Effects Of Mechanical Stimuli Research Articles

Bone and Joint-on-Chip Platforms: Construction Strategies and Applications.

Organ-on-a-chip, also known as "tissue chip," is an advanced platform based on microfluidic systems for constructing miniature organ models in vitro. They can replicate the complex physiological and pathological responses of human organs. In recent years, the development of bone and joint-on-chip platforms aims to simulate the complex physiological and pathological processes occurring in human bones and joints, including cell-cell interactions, the interplay of various biochemical factors, the effects of mechanical stimuli, and the intricate connections between multiple organs. In the future, bone and joint-on-chip platforms will integrate the advantages of multiple disciplines, bringing more possibilities for exploring disease mechanisms, drug screening, and personalized medicine. This review explores the construction and application of Organ-on-a-chip technology in bone and joint disease research, proposes a modular construction concept, and discusses the new opportunities and future challenges in the construction and application of bone and joint-on-chip platforms.

Effect of Mechanical Stimuli on the Phenotypic Plasticity of Induced Pluripotent Stem-Cell-Derived Vascular Smooth Muscle Cells in a 3D Hydrogel.

Introduction: Vascular smooth muscle cells (VSMCs) play a pivotal role in vascular homeostasis, with dysregulation leading to vascular complications. Human-induced pluripotent stem-cell (hiPSC)-derived VSMCs offer prospects for personalized disease modeling and regenerative strategies. Current research lacks comparative studies on the impact of three-dimensional (3D) substrate properties under cyclic strain on phenotypic adaptation in hiPSC-derived VSMCs. Here, we aim to investigate the impact of intrinsic substrate properties, such as the hydrogel's elastic modulus and cross-linking density in a 3D static and dynamic environment, on the phenotypical adaptation of human mural cells derived from hiPSC-derived organoids (ODMCs), compared to aortic VSMCs. Methods and results: ODMCs were cultured in two-dimensional (2D) conditions with synthetic or contractile differentiation medium or in 3D Gelatin Methacryloyl (GelMa) substrates with varying degrees of functionalization and percentages to modulate Young's modulus and cross-linking density. Cells in 3D substrates were exposed to cyclic, unidirectional strain. Phenotype characterization was conducted using specific markers through immunofluorescence and gene expression analysis. Under static 2D culture, ODMCs derived from hiPSCs exhibited a VSMC phenotype, expressing key mural markers, and demonstrated a level of phenotypic plasticity similar to primary human VSMCs. In static 3D culture, a substrate with a higher Young's modulus and cross-linking density promoted a contractile phenotype in ODMCs and VSMCs. Dynamic stimulation in the 3D substrate promoted a switch toward a contractile phenotype in both cell types. Conclusion: Our study demonstrates phenotypic plasticity of human ODMCs in response to 2D biological and 3D mechanical stimuli that equals that of primary human VSMCs. These findings may contribute to the advancement of tailored approaches for vascular disease modeling and regenerative strategies.

Extracellular matrix cues regulate the differentiation of pluripotent stem cell-derived endothelial cells.

The generation of endothelial cells (ECs) from human pluripotent stem cells (PSCs) has been a promising approach for treating cardiovascular diseases for several years. Human PSCs, particularly induced pluripotent stem cells (iPSCs), are an attractive source of ECs for cell therapy. Although there is a diversity of methods for endothelial cell differentiation using biochemical factors, such as small molecules and cytokines, the efficiency of EC production varies depending on the type and dose of biochemical factors. Moreover, the protocols in which most EC differentiation studies have been performed were in very unphysiological conditions that do not reflect the microenvironment of native tissue. The microenvironment surrounding stem cells exerts variable biochemical and biomechanical stimuli that can affect stem cell differentiation and behavior. The stiffness and components of the extracellular microenvironment are critical inducers of stem cell behavior and fate specification by sensing the extracellular matrix (ECM) cues, adjusting the cytoskeleton tension, and delivering external signals to the nucleus. Differentiation of stem cells into ECs using a cocktail of biochemical factors has been performed for decades. However, the effects of mechanical stimuli on endothelial cell differentiation remain poorly understood. This review provides an overview of the methods used to differentiate ECs from stem cells by chemical and mechanical stimuli. We also propose the possibility of a novel EC differentiation strategy using a synthetic and natural extracellular matrix.

Effect of Mechanical Stimuli and Zoledronic Acid on the Femoral Bone Morphology in Rats with Obesity and Limited Mobility.

Our study aimed to compare the impact of zoledronic acid and whole-body vibration (WBV) as a non-pharmacological method of treatment for early obesity/immobility-related osteoporosis in male rat models. In total, 36 male Wistar rats were assigned to the following groups: obese control with immobility (Control, n = 12) and two experimental groups (n = 12 each), including obese and immobile rats subjected to whole-body vibration with an acceleration level of 3 m/s2 g (obesity and immobility + WBV) and obese and immobile rats that received an intramuscular injection of zoledronic acid at a dose of 0.025 mg/kg (obesity and immobility + ZOL). After the 8th and 16th week of treatment, n = 6 rats from each group were euthanized and isolated femora were subjected to a histological examination of bone, and analysis of the expression of osteoprotegerin (OPG) and the receptor activator of nuclear factor kappa-B ligand (RANKL) involved in bone turnover and the amount of thin collagen fibers (PSR stain). The obtained results showed that short-term vibrotherapy (up to 8 weeks) can lead to improvement in bone remodeling in rat models with obesity and limited mobility.

Using a meshless method to assess the effect of mechanical loading in angiogenesis

Although it is known that blood vessels can be found in mechanically active environments, less is known about the effect of mechanical stimulus in angiogenesis. Therefore, understanding how endothelial cells respond to a mechanical stimulus is essential to improve tissue vascularization and to promote wound healing and tissue engineering development. In this work, a meshless method is used to combine an elasticity formulation with a capillary growth algorithm. The final numerical model is capable to simulate the effect of compressive loading in angiogenesis, using three strain magnitudes (5, 10 and 30% strain). In this proposed model, the vascular endothelial growth factor gradient regulates the endothelial cell migration and the compressive loading affects the branching process. The numerical results showed that all the compressive loadings tested increased the vascular network length and the number of branches, being 5% strain magnitude the most effective one. The capillary network obtained resembles the one presented in experimental assays and the obtained numerical results coincided to the experimental ones. Nevertheless, this study possesses some limitations since the viscoelastic properties of the tissue, the dynamic loading effect and the effect of the time variable were not considered. In the future, the combination of computational and experimental studies will be very useful to understand and to define which are the mechanical cues that promote angiogenesis, allowing to improve tissue vascularization and, consequently, the wound healing process.

The Mechanosensory Role of Osteocytes and Implications for Bone Health and Disease States.

Bone homeostasis is a dynamic equilibrium between bone-forming osteoblasts and bone-resorbing osteoclasts. This process is primarily controlled by the most abundant and mechanosensitive bone cells, osteocytes, that reside individually, within chambers of porous hydroxyapatite bone matrix. Recent studies have unveiled additional functional roles for osteocytes in directly contributing to local matrix regulation as well as systemic roles through endocrine functions by communicating with distant organs such as the kidney. Osteocyte function is governed largely by both biochemical signaling and the mechanical stimuli exerted on bone. Mechanical stimulation is required to maintain bone health whilst aging and reduced level of loading are known to result in bone loss. To date, both in vivo and in vitro approaches have been established to answer important questions such as the effect of mechanical stimuli, the mechanosensors involved, and the mechanosensitive signaling pathways in osteocytes. However, our understanding of osteocyte mechanotransduction has been limited due to the technical challenges of working with these cells since they are individually embedded within the hard hydroxyapatite bone matrix. This review highlights the current knowledge of the osteocyte functional role in maintaining bone health and the key regulatory pathways of these mechanosensitive cells. Finally, we elaborate on the current therapeutic opportunities offered by existing treatments and the potential for targeting osteocyte-directed signaling.

Degradation of 3D-Printed Porous Polylactic Acid Scaffolds Under Mechanical Stimulus.

Polylactic acid (PLA) is a biodegradable polymer commonly used as a scaffold material to repair tissue defects, and its degradation is associated with mechanical stimulus. In this study, the effect of mechanical stimulus on the degradation of 3D-printed PLA scaffolds was investigated by in vitro experiments and an author-developed numerical model. Forty-five samples with porosity 64.8% were printed to carry out the degradation experiment within 90 days. Statistical analyses of the mass, volume fraction, Young’s modulus, and number average molecular weight were made, and the in vitro experiments were further used to verify the proposed numerical model of the scaffold degradation. The results indicated that the mechanical stimulus accelerated the degradation of the PLA scaffold, and the higher mechanical stimulus led to a faster degradation of the scaffolds at the late stage of the degradation process. In addition, the Young’s modulus and the normalized number average molecular weight of the PLA scaffolds between the experiments and the numerical simulations were comparable, especially for the number average molecular weight. The present study could be helpful in the design of the biodegradable PLA scaffolds.

Effect of cyclic mechanical loading on immunoinflammatory microenvironment in biofabricating hydroxyapatite scaffold for bone regeneration

It has been proven that the mechanical microenvironment can impact the differentiation of mesenchymal stem cells (MSCs). However, the effect of mechanical stimuli in biofabricating hydroxyapatite scaffolds on the inflammatory response of MSCs remains unclear. This study aimed to investigate the effect of mechanical loading on the inflammatory response of MSCs seeded on scaffolds. Cyclic mechanical loading was applied to biofabricate the cell-scaffold composite for 15 min/day over 7, 14, or 21 days. At the predetermined time points, culture supernatant was collected for inflammatory mediator detection, and gene expression was analyzed by qRT-PCR. The results showed that the expression of inflammatory mediators (IL1B and IL8) was downregulated (p < 0.05) and the expression of ALP (p < 0.01) and COL1A1 (p < 0.05) was upregulated under mechanical loading. The cell-scaffold composites biofabricated with or without mechanical loading were freeze-dried to prepare extracellular matrix-based scaffolds (ECM-based scaffolds). Murine macrophages were seeded on the ECM-based scaffolds to evaluate their polarization. The ECM-based scaffolds that were biofabricated with mechanical loading before freeze-drying enhanced the expression of M2 polarization-related biomarkers (Arginase 1 and Mrc1, p < 0.05) of macrophages in vitro and increased bone volume/total volume ratio in vivo. Overall, these findings demonstrated that mechanical loading could dually modulate the inflammatory responses and osteogenic differentiation of MSCs. Besides, the ECM-based scaffolds that were biofabricated with mechanical loading before freeze-drying facilitated the M2 polarization of macrophages in vitro and bone regeneration in vivo. Mechanical loading may be a promising biofabrication strategy for bone biomaterials.

Effects of Mechanical Stimulus on Beat Sequence of Cardiomyocytes with Feedback Control

Not only electrical stimulus but also direct mechanical stimulus induces beating of cardiac cells, particularly where the deformation by the stimulus is resemble to that created by beating neighboring cardiac cells. As primary cardiac cells forming aggregation become synchronized their beating while incubated more than 24 h, periodic stimulus artificially deforming cell aggregation like beat contraction by a probe may induce beating as pacemaker cell does. We fabricated an instrumentation to apply such periodic stimulus by a tungsten probe to a cardiac cell aggregate on a plastic Petri dish or on a PDMS substrate.

Micro-Vibration Stimulation Promotes M2 Polarization of Macrophages Beneficial for Osteogenesis Via Activating FAK/P38MAPK and Suppressing Ca &lt;sup&gt;2+&lt;/sup&gt;/ERK/ p65NF-κB Signaling Pathways

The emergence of osteoimmunology has demonstrated that bone system is closely related to immune system. However, most studies have investigated the interaction between immune cells and bone-related cells under a static cell culture, the effects of mechanical stimuli on the inflammatory responses of macrophages remain elusive. In this work, micro-vibration stimulation (MVS) was introduced into regulating macrophage polarization and functional status. The results found that MVS with 40 Hz in frequency and 0.3 g in magnitude could significantly raise the fraction of CD206-positive M2 population, up-regulate the expression of M2 marker genes, and enhance the secretion of M2 marker proteins in RAW 264.7 macrophages. It suggested that calcium ion and focal adhesion played an important role in transmitting MVS signal. Further inhibitor studies and western blotting analyses indicated that MVS-induced M2 polarization might be attributed to the activation of p38MAPK signaling and suppression of the ERK1/2 and p65NF-κB signaling. Moreover, MVS-mediated macrophage secretion enhanced osteoblastic differentiation of MC3T3-E1 pre-osteoblasts via Wnt/β-catenin and ERK1/2 signaling in a paracrine way. These findings highlight a critical role of MVS in inducing M2 polarization of macrophage to create a pro-osteogenic environment, which offers a drug-free modulatory strategy to regulate immune response and promote osteogenesis.

Effects of mechanical stimuli on structure and organization of bone nanocomposites in rats with glucocorticoid-induced osteoporosis.

Objective. Clinical use of glucocorticoids is a frequent cause of secondary osteoporosis, which reduces the mineral density of bones and results in pathological fractures. Mechanical stimulation as non-physiological high-frequency vibration with low acceleration prevents the loss of a crystalline component and stimulates the anabolic remodeling of the bone. The aim of the present research was to assess the impact of mechanical vibration on the bone structure in rats, which received glucocorticoids.Methods. Wistar rats were randomized into three groups: Vehicle control (Veh), Methylprednisolone sodium succinate (Mps), and Mps combined with whole-body vibration (WBV). Rats of Mps+WBV and Mps groups received 3 mg/kg/day of methylprednisolone every other day for 24 weeks and rats of Veh group received 0.9% saline (sodium chloride). The group of rats Mps+WBV was subjected to WBV for 30 minutes per day for five days a week with parameters 0.3 g and frequency 50 Hz. Relative amount of crystalline component and collagen in the bones was determined by X-ray diffraction (XRD) and calcium level - by atomic absorption spectroscopy. Bone tissue metabolism was assessed by determining the concentration of markers, in particular osteocalcin and Tartrate-resistant acid phosphatase (TRAP5b).Results. Glucocorticoids induced a considerable increase in the rats body mass (+13%) and decreased the content of mineral component in the femoral neck (-17%) in Mps group compared with Veh. The process of the bone metabolism was significantly accelerated, which is proven by an increased level of remodeling markers. It should be mentioned that WBV did not allow significant decrease in mineral component of the bone to 16th week of the experiment compared with Mps group, although these parameters did not achieve the indices in the Vehicle control group (-10%). Our investigation allows to suggest that mechanical high-frequency vibration of low intensity can partially inhibit the harmful consequences of glucocorticoids on bone structure in rats. Despite the positive impact of vibration on the bone tissue after Mps introduction in the 8th-16th week, this influence was not statistically reliable in the 24th week of the experiment.Conclusions. The results of our investigation on animal model indicate that non-physiological vertical mechanical vibrations are an effective means to prevent loss of a mineral bone component during treatment with glucocorticoids.

Effect of mechanical stimuli on remodeling and mineral density of the tibia in ovarioectomized rats

Effect of mechanical stimuli on remodeling and mineral density of the tibia in ovarioectomized rats

Enhanced junctional epithelial permeability in TRPV4-deficient mice.

Background and ObjectiveAs the interface between the oral cavity and the teeth, the junctional epithelial barrier is critical for gingival defense. The junctional epithelium is subject to mechanical stresses from biting force or external insults such as bacterial attacks, but little is known about the effects of mechanical stimuli on epithelial functions. Transient receptor potential vanilloid 4 (TRPV4) functions as a mechanosensitive nonselective cation channel. In the present study, based on marked expression of TRPV4 in the mouse junctional epithelium, we aimed to clarify the putative links between TRPV4 and junctional complexes in the junctional epithelium.Methods and ResultsHistological observations revealed that the junctional epithelium in TRPV4‐deficient (TRPV4−/−) mice had wider intercellular spaces than that in wild‐type (TRPV4+/+) mice. Exogenous tracer penetration in the junctional epithelium was greater in TRPV4−/− mice than in TRPV4+/+ mice, and immunoreactivity for adherens junction proteins was suppressed in TRPV4−/− mice compared with TRPV4+/+ mice. Analysis of a mouse periodontitis model showed greater bone volume loss in TRPV4−/− mice compared with TRPV4+/+ mice, indicating that an epithelial barrier deficiency in TRPV4−/− mice may be associated with periodontal complications.ConclusionThe present findings identify a crucial role for TRPV4 in the formation of adherens junctions in the junctional epithelium, which could regulate its permeability. TRPV4 may be a candidate pharmacological target to combat periodontal diseases.

Where No Hand Has Gone Before: Probing Mechanobiology at the Cellular Level.

Physical forces and other mechanical stimuli are fundamental regulators of cell behavior and function. Cells are also biomechanically competent: they generate forces to migrate, contract, remodel, and sense their environment. As the knowledge of the mechanisms of mechanobiology increases, the need to resolve and probe increasingly small scales calls for novel technologies to mechanically manipulate cells, examine forces exerted by cells, and characterize cellular biomechanics. Here, we review novel methods to quantify cellular force generation, measure cell mechanical properties, and exert localized piconewton and nanonewton forces on cells, receptors, and proteins. The combination of these technologies will provide further insight on the effect of mechanical stimuli on cells and the mechanisms that convert these stimuli into biochemical and biomechanical activity.

Dental Pulp Stem Cell Mechanoresponsiveness: Effects of Mechanical Stimuli on Dental Pulp Stem Cell Behavior.

Dental pulp is known to be an accessible and important source of multipotent mesenchymal progenitor cells termed dental pulp stem cells (DPSCs). DPSCs can differentiate into odontoblast-like cells and maintain pulp homeostasis by the formation of new dentin which protects the underlying pulp. DPSCs similar to other mesenchymal stem cells (MSCs) reside in a niche, a complex microenvironment consisting of an extracellular matrix, other local cell types and biochemical stimuli that influence the decision between stem cell (SC) self-renewal and differentiation. In addition to biochemical factors, mechanical factors are increasingly recognized as key regulators in DPSC behavior and function. Thus, microenvironments can significantly influence the role and differentiation of DPSCs through a combination of factors which are biochemical, biomechanical and biophysical in nature. Under in vitro conditions, it has been shown that DPSCs are sensitive to different types of force, such as uniaxial mechanical stretch, cyclic tensile strain, pulsating fluid flow, low-intensity pulsed ultrasound as well as being responsive to biomechanical cues presented in the form of micro- and nano-scale surface topographies. To understand how DPSCs sense and respond to the mechanics of their microenvironments, it is essential to determine how these cells convert mechanical and physical stimuli into function, including lineage specification. This review therefore covers some aspects of DPSC mechanoresponsivity with an emphasis on the factors that influence their behavior. An in-depth understanding of the physical environment that influence DPSC fate is necessary to improve the outcome of their therapeutic application for tissue regeneration.

Interstitial flow promotes macrophage polarization toward an M2 phenotype.

Tumor tissues are characterized by an elevated interstitial fluid flow from the tumor to the surrounding stroma. Macrophages in the tumor microenvironment are key contributors to tumor progression. While it is well established that chemical stimuli within the tumor tissues can alter macrophage behaviors, the effects of mechanical stimuli, especially the flow of interstitial fluid in the tumor microenvironment, on macrophage phenotypes have not been explored. Here, we used three-dimensional biomimetic models to reveal that macrophages can sense and respond to pathophysiological levels of interstitial fluid flow reported in tumors (∼3 µm/s). Specifically, interstitial flow (IF) polarizes macrophages toward an M2-like phenotype via integrin/Src-mediated mechanotransduction pathways involving STAT3/6. Consistent with this flow-induced M2 polarization, macrophages treated with IF migrate faster and have an enhanced ability to promote cancer cell migration. Moreover, IF directs macrophages to migrate against the flow. Since IF emanates from the tumor to the surrounding stromal tissues, our results suggest that IF could not only induce M2 polarization of macrophages but also recruit these M2 macrophages toward the tumor masses, contributing to cancer cell invasion and tumor progression. Collectively, our study reveals that IF could be a critical regulator of tumor immune environment.

Effects of Mechanical Stimuli on Profilin- and Formin-Mediated Actin Polymerization.

Self-assembling actin filaments not only form the basis of the cytoskeleton network in cells but also are utilized as nanosized building blocks to make novel active matter in which the dynamic polymerization and depolymerization of actin filaments play a key role. Formins belong to a main family of actin nucleation factors that bind to the barbed end of actin filaments and regulate actin polymerization through an interaction with profilin. Due to actomyosin contractility and relative rotation between formin and actin filaments, formin-dependent actin polymerization is subject to force and rotation constraints. However, it remains unclear how force and rotation constraints affect formin-dependent actin polymerization in the presence of profilin. Here, we show that for rotation-unconstrained actin filaments, elongation is accelerated by both force and profilin. The combined effect leads to surprisingly fast actin elongation that can approach the diffusion-limited rate at forces of a few piconewtons. The elongation of rotation-constrained filaments is also accelerated by profilin but is insensitive to applied force. We show that FH2, the main actin binding domain, plays the primary mechanosensing role. Together, the findings not only significantly advance our understanding of the mechanochemical regulation of formin-mediated actin polymerization in cells but also can potentially be utilized to make novel actin-based active matter.

Bone regeneration capacity of magnesium phosphate cements in a large animal model

Magnesium phosphate minerals have captured increasing attention during the past years as suitable alternatives for calcium phosphate bone replacement materials. Here, we investigated the degradation and bone regeneration capacity of experimental struvite (MgNH4PO4·6H2O) forming magnesium phosphate cements in two different orthotopic ovine implantation models. Cements formed at powder to liquid ratios (PLR) of 2.0 and 3.0 g ml−1 were implanted into trabecular bone using a non-load-bearing femoral drill-hole model and a load-bearing tibial defect model. After 4, 7 and 10 months the implants were retrieved and cement degradation and new bone formation was analyzed by micro-computed tomography (µCT) and histomorphometry. The results showed cement degradation in concert with new bone formation at both defect locations. Both cements were almost completely degraded after 10 months. The struvite cement formed with a PLR of 2.0 g ml−1 exhibited a slightly accelerated degradation kinetics compared to the cement with a PLR of 3.0 g ml−1. Tartrat-resistant acid phosphatase (TRAP) staining indicated osteoclastic resorption at the cement surface. Energy dispersive X-ray analysis (EDX) revealed that small residual cement particles were mostly accumulated in the bone marrow in between newly formed bone trabeculae. Mechanical loading did not significantly increase bone formation associated with cement degradation. Concluding, struvite-forming cements might be promising bone replacement materials due to their good degradation which is coupled with new bone formation. Statement of SignificanceRecently, the interest in magnesium phosphate cements (MPC) for bone substitution increased, as they exhibit high initial strength, comparably elevated degradation potential and the release of valuable magnesium ions. However, only few in vivo studies, mostly including non-load-bearing defects in small animals, have been performed to analyze the degradation and regeneration capability of MPC derived compounds. The present study examined the in vivo behavior of magnesiumammoniumphosphate hexahydrate (struvite) implants with different porosity in both mechanically loaded and non-loaded defects of merino sheep. For the first time, the effect of mechanical stimuli on the biological outcome of this clinically relevant replacement material is shown and directly compared to the conventional unloaded defect situation in a large animal model.

Influence of Extracellular Matrix Stiffness on Modulating the Phenotype of Macrophage

Macrophages can polarize into pro-inflammatory (M1) or anti-inflammatory (M2) phenotypes, which participate in innate immunity and play important roles in wound healing. Numerous biochemical triggers for macrophage polarization are reported, but the effects of mechanical stimuli are not fully explored yet. Concerning the rigidity alteration of microenvironment in different tissues, we investigated the M1/M2 differentiations of macrophages using rigidity-tunable engineered polymeric substrates. Macrophages showed different morphology and migration patterns, responding to the change of substrate stiffness. Quantified gene analysis indicated that cells express higher M1 phenotype markers on a softer substrate with increasing inflammatory cytokines secretion. We further induced the macrophage polarization using lipopolysaccharide (LPS) to investigate the inflammasome formation with the apoptosis-associate speck-like protein (ASC) and caspase-1, and our results showed the expressions of ASC and caspase-1 are mechanosensitive. We speculated mitochondria ROS may associate with the macrophage M1/M2 transition, and a ROS related model was proposed to interoperate the mechanosensitive of inflammasome formation. Associated with migration alteration, we also noticed the change of podosome formation and cytoskeleton assembly while cells growing on softer substrates. In conclusion, these results suggest that the macrophages are mechanosensitive, and reduced matrix stiffness can modulate macrophages to polarize to an inflammatory-like phenotype. Our findings contribute to our understanding about how matrix stiffness can differentially regulate macrophages differentiation, and are expected to provide alternative interpretations for certain disease pathogenesis.

Experimental study on the effects of mechanical stimuli on cellular UV radiation resistance of DNA

Experimental study on the effects of mechanical stimuli on cellular UV radiation resistance of DNA