Types Of Mechanical Stimuli Research Articles

Noncontact elasticity measurement of hydrogels in a culture dish using reverberant optical coherence elastography

Estimating the elasticity of hydrogel phantoms in a cell culture plane is important for understanding the cell behavior in response to various types of mechanical stimuli. Hence, a noncontact tool for measuring the elastic properties of hydrogel phantoms in such three-dimensional cell cultures is required. A well-known method to determine the mechanical properties of hydrogels is the transient wave method. However, due to the multiple reflections of waves from the boundaries, a bigger cell culture plane or multiple directional filters may be required. In this study, we utilized reverberant shear wave elastography, which is based on the autocorrelation principle, to evaluate the shear wave speed in hydrogel samples within a culture dish. Numerical simulations were performed first to confirm the validity of the reverberant elastography method. Subsequently, we used this method to measure the wave speeds in hydrogel phantoms with different concentrations. Shear rheology tests were also performed, and their results were found to be in good agreement with the measured shear wave speeds. The proposed method could be useful for measuring the elasticity of tissues in tissue engineering applications in an inexpensive and noncontact manner.

Active viscoelastic models for cell and tissue mechanics.

Living cells are out of equilibrium active materials. Cell-generated forces are transmitted across the cytoskeleton network and to the extracellular environment. These active force interactions shape cellular mechanical behaviour, trigger mechano-sensing, regulate cell adaptation to the microenvironment and can affect disease outcomes. In recent years, the mechanobiology community has witnessed the emergence of many experimental and theoretical approaches to study cells as mechanically active materials. In this review, we highlight recent advancements in incorporating active characteristics of cellular behaviour at different length scales into classic viscoelastic models by either adding an active tension-generating element or adjusting the resting length of an elastic element in the model. Summarizing the two groups of approaches, we will review the formulation and application of these models to understand cellular adaptation mechanisms in response to various types of mechanical stimuli, such as the effect of extracellular matrix properties and external loadings or deformations.

PKA mediates modality-specific modulation of the mechanically gated ion channel PIEZO2

PKA is a downstream effector of many inflammatory mediators that induce pain hypersensitivity by increasing the mechanosensitivity of nociceptive sensory afferent. Here, we examine the molecular mechanism underlying PKA-dependent modulation of the mechanically activated ion channel PIEZO2, which confers mechanosensitivity to many nociceptors. Using phosphorylation site prediction algorithms, we identified multiple putative and highly conserved PKA phosphorylation sites located on intracellular intrinsically disordered regions of PIEZO2. Site-directed mutagenesis and patch-clamp recordings showed that substitution of one or multiple putative PKA sites within a single intracellular domain does not alter PKA-induced PIEZO2 sensitization, whereas mutation of a combination of nine putative sites located on four different intracellular regions completely abolishes PKA-dependent PIEZO2 modulation, though it remains unclear whether all or just some of these nine sites are required. By demonstrating that PIEZO1 is not modulated by PKA, our data also reveal a previously unrecognized functional difference between PIEZO1 and PIEZO2. Moreover, by demonstrating that PKA only modulates PIEZO2 currents evoked by focal mechanical indentation of the cell, but not currents evoked by pressure-induced membrane stretch, we provide evidence suggesting that PIEZO2 is a polymodal mechanosensor that engages different protein domains for detecting different types of mechanical stimuli.

Mechanically resilient integrated electronics realized using interconnected 2D gold-nanosheet elastomeric electrodes

With the growing interest in wearable devices in recent decades, considerable effort has been devoted to developing mechanical elastomeric devices such as sensors, transistors, logic circuits, and integrated circuits. To successfully implement elastomeric devices subjected to large mechanical deformations or stretching, all the components, including conductors, semiconductors, and dielectrics, must have high stability and mechanical sustainability. Elastomeric conductors, which exhibit excellent electrical performances under mechanical deformations, are key components of elastomeric devices. Herein, we prepared fully elastomeric electrodes based on interconnected 2D gold nanosheets (AuNSs) to develop mechanically resilient integrated electronics. The AuNS elastomeric electrodes exhibited a sheet resistance of less than 2 Ω/sq under 50% stretching and sustained 100,000 stretching–releasing cycles. These electrodes with a dedicated design were used in combination with elastomeric semiconductors of P3HT nanofibrils in the PDMS elastomer (P3NF/PDMS) and an ion gel as a dielectric to realize elastomeric transistors, inverters, and NOR and NAND logic gates. Additionally, an elastomeric 8 × 8 transistor array that can sustain various types of mechanical stimuli was successfully demonstrated. Furthermore, the elastomeric electronic devices implemented on a soft robot showed no interfering performances during robot gripping motion. The proposed framework is expected to aid in the rapid development and broaden the application scope of soft electronics.

Intrinsically disordered intracellular domains control key features of the mechanically-gated ion channel PIEZO2

A central question in mechanobiology is how mechanical forces acting in or on cells are transmitted to mechanically-gated PIEZO channels that convert these forces into biochemical signals. Here we examined the role of the intracellular domains of PIEZO2, which account for 25% of the channel, and demonstrate that these domains fine-tune properties such as poking and stretch-sensitivity, velocity coding and single channel conductance. Moreover, we show that the intrinsically disordered linker between the transmembrane helices twelve and thirteen (IDR5) is required for the activation of PIEZO2 by cytoskeleton-transmitted forces. The deletion of IDR5 abolishes PIEZO2-mediated inhibition of neurite outgrowth, while it only partially affected its sensitivity to cell indentation and does not alter its stretch sensitivity. Thus, we propose that PIEZO2 is a polymodal mechanosensor that detects different types of mechanical stimuli via different force transmission pathways, which highlights the importance of utilizing multiple complementary assays when investigating PIEZO function.

The Individual and Combined Effects of Shear, Tension, and Flexure on Aortic Heart Valve Endothelial Cells in Culture

The necessity of living engineered heart valves to treat patients with severe heart disease poses a challenge to tissue engineers. To reach such goal it is crucial to fully understand the role and the activities of valvular endothelial cells (VECs) when they face different types of mechanical stimuli. This study focuses on decomposing the roles of different mechanical stimuli on heart valve endothelial surfaces and the response of VECs in terms of morphology and phenotype change. This study utilizes soft hydrogel-based scaffolds to use as a substrate for cell culture to mimic heart valve tissue leaflet. VECs were cultured as a monolayer on the gel surface and different types of mechanical stimuli were applied. Finally, the response of cells was investigated in terms of morphology and protein expression changes. Single stimuli introduces actin fibers reorganization in VECs, change in cell morphology, and higher mesenchymal protein expression. On the other hand, combined stimuli application has lower impact on actin fibers reorganization and cell morphology change, with lower mesenchymal protein expression. When VECs face a single mechanical stimuli, they undergo transdifferentiation and transform into mesenchymal cells. However, when these cells face a combination of mechanical stimuli, the rate of transformation decreases compared to single stimuli applications. This indicates that a single stimulus induces endothelial to mesenchymal transition in VECs while the process is slower under the combination of multiple mechanical stimuli.

Front Cover: Mechanochromic Luminescence and Solid‐State Circularly Polarized Luminescence of a Chiral Diamine‐Linked Bispyrene (ChemPhotoChem 10/2021)

The Front Cover illustrates the change in circularly polarized luminescence (CPL) of a chiral diamine-linked bispyrene caused by two types of mechanical stimuli, namely ultrasonication and grinding, analogous to the volcanic eruption and the collision of two volcanic rocks depicted in the image. More information can be found in the Article by Suguru Ito, Yoshitane Imai and co-workers.

Mechanochromic Luminescence and Solid‐State Circularly Polarized Luminescence of a Chiral Diamine‐Linked Bispyrene

AbstractDespite the recent increased interest in organic small molecule luminophores that exhibit circularly polarized luminescence (CPL) or mechanochromic luminescence (MCL), MCL behavior in chiral luminophores that display solid‐state CPL still remains elusive. Herein, it is reported that a chiral diamine‐linked bispyrene derivative exhibits a rare MCL behavior in response to two different types of mechanical stimuli, namely utrasonication and grinding. Moreover, an unprecedented switching of solid‐state CPL has been achieved for the chiral bispyrene between two states that can respond to the different mechanical stimuli. These findings should help to advance the development of practical CPL materials.

Rapid Prototyping of 3D Biochips for Cell Motility Studies Using Two-Photon Polymerization.

The study of cellular migration dynamics and strategies plays a relevant role in the understanding of both physiological and pathological processes. An important example could be the link between cancer cell motility and tumor evolution into metastatic stage. These strategies can be strongly influenced by the extracellular environment and the consequent mechanical constrains. In this framework, the possibility to study the behavior of single cells when subject to specific topological constraints could be an important tool in the hands of biologists. Two-photon polymerization is a sub-micrometric additive manufacturing technique that allows the fabrication of 3D structures in biocompatible resins, enabling the realization of ad hoc biochips for cell motility analyses, providing different types of mechanical stimuli. In our work, we present a new strategy for the realization of multilayer microfluidic lab-on-a-chip constructs for the study of cell motility which guarantees complete optical accessibility and the possibility to freely shape the migration area, to tailor it to the requirements of the specific cell type or experiment. The device includes a series of micro-constrictions that induce different types of mechanical stress on the cells during their migration. We show the realization of different possible geometries, in order to prove the versatility of the technique. As a proof of concept, we present the use of one of these devices for the study of the motility of murine neuronal cancer cells under high physical confinement, highlighting their peculiar migration mechanisms.

Mechanical property variation of AgNW/PDMS nanocomposites for fully elastomeric electrodes

Soft electronics necessitate the use of elastomeric components that can sustain dynamic mechanical strain for utilization in highly durable, reliable, and wearable applications. Notably, the development of elastomeric electrodes has been of significant interest for both academic and industrial research. High electrical conductivity, low performance discrepancy under various types of mechanical stimuli, and stable repeatability in the influence of dynamic strain cycles are considered to be some of the major prerequisites for the best soft electronics. Thus, a great deal of effort has been devoted to the formation of such elastomeric electrodes via special architecture and structural engineering. Recent remarkable outcomes in elastomeric electrodes have been accomplished by forming nanocomposites with a percolated network of low-dimensional metallic materials and an elastomer. This approach has aided in achieving scalable manufacturing, soluble printing, extreme mechanical stretchability, and low costs. Herein, we report nanocomposites of silver nanowires (AgNWs) and polydimethylsiloxane (PDMS) with different mechanical properties in order to maximize the electrical performance under various types of mechanical stimuli. Owing to the percolated AgNWs embedded in elastomeric PDMS, the resulting elastomeric electrodes can sustain a mechanical strain of up to 50%, while maintaining a resistance that is below 5 Ω/sq. In addition, a moderate increase in the sheet resistance was observed even after 130 k stretch-release cycles. Our AgNW elastomeric electrodes can be used in elastomeric heaters and have led to the successful capture of dynamic electrophysiological human bio-signals.

Paper-based in vitro tissue chip for delivering programmed mechanical stimuli of local compression and shear flow

Mechanical stimuli play important roles on the growth, development, and behavior of tissue. A simple and novel paper-based in vitro tissue chip was developed that can deliver two types of mechanical stimuli—local compression and shear flow—in a programmed manner. Rat vascular endothelial cells (RVECs) were patterned on collagen-coated nitrocellulose paper to create a tissue chip. Localized compression and shear flow were introduced by simply tapping and bending the paper chip in a programmed manner, utilizing an inexpensive servo motor controlled by an Arduino microcontroller and powered by batteries. All electrical compartments and a paper-based tissue chip were enclosed in a single 3D-printed enclosure, allowing the whole device to be independently placed within an incubator. This simple device effectively simulated in vivo conditions and induced successful RVEC migration in as early as 5 h. The developed device provides an inexpensive and flexible alternative for delivering mechanical stimuli to other in vitro tissue models.Graphical abstract

Mechanical stimuli enhance simultaneous differentiation into oesophageal cell lineages in a double-layered tubular scaffold.

The tissue-engineered oesophagus serves as an alternative and promising therapeutic approach for long-gap oesophageal replacement. This study proposes an advanced in vitro culture platform focused on construction of the oesophagus by combining an electrospun double-layered tubular scaffold, stem cells, biochemical reagents, and biomechanical factors. Human mesenchymal stem cells were seeded onto the inner and outer surfaces of the scaffold. Mechanical stimuli were applied with a hollow organ bioreactor along with different biochemical reagents inside and outside of the scaffold. Electrospun fibres in a tubular scaffold were found to be randomly and circumferentially oriented for the inner and outer surfaces, respectively. Amongst the two types of mechanical stimuli, the intermittent shear flow that can simultaneously cause circumferential stretching due to hydrostatic pressure, and shear stress caused by flow on the inner surface, was found to be more effective for simultaneous differentiation into epithelial and muscle lineage than steady shear flow. Under these conditions, the expression of epithelial markers on the inner surface was significantly observed, although it was minimal on the outer surface. Muscle differentiation showed the opposite expression pattern. Meanwhile, the mechanical tests showed that the strength of the scaffold was improved after incubation for 14days. We have developed a potential platform for tissue-engineered oesophagus construction. Specifically, simultaneous differentiation into epithelial and muscle lineages can be achieved by utilizing the double-layered scaffold and appropriate mechanical stimulation.

Self-organizing cell tactile perception which depends on mechanical stimulus history

Living organisms are capable of self-reproduction, self-repair, and high adaptability through self-organization. By applying biological materials such as cells and tissues to a robot, the resulting bio-machine hybrid system can perform important biological functions beyond those of traditional robots. In this study, we focused on the mechanobiological properties of a C2C12 cell, which is an immortalized mouse myoblast cell line. Calcium ion levels increased when the cells were subjected to mechanical stimuli. This type of physiological phenomenon can be applied in novel tactile sensors to develop a bio-machine hybrid system. We verified the tactile sensation characteristics of the cells. We cultured the cells on a thin polydimethylsulfoxane (PDMS) membrane. We applied two types of mechanical stimuli to opposite sides of the PDMS membrane. Next, we observed the cell distributions and quantified the increase in calcium ion levels inside the cells for tactile sensation. Our results demonstrate that the mechanical stimuli caused significant changes in the orientation and local density of cell aggregation. Interestingly, we confirmed that calcium ion increases depend on the history of mechanical stimuli application.

Termination of cell-type specification gene programs by the miR-183 cluster determines the population sizes of low-threshold mechanosensitive neurons.

Touch and mechanical sensations require the development of several different kinds of sensory neurons dedicated to respond to certain types of mechanical stimuli. The transcription factor Shox2 (short stature homeobox2) is involved in the generation of TRKB+ low-threshold mechanoreceptors (LTMRs), but mechanisms terminating this program and allowing alternative fates are unknown. Here, we show that the conditional loss of the miR-183-96-182 cluster in mouse leads to a failure of extinction of Shox2 during development and an increase in the proportion of Aδ LTMRs (TRKB+/NECAB2+) neurons at the expense of Aβ slowly adapting (SA)-LTMRs (TRKC+/Runx3-) neurons. Conversely, overexpression of miR-183 cluster that represses Shox2 expression, or loss of Shox2, both increase the Aβ SA-LTMRs population at the expense of Aδ LTMRs. Our results suggest that the miR-183 cluster determines the timing of Shox2 expression by direct targeting during development, and through this determines the population sizes of Aδ LTMRs and Aβ SA-LTMRs.

Bone Response to Physical Stimulus From In Vitro to In Vivo

Low-intensity pulsed ultrasound (LIPUS) is considered as a type of mechanical stimulus (mechanical stress). When the bone fracture part is treated with a low power ultrasonic pulse, various cells present in the fracture part act as a sensor of the mechanism. It has been reported that the acceptance of mechanical stress is mediated through the cell adhesion factor integrin at the cytoskeleton. Bone response to physical stimulus is different from acceptance of extension (compression) stimulus, which is well-known as mechanical stress acceptance. Accepted information is delivered intracellularly via various intracellular transmission systems and regulates the transcriptional regulatory region of the target gene. The reacted cells act on these stimuli to autocrine, paracrine, differentiation promoting action, cell migration action and the like. At the fracture site, the fracture healing process as wound healing is progressing, and various cell reactions occur simultaneously or overlapping in time and space. It is thought that when the low power ultrasonic pulse is irradiated, reaction by the above-mentioned mechanical stress occurs in each cell, thereby promoting the overall fracture healing process.

ALTERATIONS OF ECCENTRICALLY EVOKED PROTEIN SYNTHESIS IN ISOLATED RAT SOLEUS FOLLOWING GRAVITATIONAL UNLOADING: ROLE OF FAK

ALTERATIONS OF ECCENTRICALLY EVOKED PROTEIN SYNTHESIS IN ISOLATED RAT SOLEUS FOLLOWING GRAVITATIONAL UNLOADING: ROLE OF FAK

High‐Frequency Mechanostimulation of Cell Adhesion

AbstractCell adhesion is regulated by molecularly defined protein interactions and by mechanical forces, which can activate a dynamic restructuring of adhesion sites. Previous attempts to explore the response of cell adhesion to forces have been limited to applying mechanical stimuli that involve the cytoskeleton. In contrast, we here apply a new, oscillatory type of stimulus through push–pull azobenzenes. Push–pull azobenzenes perform a high‐frequency, molecular oscillation upon irradiation with visible light that has frequently been applied in polymer surface relief grating. We here use these oscillations to address single adhesion receptors. The effect of molecular oscillatory forces on cell adhesion has been analyzed using single‐cell force spectroscopy and gene expression studies. Our experiments demonstrate a reinforcement of cell adhesion as well as upregulated expression levels of adhesion‐associated genes as a result of the nanoscale “tickling” of integrins. This novel type of mechanical stimulus provides a previously unprecedented molecular control of cellular mechanosensing.

High-Frequency Mechanostimulation of Cell Adhesion.

Cell adhesion is regulated by molecularly defined protein interactions and by mechanical forces, which can activate a dynamic restructuring of adhesion sites. Previous attempts to explore the response of cell adhesion to forces have been limited to applying mechanical stimuli that involve the cytoskeleton. In contrast, we here apply a new, oscillatory type of stimulus through push–pull azobenzenes. Push–pull azobenzenes perform a high‐frequency, molecular oscillation upon irradiation with visible light that has frequently been applied in polymer surface relief grating. We here use these oscillations to address single adhesion receptors. The effect of molecular oscillatory forces on cell adhesion has been analyzed using single‐cell force spectroscopy and gene expression studies. Our experiments demonstrate a reinforcement of cell adhesion as well as upregulated expression levels of adhesion‐associated genes as a result of the nanoscale “tickling” of integrins. This novel type of mechanical stimulus provides a previously unprecedented molecular control of cellular mechanosensing.

A mathematical model of tissue-engineered cartilage development under cyclic compressive loading.

In this work a coupled model of solute transport and uptake, cell proliferation, extracellular matrix synthesis and remodeling of mechanical properties accounting for the impact of mechanical loading is presented as an advancement of a previously validated coupled model for free-swelling tissue-engineered cartilage cultures. Tissue-engineering constructs were modeled as biphasic with a linear elastic solid, and relevant intrinsic mechanical stimuli in the constructs were determined by numerical simulation for use as inputs of the coupled model. The mechanical dependent formulations were derived from a calibration and parametrization dataset and validated by comparison of normalized ratios of cell counts, total glycosaminoglycans and collagen after 24-h continuous cyclic unconfined compression from another dataset. The model successfully fit the calibration dataset and predicted the results from the validation dataset with good agreement, with average relative errors up to 3.1 and 4.3%, respectively. Temporal and spatial patterns determined for other model outputs were consistent with reported studies. The results suggest that the model describes the interaction between the simultaneous factors involved in in vitro tissue-engineered cartilage culture under dynamic loading. This approach could also be attractive for optimization of culture protocols, namely through the application to longer culture times and other types of mechanical stimuli.

Kinesin-Driven Active Substrate Giving Stochastic Mechanical Stimuli to Cells for Characterization.

We present a new platform to give stochastic mechanical stimuli to cells for their characterization. There nano- and micrometer scaled fluctuations are generated by an engineered motor protein system of kinesin-microtubules (MTs) on a solid surface. Cells have abilities to deform in many ways during homeostatic metabolism, tissue forming processes, cancer developments, and so on. Namely, cells in biological tissues are exposed to noise-like stochastic movements at nano- and micrometer-scales, which mainly come from the mechanical environment surrounding the cells. Although cells seem to have the potential to respond to such types of mechanical stimuli, the influences on cellular behaviors are poorly understood. As a first attempt to verify an effect of noise-like mechanical stimuli in vitro, we prepared a system to give stochastic mechanical stimuli to cells using a technique of in vitro motility assay for a kinesin-MT system. An active substrate was obtained by integrating movements of MTs on a kinesin-coated glass surface via cross-linkage, and stochastic mechanical stimuli at the cell-scale were successfully applied to the seeded cells. There, traveling distances of the cells over one cell length were observed until they started to adhere. When metastatic melanoma cells were exposed to the stochastic mechanical stimuli, unusually long protrusions or extensions of cell bodies were observed. Cellular aggregations were also promoted through the movements on this active substrate which could disturb the landing and enhance the collisions of the cells. This approach giving mechanical stimuli to cells in a stochastic manner at nano- and micrometer-scales might allow us to uncover unknown behaviors of cells, which might contribute to research fields requiring our understanding on the mechanical nature of cells, such as cancer diagnosis and regenerative medicine.