Cyclic Mechanical Stress Research Articles

Biological basis of the aesthetic and rejuvenating effects in the skin after the application of the compressive microvibration Endospheres.

Compressive microvibration is a type of stimulation patented as Endospheres therapy. The stimulation, through Endospheres, applied externally to the skin surface, causes changes in the physiological and functional structure of tissues underneath the epidermis, including adipose tissue. Cyclic mechanical stress, induced by this therapy with its particular compressive microvibration, modifies the normal cell cycle of adipose tissue and promotes tissue remodelling due to the activation of the abundantly present mesenchymal-derived stem cells. In this study, a literature revision was performed to elucidate the mechanisms through which the repetitive, cyclic or modulated/ordered mechanical input of Endospheres, which results in harmonic compressive microvibrations, can have beneficial effects on the skin and all subcutaneous tissues up to the muscles, improving rejuvenation, tissue repair, vascularization and macro-organization. It was possible to formulate the hypothesis that the ordered frequency and oscillatory dynamics can completely change the chaotic and oscillatory patterns of mitochondria alongside the endoplasmic reticulum membrane in a recently discovered complex known as mitochondria-associated endoplasmatic reticulum membrane complex or mitochondria-associated endoplasmatic reticulum membrane. Analyzing the biochemical and physical characteristics of these structures, it was assessed that the nature of the benefits of the Endospheres stimulation of tissues could include three different origins: mechanical, linked to the transfer of mechanical inputs; biophysical, related to the chaotic biology of mitochondria, the generation of sinusoidal waves and the generation of piezoelectric forces; and biochemical, linked to the sensory function of adipose tissue. All these forces are discussed in detail.

Covalent Immobilization of Collagen Type I to a Polydimethylsiloxane Surface for Preventing Cell Detachment by Retaining Collagen Molecules under Uniaxial Cyclic Mechanical Stretching Stress.

Surface modification of polydimethylsiloxane (PDMS) with an extracellular matrix (ECM) is useful for enhancing stable cell attachment. However, few studies have investigated the correlation between the stability of deposited ECM and cell behavior on the PDMS surfaces in external stretched cell culture systems. Herein, covalent collagen type I (Col)-immobilized PDMS surfaces were fabricated using 3-aminopropyl-trimethoxysilane, glutaraldehyde, and Col molecules. The immobilized collagen molecules on the PDMS surface were more stable and uniform than the physisorbed collagen. The cells stably adhered to the Col-immobilized surface and proliferated even under uniaxial cyclic mechanical stretching stress (UnCyMSt), whereas the cells gradually detached from the Col-physisorbed PDMS surface, accompanied by a decrease in the number of deposited collagen molecules. Moreover, the immobilization of collagen molecules enhanced cell alignment under the UnCyMSt. This study reveals that cell adhesion, proliferation, and alignment under the UnCyMSt can be attributed to the retention of collagen molecules on the PDMS surface.

A simple sensor device for power cycle degradation sensing

A new sensor device is proposed for detection of power cycle degradation. Proposed sensor device is consisting of a Schottky-barrier MOSFET and the sensing principle is based on change of I-V characteristics by mechanical stress. Samples were fabricated by metal lift-off process and SiO2 gate insulator film deposition and the fundamental operation was evaluated by four-point bending test for mechanical stress applying. The fabricated device showed monotonically drain current change with number of stress cycles and about 5 times current change compared with initial current after 10,000 cycles of 4 N stress. Large current change by cyclic mechanical stress is attractive for a simple detection of power cycle degradation without expensive analog-circuits.

Rare protective variants and glaucoma-relevant cell stressors modulate Angiopoietin-like 7 expression.

Rare missense and nonsense variants in the Angiopoietin-like 7 (ANGPTL7) gene confer protection from primary open-angle glaucoma (POAG), though the functional mechanism remains uncharacterized. Interestingly, a larger variant effect size strongly correlates with in silico predictions of increased protein instability (r = -0.98), suggesting that protective variants lower ANGPTL7 protein levels. Here, we show that missense and nonsense variants cause aggregation of mutant ANGPTL7 protein in the endoplasmic reticulum (ER) and decreased levels of secreted protein in human trabecular meshwork (TM) cells; a lower secreted:intracellular protein ratio strongly correlates with variant effects on intraocular pressure (r = 0.81). Importantly, accumulation of mutant protein in the ER does not increase expression of ER stress proteins in TM cells (P > 0.05 for all variants tested). Cyclic mechanical stress, a glaucoma-relevant physiologic stressor, also significantly lowers ANGPTL7 expression in primary cultures of human Schlemm's canal (SC) cells (-2.4-fold-change, P = 0.01). Collectively, these data suggest that the protective effects of ANGPTL7 variants in POAG stem from lower levels of secreted protein, which may modulate responses to physiologic and pathologic ocular cell stressors. Downregulation of ANGPTL7 expression may therefore serve as a viable preventative and therapeutic strategy for this common, blinding disease.

Cyclic Stretch-Induced Mechanical Stress Applied at 1 Hz Frequency Can Alter the Metastatic Potential Properties of SAOS-2 Osteosarcoma Cells.

Recently, there has been an increasing focus on cellular morphology and mechanical behavior in order to gain a better understanding of the modulation of cell malignancy. This study used uniaxial-stretching technology to select a mechanical regimen able to elevate SAOS-2 cell migration, which is crucial in osteosarcoma cell pathology. Using confocal and atomic force microscopy, we demonstrated that a 24 h 0.5% cyclic elongation applied at 1 Hz induces morphological changes in cells. Following mechanical stimulation, the cell area enlarged, developing a more elongated shape, which disrupted the initial nuclear-to-cytoplasm ratio. The peripheral cell surface also increased its roughness. Cell-based biochemical assays and real-time PCR quantification showed that these morphologically induced changes are unrelated to the osteoblastic differentiative grade. Interestingly, two essential cell-motility properties in the modulation of the metastatic process changed following the 24 h 1 Hz mechanical stimulation. These were cell adhesion and cell migration, which, in fact, were dampened and enhanced, respectively. Notably, our results showed that the stretch-induced up-regulation of cell motility occurs through a mechanism that does not depend on matrix metalloproteinase (MMP) activity, while the inhibition of ion-stretch channels could counteract it. Overall, our results suggest that further research on mechanobiology could represent an alternative approach for the identification of novel molecular targets of osteosarcoma cell malignancy.

Recent developments in organ-on-a-chip technology for cardiovascular disease research.

Cardiovascular diseases are a group of heart and blood vessel disorders which remain a leading cause of morbidity and mortality worldwide. Currently, cardiovascular disease research commonly depends on in vivo rodent models and in vitro human cell culture models. Despite their widespread use in cardiovascular disease research, there are some long-standing limitations: animal models often fail to faithfully mimic human response, while traditional cell models ignore the in vivo microenvironment, intercellular communications, and tissue-tissue interactions. The convergence of microfabrication and tissue engineering has given rise to organ-on-a-chip technologies. The organ-on-a-chip is a microdevice containing microfluidic chips, cells, and extracellular matrix to reproduce the physiological processes of a certain part of the human body, and is nowadays considered a promising bridge between in vivo models and in vitro 2D or 3D cell culture models. Considering the difficulty in obtaining human vessel and heart samples, the development of vessel-on-a-chip and heart-on-a-chip systems can guide cardiovascular disease research in the future. In this review, we elaborate methods and materials to fabricate organ-on-a-chip systems and summarize the construction of vessel and heart chips. The construction of vessels-on-a-chip must consider the cyclic mechanical stretch and fluid shear stress, while hemodynamic forces and cardiomyocyte maturation are key factors in building hearts-on-a-chip. We also introduce the application of organs-on-a-chip in cardiovascular disease study.

Thermal fatigue and cracking behaviors of asphalt mixtures under different temperature variations

Thermal cracking is one of the most annoying distresses in asphalt pavements. The cracking of asphalt pavements caused by extremely low temperature and the damage of thermal fatigue caused by repeated temperature variations are particularly pronounced. The service condition of asphalt pavements has been affected significantly. Laboratory experiments with cyclic loads simulating the actions caused by repeated temperature variations on pavements have been conducted in many previous studies to investigate the thermal fatigue characteristics of asphalt mixtures. In those studies, an equivalent cyclic mechanical stress that can be applied by an actuator was mostly used to replace the cyclic thermal stress induced by a temperature variation. It could simplify the experimental process but could not consider the performances of asphalt mixtures varying as a function of temperature. In the present study, the thermal fatigue properties and cracking behaviors of asphalt mixtures were comprehensively investigated through a modified thermal stress restrained specimen test (TSRST) and the numerical analysis. Various temperature ranges (-20 to −10 °C, −10 to 0 °C), cyclic cooling rates (10 °C/h, 20 °C/h), low-temperature performance cooling rates (10 °C/h, 8 °C/h, 6 °C/h, 4 °C/h, 2 °C/h) and pre-crack dimensions were considered for the study to address the influences of those parameters on the analysis results. Based on the testing and simulation results, it was found that the effects of thermal fatigue on asphalt mixtures under different temperature variations and cooling rates were quantified, and the damage characteristics of the asphalt mixtures were also explored. The results showed that under the low-temperature performance state, the thermal stress of asphalt mixtures increases with a uniform cooling rate and it could be obtained that the faster the cooling rate, the larger the thermal stress generated. In the cyclic thermal stress test, due to the viscoelastic properties of asphalt mixtures, the thermal stress showed significant stress relaxation phenomena under the temperature variations. The magnitude of thermal stress decreased gradually within each cycle. It demonstrates that under the same temperature variation, the thermal stress decreases continuously with the increasing thermal cycles. Through the numerical simulation, it was clear that under different temperature ranges and cooling rates, the magnitude of thermal stress increased with the temperature range decreased, whereas the magnitude of thermal stress increased with an acceleration of the cooling rate. The lower temperature range induced a larger stress intensity factor (KI), which implies a faster cracking evolution. Analyzing the effects of different pre-crack dimensions on the number of temperature cycles, the height of pre-crack showed significant effects on the number of temperature cycles than width.

Polymerization-Induced Self-Assembly Toward Micelle-Crosslinked Tough and Ultrastretchable Hydrogels

As a promising class of tough and ultrastretchable hydrogels, micelle-crosslinked hydrogels have been restrained by the scarcity of micellar crosslinkers with a high concentration, controlled nanostructure, and uniform size distribution. Herein, polymerization-induced self-assembly (PISA) was demonstrated to be a general and powerful platform for micellar crosslinkers, affording micelle-crosslinked hydrogels with tailorable chemical structures, mechanics, and functionality. Poly(N,N-dimethylacrylamide)-b-poly(diacetone acrylamide) (PDMAc-b-PDAAM) micellar crosslinkers with a controlled nanostructure and uniform size distribution were prepared via PISA and one-step post-polymerization modification at high concentrations. Copolymerization of these micellar crosslinkers with acrylamide generated tough and ultrastretchable hydrogels, whose mechanical properties were found correlated with the concentration, nanostructure, and chemical composition of the micelles. The energy dissipation mechanism of these micelle-crosslinked hydrogels was analyzed via cyclic mechanical tests and stress relaxation experiments. The general feasibility of PISA toward micelle-crosslinked hydrogels was verified by systematic evaluation of both aqueous (including 2-methoxyethyl acrylate, tetrahydro-2-furanylmethyl acrylate, and 4-hydroxybutyl acrylate) and alcoholic (including benzyl methacrylate, lauryl methacrylate, styrene, and benzyl acrylate) PISA formulations, producing hydrogels with diverse chemical structures, mechanics, and functionalities depending on the micellar crosslinkers. The modularity of this strategy was further demonstrated by the fabrication of fluoro-functionalized hydrogels with fluoro-containing micellar crosslinkers. This strategy has significantly enlarged the scope and application of micelle-crosslinked hydrogels.

Effect of catalyst layer on fatigue life and fracture mechanisms of fuel cell membrane

AbstractPerfluorosulfonic acid (PFSA) membrane in polymer‐electrolyte fuel cells bears cyclic mechanical stress during humidity cycles under actual operation conditions, and its durability severely influences the life of fuel cells. However, the fatigue failure mechanisms of membranes are still not clear. In order to investigate the effect of catalyst layer on the fatigue life and fatigue fracture mechanisms of PFSA membranes, fatigue tests of Nafion 212 membranes and two catalyst‐coated membranes (CCMs) are conducted. The morphologies of membranes at different stages of fatigue crack initiation and propagation are identified. It is found that crack initiates from both surfaces of Nafion 212 membrane. Compared with Nafion 212 membrane, the fatigue life of single‐sided CCM increases by more than three times, while that of double‐sided CCM increases by 13 times. Such a phenomenon is ascribed to the protection of catalyst layer on membrane surface, which delays fatigue cracks initiation.

Cyclic Mechanical Strain Regulates Osteoblastic Differentiation of Mesenchymal Stem Cells on TiO2 Nanotubes Through GCN5 and Wnt/β-Catenin.

Bone marrow mesenchymal stem cells (BMSCs) play a critical role in bone formation and are extremely sensitive to external mechanical stimuli. Mechanical signals can regulate the biological behavior of cells on the surface of titanium-related prostheses and inducing osteogenic differentiation of BMSCs, which provides the integration of host bone and prosthesis benefits. But the mechanism is still unclear. In this study, BMSCs planted on the surface of TiO2 nanotubes were subjected to cyclic mechanical stress, and the related mechanisms were explored. The results of alkaline phosphatase staining, real-time PCR, and Western blot showed that cyclic mechanical stress can regulate the expression level of osteogenic differentiation markers in BMSCs on the surface of TiO2 nanotubes through Wnt/β-catenin. As an important member of the histone acetyltransferase family, GCN5 exerted regulatory effects on receiving mechanical signals. The results of the ChIP assay indicated that GCN5 could activate the Wnt promoter region. Hence, we concluded that the osteogenic differentiation ability of BMSCs on the surface of TiO2 nanotubes was enhanced under the stimulation of cyclic mechanical stress, and GCN5 mediated this process through Wnt/β-catenin.

N-acetylcysteine promotes cyclic mechanical stress-induced osteogenic differentiation of periodontal ligament stem cells by down-regulating Nrf2 expression

Background/purposeMechanical stress plays a vital role in osteogenic differentiation of periodontal ligament stem cells (PDLSCs). Cyclic mechanical stress may up-regulate reactive oxygen species (ROS) level. N-acetylcysteine (NAC) possesses powerful antioxidant capacity. However, it is undefined the impact of NAC on osteogenic differentiation stimulated by cyclic mechanical stress in PDLSCs. The aim of our research was to study the effect of NAC on PDLSCs during osteogenic differentiation under cyclic mechanical stress. Materials and methodsThe expression levels of osteogenesis markers were used to examine the osteogenic differentiation of PDLSCs. ROS production were measured by flow cytometry. The levels of reduced glutathione (GSH) and oxidized glutathione (GSSG) were analyzed. We also examined the changes of alveolar bone and periodontal ligament (PDL) tissues in orthodontic rats by micro-computed tomography (micro-CT) system and immunohistochemistry (IHC) staining. The nuclear factor erythroid-2-related factor-2 (Nrf2) expression was examined. ResultsNAC could enhance the osteogenic differentiation and up-regulate the GSH level as well as the ratio of GSH/GSSG, while down-regulate ROS generation and Nrf2 expression induced by cyclic mechanical stress in PDLSCs. NAC had beneficial effects on the microstructure of alveolar bone and enhanced the expression levels of osteogenesis markers, such as alkaline phosphatase (ALP) and collagen type 1 (COL1) in PDL in orthodontic rats at the tension side. ConclusionNAC could improve the osteogenic differentiation stimulated by cyclic mechanical stress in PDLSCs and in orthodontic rats, suggesting a potential therapeutic approach for alveolar bone remodeling in orthodontics.

Different Passive Viscoelastic Properties Between the Left and Right Ventricles in Healthy Adult Ovine.

Ventricle dysfunction is the most common cause of heart failure, which leads to high mortality and morbidity. The mechanical behavior of the ventricle is critical to its physiological function. It is known that the ventricle is anisotropic and viscoelastic. However, the understanding of ventricular viscoelasticity is much less than that of its elasticity. Moreover, the left and right ventricles (LV&RV) are different in embryologic origin, anatomy, and function, but whether they distinguish in viscoelastic properties is unclear. We hypothesized that passive viscoelasticity is different between healthy LVs and RVs. Ex vivo cyclic biaxial tensile mechanical tests (1, 0.1, 0.01 Hz) and stress relaxation (strain of 3, 6, 9, 12, 15%) were performed for ventricles from healthy adult sheep. Outflow track direction was defined as the longitudinal direction. Hysteresis stress-strain loops and stress relaxation curves were obtained to quantify the viscoelastic properties. We found that the RV had more pronounced frequency-dependent viscoelastic changes than the LV. Under the physiological frequency (1 Hz), the LV was more anisotropic in the elasticity and stiffer than the RV in both directions, whereas the RV was more anisotropic in the viscosity and more viscous than the LV in the longitudinal direction. The LV was quasi-linear viscoelastic in the longitudinal but not circumferential direction, and the RV was nonlinear viscoelastic in both directions. This study is the first to investigate passive viscoelastic differences in healthy LVs and RVs, and the findings will deepen the understanding of biomechanical mechanisms of ventricular function.

Long-Term Decrease of Intraocular Pressure in Rats by Viral Delivery of miR-146a.

PurposeTo evaluate the effects of miR-146a in trabecular meshwork (TM) cells and on intraocular pressure (IOP) in vivo via viral delivery of miR-146a to the anterior chamber of rat eyes.MethodsHuman TM cells were transfected with miR-146 mimic or inhibitor. Some cells from each group were then subjected to cyclic mechanical stress (CMS). Other cells from each group had no force applied. Gene expression was then analyzed by quantitative polymerase chain reaction (qPCR). Replication-deficient adenovirus and lentivirus expressing miR-146a were inoculated into the anterior segment of Brown Norway rat eyes. IOP was monitored by rebound tonometry, visual acuity was evaluated by optokinetic tracking (OKT), and inflammation markers in the anterior segment were examined by slit-lamp, qPCR, and semi-thin sections.ResultsmiR-146 affected the expression of genes potentially involved in outflow homeostasis at basal levels and under CMS. Both lentiviral and adenoviral vectors expressing miR-146a resulted in sustained decreases in IOP ranging from 2.6 to 4.4 mmHg. Long term follow-up of rats injected with lentiviral vectors showed a sustained effect on IOP of 4.4 ± 2.9 mmHg that lasted until rats were sacrificed more than 8 months later. Eyes showed no signs of inflammation, loss of visual acuity, or other visible abnormalities.ConclusionsIntracameral delivery of miR-146a can provide a long-term decrease of IOP in rats without signs of inflammation or other visible adverse effects.Transitional RelevanceThe IOP-lowering effects of miR-146 observed in rats provides a necessary step toward the development of an effective gene therapy for glaucoma in humans.

MiR-10396b-3p inhibits mechanical stress-induced ligamentum flavum hypertrophy by targeting IL-11.

Ligamentum flavum hypertrophy (LFH) leads to lumbar spinal stenosis (LSS) caused by LF tissue inflammation and fibrosis. Emerging evidence has indicated that dysregulated microRNAs (miRNAs) have an important role in inflammation and fibrosis. Mechanical stress (MS) has been explored as an initiating step in LFH pathology progression; the inflammation-related miRNAs induced after mechanical stress have been implicated in fibrosis pathology. However, the pathophysiological mechanism of MS-miRNAs-LFH remains to be elucidated. Using miRNAs sequencing analysis and subsequent confirmation with qRT-PCR assays, we identified the decreased expression of miR-10396b-3p and increased expression of IL-11 (interleukin-11) as responses to the development of LSS in hypertrophied LF tissues. We also found that IL-11 is positively correlated with fibrosis indicators of collagen I and collagen III. The up-regulation of miR-10396b-3p significantly decreased the level of IL-11 expression, whereas miR-10396b-3p down-regulation increased IL-11 expression in vitro. Luciferase reporter assay indicates that IL-11 is a direct target of miR-10396b-3p. Furthermore, cyclic mechanical stress inhibits miR-10396b-3p and induces IL-11, collagen I, and collagen III in vitro. Our results showed that overexpression of miR-10396b-3p suppresses MS-induced LFH by inhibiting collagen I and III via the inhibition of IL-11. These data suggest that the MS-miR-10396b-3p-IL-11 axis plays a key role in the pathological progression of LFH.

Cyclic responses and microstructure sensitivity of Cr-based turbine steel under different strain ratios in low cycle fatigue regime

When turbine steel services, i.e., startup and shutdown processes, the cyclic mechanical stress caused by the asymmetry loading conditions is introduced. To study the effect of mean strain on the cycle response and microstructure-sensitive at room temperature, the low cycle fatigue under two strain ratios was used. Experimental results indicate that significant cyclic softening was observed for both modes. The cyclic strain ratio is independent of strain amplitude and strain ratio. The plastic strain, plastic strain energy density, and cumulative strain energy density at strain ratio of 0.1 are higher than that at strain ratio of −1. Therefore, applied stress amplitude and fatigue life fitted by Manson-Coffin formula under strain ratio of 0.1 is lower than that at strain ratio of −1. According to the evaluation of the internal stress variables, i.e., back stress, isotropic stress and viscous stress under both modes, the back stress is a critical part of the cyclic stress, which can directly control the cycle softening behavior. Besides, the fatigue life prediction model considering the strain ratio is constituted through hysteresis energy method. Finally, the mechanical results are linked to the microstructure observed by transmission electron microscopy. The microstructure sensitive was evaluated and deduced.

Cyclic Mechanical Stretch Ameliorates the Degeneration of Nucleus Pulposus Cells through Promoting the ITGA2/PI3K/AKT Signaling Pathway.

Background Intervertebral disc degeneration (IVDD) is one of the major causes of low back pain and motor deficiency. Nucleus pulposus (NP) degeneration plays a key role in the process of IVDD. The mechanical and biological interactions involved in NP degeneration have not been elucidated. The present study is aimed at investigating the effect and mechanism of cyclic mechanical stretch in regulating the function and degeneration of NP cells. Methods NP cells were subjected to cyclic tensile stress (10% deformation) of 0.1 Hz for 8640 cycles. Cell proliferation was conducted through the MTT assay. The cell cycle and apoptosis were detected by flow cytometry. A gene expression profile chip was used to analyze the differentially expressed genes between the tensile stress group and the control group. Enrichment analysis of Gene Ontology (GO) annotation and signaling pathways were analyzed. Western blot and RNA interference were carried out to investigate the role of the ITGA2/PI3K/AKT pathway in the effect of cyclic mechanical stretch on NP cells. Results NP cells exhibited a greater (P < 0.05) growth rate in the tensile stress group compared to the control group. Cyclic mechanical stress significantly promoted the cell cycle transition of NP cells from the S phase to the G2/M phase. A fewer proportion of apoptotic cells were found in the tensile stress group (P < 0.05), indicating that cyclic mechanical stretch inhibits NP cell apoptosis. Microarray analysis revealed 689 significant differentially expressed genes between the two groups (P < 0.05), of which 333 genes were upregulated and another 356 genes were downregulated. Cyclic mechanical stretch altered the expression of 31 genes involved in the ITGA2/PI3K/AKT pathway and remarkably promoted this pathway in NP cells. Downregulation of ITGA2 and AKT further demonstrated that the PI3K/AKT pathway was responsible for the proliferation and COL2A1 expression of NP cells upon cyclic mechanical stretch. Conclusions Cyclic mechanical stretch promoted the proliferation and cell cycle and reversely inhibited the apoptosis of NP cells. Cyclic mechanical stretch promoted COL2A1 expression and ameliorated the degeneration of NP cells via regulation of the ITGA2/PI3K/AKT signaling pathway. Our results may provide a potential target and a possibility of IVDD disease treatment by ameliorating the degenerative changes.

Methylation-mediated down-regulation of microRNA-497-195 cluster confers osteogenic differentiation in ossification of the posterior longitudinal ligament of the spine via ADORA2A.

Aberrant expression of microRNAs (miRNAs) has been associated with spinal ossification of the posterior longitudinal ligament (OPLL). Our initial bioinformatic analysis identified differentially expressed ADORA2A in OPLL and its regulatory miRNAs miR-497 and miR-195. Hence, this study was conducted to clarify the functional relevance of miR-497-195 cluster in OPLL, which may implicate in Adenosine A2A (ADORA2A). PLL tissues were collected from OPLL and non-OPLL patients, followed by quantification of miR-497, miR-195 and ADORA2A expression. The expression of miR-497, miR-195 and/or ADORA2A was altered in posterior longitudinal ligament (PLL) cells, which then were stimulated with cyclic mechanical stress (CMS). We validated that ADORA2A was expressed highly, while miR-497 and miR-195 were down-regulated in PLL tissues of OPLL patients. miR-195 and miR-497 expression in CMS-treated PLL cells was restored by a demethylation reagent 5-aza-2'-deoxycytidine (AZA). Moreover, expression of miR-195 and miR-497 was decreased by promoting promoter CpG island methylation. ADORA2A was verified as the target of miR-195 and miR-497. Overexpression of miR-195 and miR-497 diminished expression of osteogenic factors in PLL cells by inactivating the cAMP/PKA signaling pathway via down-regulation of ADORA2A. Collectively, miR-497-195 cluster augments osteogenic differentiation of PLL cells by inhibiting ADORA2A-dependent cAMP/PKA signaling pathway.

SB772077B (SB77) Alleviated the Aqueous Outflow Resistance Mediated by Cyclic Mechanical Stress in Perfused Human Cadaveric Eyes

The intraocular pressure lowering property of a new rho kinase inhibitor, SB772077B (SB77) has been previously demonstrated in perfused human cadaveric eyes. In this study, the efficacy of SB77 in alleviating the aqueous outflow resistance mediated by cyclic mechanical stress in perfused human cadaveric eyes was investigated. A human anterior segment perfusion culture model was used to investigate the effect of cyclic intraocular pressure (IOP) on aqueous outflow facility in presence or absence of SB77. The status of RhoA activation and the downstream effector molecule myosin-light chain phosphorylation (p-MLC) was investigated by Western blot. Cyclic mechanical stress resulted in decrease in aqueous outflow facility (–19.79 ± 4.93%; p = 0.019) in perfused human eyes and treatment with SB77 (50 µM) significantly enhanced outflow facility by 15% (p = 0.05). The increase in outflow facility by SB77 was confirmed with the inactivation of RhoA/ROCK signaling and decreased expression of extracellular matrix markers. SB77 effectively reduced the outflow resistance mediated by cyclic IOP and thus may be a potential clinical candidate for the management of glaucoma.

The long noncoding RNA H19 attenuates force-driven cartilage degeneration via miR-483–5p/Dusp5

Developmental dysplasia of the hip (DDH) is a common hip disease characterized by abnormal development of the acetabulum and femoral head. In most cases, DDH ultimately leads to osteoarthritis. Anomalous biomechanical force plays an important role in cartilage degeneration in DDH. However, in addition to mechanical wear, the underlying molecular mechanisms in cartilage degeneration in DDH remain unclear. This study analyzed the effect of long noncoding RNA (lncRNA)-H19 on DDH cartilage degradation. To elucidate the specific role of lncRNA H19, we established an intermittent cyclic mechanical stress (ICMS) cell force model to simulate abnormal biomechanical environment in vitro. Then, the roles of lncRNA-H19 were also determined in vivo by establishing a model of swaddling DDH. We observed that patients with DDH possessed low levels of lncRNA-H19, COL2A1, and Aggrecan but high levels of MMP3 and Adamts5. The same results were also obtained in a DDH rat model. Furthermore, the data suggested that ICMS promoted cartilage degeneration and caused reorientation of the cytoskeleton, and lncRNA H19 helped inhibit cartilage degeneration. Bioinformatics analysis and lncRNA sequencing were performed, and luciferase assays showed that lncRNA H19 and Dusp5 are both direct targets of miR-483–5p. Moreover, Dups5 plays a negative role in ICMS-induced cartilage degradation by activating the Erk and p38 pathways. In vivo, lncRNA H19 had protective effects on the swaddling DDH model. These findings indicate that lncRNA-H19 played a positive role in cartilage degradation in DDH through the lncRNA H19/miR-483–5p/Dusp5 axis.

Mechanical loading induces HIF-1α expression in chondrocytes via YAP.

To investigate the role of YAP in cyclic mechanical stress induced up-regulation of HIF-1α in rat cartilage chondrocytes. Our in vitro and in vivo experiments demonstrated that cyclic mechanical stress promoted HIF-1α and YAP proteins expression in a magnitude dependent manner. Cyclic mechanical stress at 4000μ strain exhibited most significant effect in promoting HIF-1α and YAP up-regulation. Activation of YAP using LPA significantly promoted HIF-1α stabilization and expression, while YAP siRNA treatment suppressed the up-regulation of HIF-1α induced by cyclic mechanical stress. Our results indicated that cyclic mechanical stress promoted HIF-1α stabilization and YAP is involved in mechanical stress induced HIF-1α up-regulation.