will be followed by their fusion, Pc, calculated by means of equations for the droplet rotation in viscoelastic and Newtonian matrixes. It has been found that Pc slightly increases with relaxation time of the matrix for shear flow-induced coalescence, but strongly decreases for extensional flow-induced coalescence.

Effects of matrix elasticity and cell density on human mesenchymal stem cells differentiation

Human mesenchymal stem cells (hMSCs) can differentiate into various cell types, including osteogenic and chondrogenic cells. The matrix elasticity and cell seeding density are important factors in hMSCs differentiation. We cultured hMSCs at different seeding densities on polyacrylamide hydrogels with different stiffness corresponding to Young's moduli of 1.6 ± 0.3 and 40 ± 3.6 kPa. The promotion of osteogenic marker expression by hard gel is overridden by a high seeding density. Cell seeding density, however, did not influence the chondrogenic marker expressions induced by soft gel. These findings suggest that interplays between cell-matrix and cell-cell interactions contribute to hMSCs differentiation. The promotion of osteogenic differentiation on hard matrix was shown to be mediated through the Ras pathway. Inhibition of Ras (RasN17) significantly decreased ERK, Smad1/5/8 and AKT activation, and osteogenic markers expression. However, constitutively active Ras (RasV12) had little effect on osteogenic marker expression, suggesting that the Ras pathways are necessary but not sufficient for osteogenesis. Taken together, our results indicate that matrix elasticity and cell density are important microenvironmental cues driving hMSCs proliferation and differentiation.

Effect of Matrix Elasticity on the Maintenance of the Chondrogenic Phenotype

The aim of this study was to examine the influence of matrix elasticity on the maintenance of the chondrogenic phenotype of chondrocytes cultured in monolayer. We used a two-dimensional culturing system in which polyacrylamide gels with different concentrations of bis-acrylamide were coated with collagen type I. Matrices with a Young's modulus of 4, 10, 40, and 100 kPa were produced, as determined by atomic force microscopy. Porcine chondrocytes were cultivated on these matrices at a low density for 7 days. The proliferation of cells was analyzed by 5-Bromo-2'-deoxy-uridine incorporation. Maintenance of the chondrogenic phenotype was analyzed by measuring collagen type I, type II, and aggrecan gene expression, immunofluorescence staining for collagen type II, and phalloidin staining for actin filaments. Cellular proliferation and actin organization were decreased on matrices of 4 kPa compared with stiffer substrates. The differentiated phenotype of the chondrocytes grown on matrices of 4 kPa was stabilized, indicated by higher collagen type II and aggrecan, and lower collagen type I expression. These findings indicate that chondrocytes sense the elasticity of the matrix and might be used for the design of scaffolds with mechanical properties specifically tailored to support the chondrogenic phenotype in tissue engineering applications.

Towards cellular therapy with a spring in your step!

Surprisingly little is known regarding the effects of the physical microenvironmental niche on haemopoietic stem cells. We have explored the effects of matrix elasticity on stem cell properties using a unique synthetic substrate, tropoelastin, which we used to show that both murine and human primitive haemopoietic cells are more efficiently maintained on an elastically extensible substrate than controls. We have shown that culturing primitive murine and human cord blood haemopoietic cells on tropoelastin in vitro led to increases in cell number, clonogenicity, proliferation and engraftment following transplantation. Atomic force measurements of truncations and crosslinked tropoelastin strengthen the hypothesis that stem cells sense the elasticity of their microenvironment. A consequence of this idea is that alterations to niche elasticity in disease states may contribute to abnormal haemopoiesis. Further, elastic substrates such as tropoelastin may offer a new approach to biomaterial design aimed to achieve optimal ex vivo culture conditions for primitive haemopoietic cells. In order to take advantage of this type of observation arising from the basic research laboratory (and many other opportunities in cellular therapies), a basic introduction in the principles and regulatory aspects of good manufacturing practice of cellular therapeutics will be provided.

Effect of Matrix Elasticity on the Continuous Foaming of Food Models

The aim is to understand the effect of matrix elasticity on continuous foaming using food models based on glucose syrup. This was modified by adding polyacrylamide (PAA) with 2% whey protein isolate (WPI) or Tween 80 as foaming agents. Foaming was conducted in a stirred column. Rotation speed N and gas-to-liquid flow ratio (G/L) were varied. Overrun, average bubble size d (32), texture and stability were measured using densimetry, image analysis, and rheometry, respectively. Experimental results showed that 0.01% PAA did not modify the viscosity of 2% WPI models, but conferred low elastic behavior. PAA (0.05%) doubled matrix viscosity and drastically increased elasticity. The increase of elasticity became slower for further PAA addition. Foaming experiments demonstrated that theoretical overrun could not be achieved for inelastic WPI models in two cases: for high viscosity and low N, as dispersion effectiveness was reduced; for high G/L and N because of enhanced coalescence. Matrix elasticity was shown to increase overrun at constant viscosity for high G/L by enhancing interface stabilization. However, in elastic models, gas dispersion was more difficult and d (32) was higher than in inelastic fluids of similar viscosity. Finally, when the limiting step was dispersion, foaming was shown to be negatively affected by matrix elasticity.

Coalescence of droplets in viscoelastic matrix with diffuse interface under simple shear flow

AbstractThe coalescence process of two droplets in simple shear flow was modeled and simulated by the diffuse interface method. The collision between two droplets was investigated. The systems with small Peclet number, which denotes highly diffuse ability of concentration, were found to coalesce faster and easier due to the overlap of interfacial layers. The effect of matrix elasticity on droplet coalescence was studied thoroughly. The matrix elasticity was found to decrease the hydrodynamic interactions between droplets, and delay the coalescence process. © 2007 Wiley Periodicals, Inc. J Polym Sci Part B: Polym Phys 45: 1856–1869, 2007

Effect of Matrix Elasticity on Affinity Binding and Release of Bioparticles. Elution of Bound Cells by Temperature-Induced Shrinkage of the Smart Macroporous Hydrogel

The first step of bacterial or viral invasion is affinity and presumably multisite binding of bioparticles to an elastic matrix like a living tissue. We have demonstrated that model bioparticles such as inclusion bodies (spheres of about 1 microm in size) Escherichia coli cells (rods 1 x 3 microm), yeast cells (8 microm spheres), and synthetic microgel particles (0.4 microm spheres) are binding via different affinity interactions (IgG antibody-protein A, sugar-lectin, and metal ion-chelate) to a macroporous hydrogel (MH) matrix bearing appropriate ligands. The elastic deformation of the MH results in the detachment of affinity bound bioparticles. The particle detachment on elastic deformation is believed to be due to multipoint attachment of the particles to affinity matrix and the disturbance of the distance between affinity ligands when the matrix is deformed. No release of affinity bound protein occurred on elastic deformation. The efficiency of the particle release by the elastic deformation depends on the density of the ligands at the particle surface as well as on the elasticity of the matrix for relatively large particles. The release of the particles occurred irrespectively of whether the deformation was caused by external forces (mechanical deformation) or internal forces (the shrinkage of thermosensitive macroporous poly-N-isopropylacrylamide hydrogel on increase in temperature).

Break-up of a Newtonian drop in a viscoelastic matrix under simple shear flow

The effect of matrix elasticity on the break-up of an isolated Newtonian drop under step shear flow is herein presented. Constant-viscosity, elastic polymer solutions (Boger fluids) were used as matrix phase. Newtonian silicon oils were used as drop phase. Three viscosity ratios were explored (drop/matrix), i.e. 2, 0.6 and 0.04. Following the theoretical analysis of Greco [Greco F (2002) J Non-Newtonian Fluid Mech 107:111–131], the role of elasticity on drop fluid dynamics was quantified according to the value of the parameter p=τ/τem, where τ is a constitutive relaxation time of the matrix fluid and τem is the emulsion time. Different fluids were prepared in order to have p ranging from 0.1 to 10. At all the viscosity ratios explored, break-up was hindered by matrix elasticity. The start-up transient of drop deformation, at high, but sub-critical capillary numbers, showed an overshoot, during which the drop enhanced its orientation toward the flow direction. Both phenomena increase if the p parameter increases. Finally, the non-dimensional pinch-off length and break-up time were also found to increase with p.