Wide Range Of Stiffness Research Articles

Peptide Domains as Reinforcement in Protein-Based Elastomers

Proteins are a widely available biomass source for synthesizing strong and tough engineering polymers because of their propensity to hydrogen bond, chemically stable amide backbone, and demonstrated efficacy at forming relevant material structures in nature. Because the properties of polypeptides in many ways mimic urethane bonds and hard domains, herein proteins are explored as the reinforcing component in a polyurethane-inspired elastomer. Materials are synthesized using a two-step process: first, protein is methacrylated, and then copolymerized with (meth)acrylate comonomers to link protein domains with rubbery polymer chains. This is demonstrated with water-soluble proteins, whey protein and β-lactoglobulin, and a comonomer, hydroxypropyl acrylate (HPA). The resulting elastomers are amorphous and disordered but have microphase-separated morphologies. Materials with a wide range of stiffnesses have been prepared by varying the fraction of protein macro-cross-linkers in the materials. The protein aggreg...

Video-tactile pneumatic sensor for soft tissue elastic modulus estimation

BackgroundA new sensor for estimating elasticity of soft tissues such as a liver was developed for minimally invasive surgery application.MethodsBy measuring deformation and adjusting internal pressure of the pneumatic sensor head, the sensor can be used to do palpation (indentation) of tissues with wide range of stiffness. A video camera installed within the sensor shell is used to register the radius of the contact area. Based on finite element model simulations and the measured data, elastic modulus of the indented soft tissue can be calculated.Results and conclusionsThree phantom materials, namely plastic, silicone and gelatin, with varied stiffness were tested. The experimental results demonstrated that the new sensor can obtain highly reliable data with error less than 5%. The new sensor might be served as an instrument in laparoscopic surgery for diagnosis of pathological tissues or internal organs.

Different TCR-induced T lymphocyte responses are potentiated by stiffness with variable sensitivity.

T cells are mechanosensitive but the effect of stiffness on their functions is still debated. We characterize herein how human primary CD4+ T cell functions are affected by stiffness within the physiological Young's modulus range of 0.5 kPa to 100 kPa. Stiffness modulates T lymphocyte migration and morphological changes induced by TCR/CD3 triggering. Stiffness also increases TCR-induced immune system, metabolism and cell-cycle-related genes. Yet, upon TCR/CD3 stimulation, while cytokine production increases within a wide range of stiffness, from hundreds of Pa to hundreds of kPa, T cell metabolic properties and cell cycle progression are only increased by the highest stiffness tested (100 kPa). Finally, mechanical properties of adherent antigen-presenting cells modulate cytokine production by T cells. Together, these results reveal that T cells discriminate between the wide range of stiffness values found in the body and adapt their responses accordingly.

Nonlinear mechanics of hybrid polymer networks that mimic the complex mechanical environment of cells

The mechanical properties of cells and the extracellular environment they reside in are governed by a complex interplay of biopolymers. These biopolymers, which possess a wide range of stiffnesses, self-assemble into fibrous composite networks such as the cytoskeleton and extracellular matrix. They interact with each other both physically and chemically to create a highly responsive and adaptive mechanical environment that stiffens when stressed or strained. Here we show that hybrid networks of a synthetic mimic of biological networks and either stiff, flexible and semi-flexible components, even very low concentrations of these added components, strongly affect the network stiffness and/or its strain-responsive character. The stiffness (persistence length) of the second network, its concentration and the interaction between the components are all parameters that can be used to tune the mechanics of the hybrids. The equivalence of these hybrids with biological composites is striking.

An Indirect Indentation Method for Evaluating the Linear Viscoelastic Properties of the Brain Tissue.

Indentation experiments offer a robust, fast, and repeatable testing method for evaluating the mechanical properties of the solid-state materials in a wide stiffness range. With the advantage of requiring a minimal sample preparation and multiple tests on a small piece of specimen, this method has recently become a popular technique for measuring the elastic properties of the biological materials, especially the brain tissue whose ultrasoft nature makes its mechanical characterization very challenging. Nevertheless, some limitations are associated with the indentation of the brain tissue, such as improper surface detection, negative initial contact force due to tip-tissue moisture interaction, and partial contact between the tip and the sample. In this study, an indirect indentation scheme is proposed to overcome the aforementioned difficulties. In this way, the indentation force is transferred from a sharp tip to the surface of the tissue slices via a rigid coverslip. To demonstrate the accuracy of this method, the linear viscoelastic properties of the white and gray matters of the bovine brain samples are measured by imposing small cyclic loads at different frequencies. The rate, regional, directional, and postmortem time dependence of the viscoelastic moduli are investigated and compared with the previous results from cyclic shear and monotonic experiments on the brain tissue. While findings of this research present a comprehensive set of information for the viscoelastic properties of the brain at a wide frequency range, the central goal of this paper is to introduce a novel experimentation technique with noticeable advantages for biomechanical characterization of the soft tissue.

Performance variation due to stiffness in a tuna-inspired flexible foil model

Tuna are fast, economical swimmers in part due to their stiff, high aspect ratio caudal fins and streamlined bodies. Previous studies using passive caudal fin models have suggested that while high aspect ratio tail shapes such as a tuna’s generally perform well, tail performance cannot be determined from shape alone. In this study, we analyzed the swimming performance of tuna-tail-shaped hydrofoils of a wide range of stiffnesses, heave amplitudes, and frequencies to determine how stiffness and kinematics affect multiple swimming performance parameters for a single foil shape. We then compared the foil models’ kinematics with published data from a live swimming tuna to determine how well the hydrofoil models could mimic fish kinematics. Foil kinematics over a wide range of motion programs generally showed a minimum lateral displacement at the narrowest part of the foil, and, immediately anterior to that, a local area of large lateral body displacement. These two kinematic patterns may enhance thrust in foils of intermediate stiffness. Stiffness and kinematics exhibited subtle interacting effects on hydrodynamic efficiency, with no one stiffness maximizing both thrust and efficiency. Foils of intermediate stiffnesses typically had the greatest coefficients of thrust at the highest heave amplitudes and frequencies. The comparison of foil kinematics with tuna kinematics showed that tuna motion is better approximated by a zero angle of attack foil motion program than by programs that do not incorporate pitch. These results indicate that open questions in biomechanics may be well served by foil models, given appropriate choice of model characteristics and control programs. Accurate replication of biological movements will require refinement of motion control programs and physical models, including the creation of models of variable stiffness.

Pressing movements and perceived force and displacement are influenced by object stiffness

Despite many previous studies on stiffness perception, few have investigated the exploratory procedures involved. This study evaluated whether stiffness range influences pressing movements and perception of force and displacement during stiffness discrimination tasks. Force and displacement data were obtained from 30 participants. Peak values of force and displacement, pressing duration and number of presses were analyzed. Two kinds of subjective evaluations were also recorded: perceived difference in force/displacement used to discriminate between specimens, and perceived effort. Although the number of presses and pressing duration were constant across a wide stiffness range, pressing strength was adjusted for the stiffness of objects, with harder specimens pressed more strongly. Further, even if the stiffnesses of two compared specimens were different, the pressing forces applied to the specimens approached the same magnitude at a higher stiffness range. Differences in force were most easily perceived at lower stiffness ranges, while displacement differences were perceived more readily at higher stiffness ranges. These results were consistent with those of previous studies. Finally, the reasons why stiffness range influenced pressing movements and perceived differences in force/displacement are discussed.

Custom Multiwell Plate Design for Rapid Assembly of Photopatterned Hydrogels.

The extracellular matrix provides both mechanical support and biochemical cues that influence cellular behavior. Matrix stiffness, in particular, has been found to regulate cellular morphology, motility, proliferation, differentiation, and drug responses among other behaviors. Thus, biomaterial platforms that exhibit wide range of stiffness and are available in a semi high-throughput format such as a multiwell plate would be useful for elucidating cell-substrate relationships. Polyacrylamide (PA) gels have been widely used as cell platforms since they span a range of stiffness between 0.3 and 300 kPa in Young's modulus, which encompasses all soft tissues. However, PA gels are time consuming and labor intensive to prepare, and are not amenable to a multiwell plate format. In this study, we present a novel custom multiwell plate design that allows for a one-step stiffness assay assembly that reduces preparation time and labor intensity by several fold. Gel stiffness is controlled by ultraviolet light intensity and exposure time to achieve a wide stiffness range from a single gel precursor solution. The geometry of the gels is defined by a custom photomask and gel thickness is controlled by spacers. A multiwell plate upper structure is designed similar to a regular multiwell plate such that a gel fits in each well and cells and media are added on top. The upper structure design allows for adequate gas exchange and minimum evaporation. Comparison between cell behaviors seeded in the custom and a standard multiwell plate demonstrated the suitability of the design as a cell culture platform. In summary, we describe and validate a novel custom design for an easy and rapid assembly of photopolymerizable PA-based stiffness assay.

Stretchable touch-sensing skin over padding for co-robots

For robots that work collaboratively with people, often referred to as ‘co-robots’, it would be beneficial for them to be soft or padded and to have a touch-sensing ‘skin’ to enable tactile environmental awareness. However, a sensing skin over a padding material that undergoes large deformations requires ‘stretchable’ materials, which may possess time-dependent or viscoelastic mechanical responses. In this work the roles that a padding layer plays when placed under a stretchable sensing layer was investigated. A strain-sensing skin was formed by coating a thin film of compliant piezoresistive sensing paint, consisting of exfoliated graphite in latex, onto a rubber membrane, and the response of the skin was characterized. The change in resistance was linear with tensile strain. The role of the padding material was then investigated under indentation by examining three foams and two elastomers. As expected, the padding enhanced energy dissipation as shown by hysteresis in the sensor response, which is linked to its protective function; the hysteresis was comparable for the five padding materials. The padding also provided an unexpected advantage: it magnified the change in resistance compared to that obtained under free displacement in air. While hysteresis in viscoelastic materials can largely be handled with an appropriate model, inconsistency cannot be, and the two elastomers were found to have unacceptably high variability because of micro-cracks and other defects in these materials. On the other hand, foams that had few defects and regular cell sizes gave good consistency across trials and different sensor positions over the padding. Combined with their lighter weight and availability in a wide range of stiffness, we conclude that foams make a better choice for padding of co-robots.

Consequences of Location-Dependent Organ of Corti Micro-Mechanics.

The cochlea performs frequency analysis and amplification of sounds. The graded stiffness of the basilar membrane along the cochlear length underlies the frequency-location relationship of the mammalian cochlea. The somatic motility of outer hair cell is central for cochlear amplification. Despite two to three orders of magnitude change in the basilar membrane stiffness, the force capacity of the outer hair cell’s somatic motility, is nearly invariant over the cochlear length. It is puzzling how actuators with a constant force capacity can operate under such a wide stiffness range. We hypothesize that the organ of Corti sets the mechanical conditions so that the outer hair cell’s somatic motility effectively interacts with the media of traveling waves—the basilar membrane and the tectorial membrane. To test this hypothesis, a computational model of the gerbil cochlea was developed that incorporates organ of Corti structural mechanics, cochlear fluid dynamics, and hair cell electro-physiology. The model simulations showed that the micro-mechanical responses of the organ of Corti are different along the cochlear length. For example, the top surface of the organ of Corti vibrated more than the bottom surface at the basal (high frequency) location, but the amplitude ratio was reversed at the apical (low frequency) location. Unlike the basilar membrane stiffness varying by a factor of 1700 along the cochlear length, the stiffness of the organ of Corti complex felt by the outer hair cell remained between 1.5 and 0.4 times the outer hair cell stiffness. The Y-shaped structure in the organ of Corti formed by outer hair cell, Deiters cell and its phalange was the primary determinant of the elastic reactance imposed on the outer hair cells. The stiffness and geometry of the Deiters cell and its phalange affected cochlear amplification differently depending on the location.

A new variable stiffness robot joint

Purpose – This study aims to propose a new variable stiffness robot joint (VSR-joint) for operating safely. More and more variable stiffness actuators are being designed and implemented because of their ability to minimize large forces due to shocks, to safely interact with the user and their ability to store and release energy in passive elastic elements. Design/methodology/approach – The design of VSR-joint is compact and integrated highly and the operating is simply. The mechanics, the principle of operation and the model of the VSR-joint are proposed. The principle of operation of VSR-joint is based on a lever arm mechanism with a continuously regulated pivot point. The VSR-joint features a highly dynamic stiffness adjustment along with a mechanically programmable system behavior. This allows an easy adaption to a big variety of tasks. Findings – Preliminary results are presented to demonstrate the fast stiffness regulation response and the wide range of stiffness achieved by the proposed VSR-joint design. Originality/value – In this paper, a new variable stiffness joint is proposed through changing the cantilever arm to change the performance of the elastic element, which is compact, small size and simple adjustment.

Crosstalk between focal adhesions and material mechanical properties governs cell mechanics and functions

Mechanical properties of materials strongly influence cell fate and functions. Focal adhesions are involved in the extremely important processes of mechanosensing and mechanotransduction. To address the relationship between the mechanical properties of cell substrates, focal adhesion/cytoskeleton assembly and cell functions, we investigated the behavior of NIH/3T3 cells over a wide range of stiffness (3–1000kPa) using two of the most common synthetic polymers for cell cultures: polyacrylamide and polydimethylsiloxane. An overlapping stiffness region was created between them to compare focal adhesion characteristics and cell functions, taking into account their different time-dependent behavior. Indeed, from a rheological point of view, polyacrylamide behaves like a strong gel (elastically), whereas polydimethylsiloxane like a viscoelastic solid. First, focal adhesion characteristics and dynamics were addressed in terms of material stiffness, then cell spreading area, migration rate and cell mechanical properties were correlated with focal adhesion size and assembly. Focal adhesion size was found to increase in the whole range of stiffness and to be in agreement in the overlapping rigidity region for the investigated materials. Cell mechanics directly correlated with focal adhesion lengths, whereas migration rate followed an inverse correlation. Cell spreading correlated with the substrate stiffness on polyacrylamide hydrogel, while no specific trend was found on polydimethylsiloxane. Substrate mechanics can be considered as a key physical cue that regulates focal adhesion assembly, which in turn governs important cellular properties and functions.

Simple polyacrylamide-based multiwell stiffness assay for the study of stiffness-dependent cell responses.

Currently, most of the in vitro cell research is performed on rigid tissue culture polystyrene (~1 GPa), while most cells in the body are attached to a matrix that is elastic and much softer (0.1-100 kPa). Since such stiffness mismatch greatly affects cell responses, there is a strong interest in developing hydrogel materials that span a wide range of stiffness to serve as cell substrates. Polyacrylamide gels, which are inexpensive and cover the stiffness range of all soft tissues in the body, are the hydrogel of choice for many research groups. However, polyacrylamide gel preparation is lengthy, tedious, and only suitable for small batches. Here, we describe an assay which by utilizing a permanent flexible plastic film as a structural support for the gels, enables the preparation of polyacrylamide gels in a multiwell plate format. The technique is faster, more efficient, and less costly than current methods and permits the preparation of gels of custom sizes not otherwise available. As it doesn't require any specialized equipment, the method could be easily adopted by any research laboratory and would be particularly useful in research focused on understanding stiffness-dependent cell responses.

Endothelial cell sensing, restructuring, and invasion in collagen hydrogel structures.

Experimental tools to model cell-tissue interactions will likely lead to new ways to both understand and treat cancer. While the mechanical properties and regulation of invasion have been recently studied for tumor cells, they have received less attention in the context of tumor vascular dynamics. In this article, we have investigated the interaction between the surfaces of structures encountered by endothelial cells invading their surrounding extracellular matrix (ECM) during angiogenesis. For this purpose, we have fabricated round and sharp geometries with various curvature and sharpness indices in collagen hydrogel over a wide range of stiffness to mimic different microenvironments varying from normal to tumor tissues. We have then cultured endothelial cells on these structures to investigate the bi-directional interaction between the cells and ECM. We have observed that cell invasion frequency is higher from the structures with the highest sharpness and curvature index, while interestingly the dependence of invasion on the local micro-geometry is strongest for the highest density matrices. Notably, structures with the highest invasion length are linked with higher deformation of side structures, which may be related to traction force-activated signaling suggesting further investigation. We have noted that round structures are more favorable for cell adhesion and in some cases round structures drive cell invasion faster than sharp ones. These results highlight the ability of endothelial cells to sense small variations in ECM geometry, and respond with a balance of matrix invasion as well as deformation, with potential implications for feedback mechanisms that may enhance vascular abnormality in response to tumor-induced ECM alterations.

Liquid contact resonance AFM: analytical models, experiments, and limitations

Contact resonance AFM (CR-AFM) is a scanning probe microscopy technique that utilizes the contact resonances of the AFM cantilever for concurrent imaging of topography and surface stiffness. The technique has not been used in liquid until recently due to analytical and experimental difficulties, associated with viscous damping of cantilever vibrations and fluid loading effects. To address these difficulties, (i) an analytical approach for contact resonances in liquid is developed, and (ii) direct excitation of the contact resonances is demonstrated by actuating the cantilever directly in a magnetic field. By implementing the analytical approach and the direct actuation through magnetic particles, quantitative stiffness imaging on surfaces with a wide range of stiffness can be achieved in liquid with soft cantilevers and low contact forces.

Mechanosensing of cells in 3D gel matrices based on natural and synthetic materials.

Cells in vivo typically are found in 3D matrices, the mechanical stiffness of which is important to the cell and tissue-scale biological processes. Although it is well characterized that as to how cells sense matrix stiffness in 2D substrates, the scenario in 3D matrices needs to be explored. Thus, materials that can mimic native 3D environments and possess wide, physiologically relevant elasticity are highly desirable. Natural polymer-based materials and synthetic hydrogels could provide an better 3D platforms to investigate the mechano-response of cells with stiffness comparable to their native environments. However, the limited stiffness range together with interdependence of matrix stiffness and adhesive ligand density are inherent in many kinds of materials, and hinder efforts to demonstrate the true effects contributed by matrix stiffness. These problems have been addressed by the recently emerging exquisitely designed materials based on native matrix components, designer matrices, and synthetic polymers. In this review, a variety of materials with a wide stiffness range that mimic the mechanical environment of native 3D matrices and the independent affection of stiffness for cellular behavior and tissue-level processes are discussed.

Zero displacement microelectromechanical force sensor using feedback control

Conventional microscale force sensors use moving parts to infer applied forces. Whenever physical deformations are involved, the sensor characteristics become a function of mechanical parameters, and there is an inevitable trade-off between the sensitivity and measurement range. We developed a microfabricated force sensor that uses feedback control to nullify any displacements within the device, directly transducing forces as high as 1.5 mN with a 7.8 nN resolution. The range and sensitivity of the device no longer depend on mechanical parameters, which allow the same device to be used to test samples with a wide range of stiffnesses without loss of accuracy.

Analysis of cracking resistance of bituminous mixtures using Fenix test

The aim of this study is to characterize the cracking resistance of several bituminous mixtures using Fénix test at different temperatures (from 30°C to −30°C) in order to minimize the phenomenon of reflective cracking in pavements. This direct tensile strength test evaluates the cracking resistance of asphalt mixes by calculating the energy dissipated during mixture cracking.Different types of bituminous mixtures containing different types of bitumen to obtain a wide stiffness range were analyzed. Environmental and sustainability aspects were considered when selecting the different bitumens (low temperature, rejuvenating agents, crumb rubber, among others).The mix with high performance polymer modified bitumen, with the highest energy curve, has the best response to cracking, especially in the temperature range between 5 and −10°C. Fatigue laws obtained through 4-point bending beam test confirm results. The mixes with the best response to cracking, under similar stiffness levels, have higher fracture energy.

Reprogramming cellular phenotype by soft collagen gels.

A variety of cell types exhibit phenotype changes in response to the mechanical stiffness of the substrate. Many cells excluding neurons display an increase in the spread area, actin stress fiber formation and larger focal adhesion complexes as substrate stiffness increases in a sparsely populated culture. Cell proliferation is also known to directly correlate with these phenotype changes/changes in substrate stiffness. Augmented spreading and proliferation on stiffer substrates require nuclear transcriptional regulator YAP (Yes associated protein) localization in the cell nucleus and is tightly coupled to larger traction force generation. In this study, we show that different types of fibroblasts can exhibit spread morphology, well defined actin stress fibers, and larger focal adhesions even on very soft collagen gels (modulus in hundreds of Pascals) as if they are on hard glass substrates (modulus in GPa, several orders of magnitude higher). Strikingly, we show, for the first time, that augmented spreading and other hard substrate cytoskeleton architectures on soft collagen gels are not correlated with the cell proliferation pattern and do not require YAP localization in the cell nucleus. Finally, we examine the response of human colon carcinoma (HCT-8) cells on soft collagen gels. Recent studies show that human colon carcinoma (HCT-8) cells form multicellular clusters by 2-3 days when cultured on soft polyacrylamide (PA) gels with a wide range of stiffness (0.5-50 kPa) and coated with an extracellular matrix, ECM (collagen monomer/fibronectin). These clusters show limited spreading/wetting on PA gels, form 3D structures at the edges, and eventually display a remarkable, dissociative metastasis like phenotype (MLP), i.e., epithelial to rounded morphological transition after a week of culture on PA gels only, but not on collagen monomer coated stiff polystyrene/glass where they exhibit enhanced wetting and form confluent monolayers. Here, we show that HCT-8 cell clusters also show augmented spreading/wetting on soft collagen gels and eventually form confluent monolayers as on rigid glass substrates and MLP is completely inhibited on soft collagen gels. Overall, these results suggest that cell-material interactions (soft collagen gels in this case) can induce cellular phenotype and cytoskeleton organization in a remarkably distinct manner compared to a classical synthetic polyacrylamide (PA) hydrogel cell culture model and may contribute in designing new functional biomaterials.

Mechanical Characterization of Human Monocyte Derived Antigen Presenting Cells

T cells are at the root of the immune response protecting living organisms against pathogens and tumor cells, and have recently been shown to respond to mechanical changes. The activation of T cells triggering the immune response requires the formation of cell-cell interaction between T lymphocytes and various antigen presenting cells (APC) (tumor cells, dendritic cells, macrophages, B lymphocytes...). Yet, little is known about the rigidity of these different APCs. In the present study we use the single-cell microplates assay to measure the mechanical properties of human monocyte derived APCs. We show that APCs present a wide range of stiffnesses, that are affected during differentiation, maturation and in response to inflammatory signals.Noteworthily, dendritic cells, the most efficient APC for T cell activation, present the closest rigidity to T cell rigidity. These results suggest that cell stiffness could be an important factor in immune cell-cell interactions and T cell mediated responses. We will then show preliminary studies on the role of rigidity in T cell activation ; involving laying T cells on substrates mimicking APCs rigidities and monitoring T cell activation and T cell / substrate interface organization.View Large Image | View Hi-Res Image | Download PowerPoint Slide