Effect Of Fluid Shear Stress Research Articles

Effects of fluid shear stress on oral biofilm formation and composition and the transcriptional response of Streptococcus gordonii.

Biofilms are subjected to many environmental pressures that can influence community structure and physiology. In the oral cavity, and many other environments, biofilms are exposed to forces generated by fluid flow; however, our understanding of how oral biofilms respond to these forces remains limited. In this study, we developed a linear rocker model of fluid flow to study the impact of shear forces on Streptococcus gordonii and dental plaque-derived multispecies biofilms. We observed that as shear forces increased, S. gordonii biofilm biomass decreased. Reduced biomass was largely independent of overall bacterial growth. Transcriptome analysis of S. gordonii biofilms exposed to moderate levels of shear stress uncovered numerous genes with differential expression under shear. We also evaluated an ex vivo plaque biofilm exposed to fluid shear forces. Like S. gordonii, the plaque biofilm displayed decreased biomass as shear forces increased. Examination of plaque community composition revealed decreased diversity and compositional changes in the plaque biofilm exposed to shear. These studies help to elucidate the impact of fluid shear on oral bacteria and may be extended to other bacterial biofilm systems.

Overcoming the effects of fluid shear stress in ovarian cancer cell lines: Doxorubicin alone or photodynamic priming to target platinum resistance

AbstractResistance to platinum‐based chemotherapies remains a significant challenge in advanced‐stage high‐grade serous ovarian carcinoma, and patients with malignant ascites face the poorest outcomes. It is, therefore, important to understand the effects of ascites, including the associated fluid shear stress (FSS), on phenotypic changes and therapy response, specifically FSS‐induced chemotherapy resistance and the underlying mechanisms in ovarian cancer. This study investigated the effects of FSS on response to cisplatin, a platinum‐based chemotherapy, and doxorubicin, an anthracycline, both of which are commonly used to manage advanced‐stage ovarian cancer. Consistent with prior research, OVCAR‐3 and Caov‐3 cells cultivated under FSS demonstrated significant resistance to cisplatin. Examination of the role of mitochondria revealed an increase in mitochondrial DNA copy number and intracellular ATP content in cultures grown under FSS, suggesting that changes in mitochondria number and metabolic activity may contribute to platinum resistance. Interestingly, no resistance to doxorubicin was observed under FSS, the first such observation of a lack of resistance under these conditions. Finally, this study demonstrated the potential of photodynamic priming using benzoporphyrin derivative, a clinically approved photosensitizer that localizes in part to mitochondria and endoplasmic reticula, to enhance the efficacy of cisplatin, but not doxorubicin, thereby overcoming FSS‐induced platinum resistance.

Retracted: Effects of Fluid Shear Stress on Human Intervertebral Disc Nucleus Pulposus Cells Based on Label-Free Quantitative Proteomics.

[This retracts the article DOI: 10.1155/2022/3860898.].

Notch signaling and fluid shear stress in regulating osteogenic differentiation.

Osteoporosis is a common bone and metabolic disease that is characterized by bone density loss and microstructural degeneration. Human bone marrow-derived mesenchymal stem cells (hMSCs) are multipotent progenitor cells with the potential to differentiate into various cell types, including osteoblasts, chondrocytes, and adipocytes, which have been utilized extensively in the field of bone tissue engineering and cell-based therapy. Although fluid shear stress plays an important role in bone osteogenic differentiation, the cellular and molecular mechanisms underlying this effect remain poorly understood. Here, a locked nucleic acid (LNA)/DNA nanobiosensor was exploited to monitor mRNA gene expression of hMSCs that were exposed to physiologically relevant fluid shear stress to examine the regulatory role of Notch signaling during osteogenic differentiation. First, the effects of fluid shear stress on cell viability, proliferation, morphology, and osteogenic differentiation were investigated and compared. Our results showed shear stress modulates hMSCs morphology and osteogenic differentiation depending on the applied shear and duration. By incorporating this LNA/DNA nanobiosensor and alkaline phosphatase (ALP) staining, we further investigated the role of Notch signaling in regulating osteogenic differentiation. Pharmacological treatment is applied to disrupt Notch signaling to investigate the mechanisms that govern shear stress induced osteogenic differentiation. Our experimental results provide convincing evidence supporting that physiologically relevant shear stress regulates osteogenic differentiation through Notch signaling. Inhibition of Notch signaling mediates the effects of shear stress on osteogenic differentiation, with reduced ALP enzyme activity and decreased Dll4 mRNA expression. In conclusion, our results will add new information concerning osteogenic differentiation of hMSCs under shear stress and the regulatory role of Notch signaling. Further studies may elucidate the mechanisms underlying the mechanosensitive role of Notch signaling in stem cell differentiation.

A novel ceRNA regulatory network involving the long noncoding NEAT1, miRNA-466f-3p and its mRNA target in osteoblast autophagy and osteoporosis.

Osteoporosis (OP) is a systemic metabolic disorder characterized by a reduction in bone tissue volume. LncRNAs have been reported to act as regulators of several human diseases. Specifically, lncRNA nuclear paraspeckle assembly transcript 1 (NEAT1) is involved in proliferation, differentiation and apoptosis in osteoclasts and bone marrow mesenchymal stem cells and regulates the occurrence and development of OP. However, the relationship between NEAT1 and osteoblast autophagy and its mechanism are still unclear. Western blotting of LC3 and P62 was used to evaluate the effect of fluid shear stress (FSS) on autophagy in MC3T3-E1 osteoblasts. Total transcriptome sequencing and bioinformatics analyses were performed on osteoblasts loaded with and without FSS. qPCR was performed to examine the expression of NEAT1 in OP bone tissues and osteoblasts. The RNA-FISH was performed to study the localization of lncRNA NEAT1 and miR-466f-3p in MC3T3-E1 osteoblasts. In vitro, western blotting, transmission electron microscopy (TEM), immunofluorescence (IF) staining and qPCR were performed to verify the biological functions of NEAT1, miR-466f-3p and HK2. Subsequently, we conducted bioinformatics analysis and dual luciferase reporter assays to identify the relationships among NEAT1, miR-466f-3p and HK2. Additionally, rescue assays were conducted on osteoblasts to clarify the regulatory network of the NEAT1/miR-466f-3p/HK2 signalling pathway. In vivo, the OVX mouse model was used to investigate the effects of si-NEAT1 on autophagy in OP mice. The distal femur and serum were collected for further micro-CT analysis, blood biochemistry, and haematoxylin-eosin and Alizarin red staining (ARS). Immunohistochemistry (IHC) was performed to assess the protein expression of LC3 and HK2. NEAT1 expression was upregulated in OP tissues and osteoblast lines exposed to FSS. Knockdown of NEAT1 inhibited autophagy in vitro and in vivo. Further studies demonstrated that NEAT1 positively regulated HK2 expression via its competing endogenous RNA effects on miR-466f-3p. Moreover, we found the NEAT1/miR-466f-3p/HK2 axis regulated autophagy in osteoblasts. Knocking down NEAT1 inhibited autophagy in osteoblasts via the miR-466f-3p/HK2 signalling pathway, which may provide new ideas for novel molecular therapeutic targets of postmenopausal OP. KEY MESSAGES: • Fluid shear stress (FSS) can promote autophagy of osteoblast and performed transcriptome sequencing. • NEAT1 is overexpressed in osteoporosis and could regulate osteoblast cells autophagy. • Knockdown of lncRNA NEAT1 inhibited osteoblast cells autophagy by sponging miRNA-466f-3p and targeting HK2 in osteoporosis.

Effects of Fluid Shear Stress on Human Intervertebral Disc Nucleus Pulposus Cells Based on Label-Free Quantitative Proteomics.

Objective To explore the possible mechanism of fluid shear stress on human nucleus pulposus cells based on label-free proteomics technology. Methods The human nucleus pulposus cell line was purchased and subcultured in vitro. The Flexcell STR-4000 multiflow field cell fluid shear stress loading culture system was used to apply continuous laminar fluid shear stress (12 dyne/cm2, 45 mins) to the monolayer adherent cells. Those without mechanical loading were used as the control group, and those subjected to fluid shear loading were used as the experimental group. Differential protein expression was identified using mass spectrometry identification technology, and bioinformatics analysis was performed using Gene Ontology GO (Gene Ontology) and Kyoto Encyclopedia of Genes and Genomes KEGG (Kyoto Encyclopedia of Genes and Genomes). Results The proteomics results of the experimental group and the control group showed that the total number of mass spectra was 638653, the number of matched mass spectra was 170110, the total number of identified peptides was 32050, the specific peptide was 30564, and the total number of identified proteins was 4745. Comparing the two groups, 47 proteins were significantly differentially expressed, namely, 25 upregulated proteins and 22 downregulated proteins. Bioinformatics analysis showed that significantly different proteins were mainly manifested in cellular process, biological regulation, metabolic process, binding, catalytic activity, cellular components (cell part), organelle part (organelle part), and other molecular biological functions. Conclusion Using proteomics technology to screen human nucleus pulposus cells after fluid shear stress loading, the differential protein expression provides a basis for further exploration of the mechanism of mechanical factors on nucleus pulposus.

Synergistic effects of fluid shear stress and adhesion morphology on the apoptosis and osteogenesis of mesenchymal stem cells.

Mechanical microenvironments, such as characteristics defining mechanical environments and fluid flow play an important role in steering the fate of mesenchymal stem cells (MSCs). However, the synergistic effect of adhesion morphology and fluid flow on the biological behavior of MSCs is seldom investigated. In this article, 0.5 or 0.8Pa fluid shear stress (FSS) was applied to the MSCs on micropatterned substrates, and the apoptosis and osteogenic differentiation of MSCs were measured by double fluorescent staining. Results showed that the cellular adhesion patterns with low circularity and large area are beneficial to the osteogenic differentiation of individual MSCs. Meanwhile, FSS facilitated osteogenic differentiation of MSCs, as shown by the expression of alkaline phosphatase, osteocalcin, and collagen I. In addition, nuclear transfer of Yes-associated protein, a transcriptional regulator in MSCs, was enhanced after being exposed to FSS. These results demonstrated the synergistic effects of FSS and adhesion morphology in directing the fate of MSCs, and these effects may be adopted to design bio-functional substrates for cell transplantation in tissue engineering.

Effect of Fluid Shear Stress on Human Breast Cancer Cells

The importance of Hemodynamic forces such as interstitial and vascular Fluid Shear Stress (FSS), which are exposed to cancer cells, is not well understood. Here, we report that human breast cancer cells, MDA‐MB‐231, show increased cytosolic and nuclear area, nuclear flattening, focal adhesion and stress fiber formation in the presence of 6 dyn/cm2 orbital shear stress. Withdrawal of FSS restores the morphology within 12 hours. Time lapse video microscopy reveals that FSS pre‐exposed cells migrate more slowly compared with the control cells which are not pre‐exposed with FSS. Interestingly, when we seed these cells on soft agar plate, cells are able to grow, and the size of the colonies are significantly larger compared with the control cells. The transcriptional co‐activator, responsive to biomechanical cues, YAP‐1 translocates into nucleus in the presence of FSS. Altogether, these data suggest that FSS can change homeostasis of cancer cells to facilitate their survival.

Revealing anisotropic elasticity of endothelium under fluid shear stress

Endothelium lining interior surface of blood vessels experiences various physical stimulations in vivo. Its physical properties, especially elasticity, play important roles in regulating the physiological functions of vascular systems. In this paper, an integrated approach is developed to characterize the anisotropic elasticity of the endothelium under physiological-level fluid shear stress. A pressure sensor-embedded microfluidic device is developed to provide fluid shear stress on the perfusion-cultured endothelium and to measure transverse in-plane elasticities in the directions parallel and perpendicular to the flow direction. Biological atomic force microscopy (Bio-AFM) is further exploited to measure the vertical elasticity of the endothelium in its out-of-plane direction. The results show that the transverse elasticity of the endothelium in the direction parallel to the perfusion culture flow direction is about 70% higher than that in the direction perpendicular to the flow direction. Moreover, the transverse elasticities of the endothelium are estimated to be approximately 120 times larger than the vertical one. The results indicate the effects of fluid shear stress on the transverse elasticity anisotropy of the endothelium, and the difference between the elasticities in transverse and vertical directions. The quantitative measurement of the endothelium anisotropic elasticity in different directions at the tissue level under the fluid shear stress provides biologists insightful information for the advanced vascular system studies from biophysical and biomaterial viewpoints. Statement of SignificanceIn this paper, we take advantage an integrated approach combining microfluidic devices and biological atomic force microscopy (Bio-AFM) to characterize anisotropic elasticities of endothelia with and without fluidic shear stress application. The microfluidic devices are exploited to conduct perfusion cell culture of the endothelial cells, and to estimate the in-plane elasticities of the endothelium in the direction parallel and perpendicular to the shear stress. In addition, the Bio-AFM is utilized for characterization of the endothelium morphology and vertical elasticity. The measurement results demonstrate the very first anisotropic elasticity quantification of the endothelia. Furthermore, the study provides insightful information bridging the microscopic sing cell and macroscopic organ level studies, which can greatly help to advance vascular system research from material perspective.

Effects of Fluid Shear Stress on Vascular Endothelial Cells

Effects of Fluid Shear Stress on Vascular Endothelial Cells

Effects of Stress on Osteoblast Proliferation and Differentiation Based on Endoplasmic Reticulum Stress and Wntβ-Catenin Signaling Pathway.

In order to investigate the effect of fluid shear stress on the proliferation of osteoblasts and the regulatory role of the Wnt/β-catenin signaling pathway in cell proliferation, a new method based on endoplasmic reticulum stress and Wnt/β-catenin signaling pathway stress-mediated was proposed. Taking MG63 osteoblasts as the research object, they were inoculated on glass slides (G group), polished titanium sheets (P group), and sandblasted acid-base treated pure titanium sheets (S group). In addition, FSS of 0 dunes/cm2 (static group) and 12 dunes/cm2 (stress group) were given, respectively. Then, quantitative reverse transcription-PCR (RT-qPCR) and western blot were used to detect the mRNA and protein expressions of low-density lipoprotein receptor-related protein 5 (LRP5) and β-catenin in MG63 cells. The results showed that the expression levels of β-catenin mRNA and protein in cells in the stress group were significantly increased (P < 0.05), and the protein expression level of LRP5 was significantly decreased (P < 0.05). The expression level of LRP5 in group S was greatly inhibited, while the expression level of β-catenin was significantly upregulated. Therefore, FSS can stimulate the expression of LRP5 and β-catenin in osteoblasts. Fluid shear stress can promote osteoblast proliferation in vitro; the Wnt/β-catenin signaling pathway is involved in regulating fluid shear stress to promote osteoblast proliferation.

Fluid shear stress and endothelial cells synergistically promote osteogenesis of mesenchymal stem cells via integrin β1-FAK-ERK1/2 pathway.

Prevascularization and mechanical stimulation have been reported as effective methods for the construction of functional bone tissue. However, their combined effects on osteogenic differentiation and its mechanism remain to be explored. Here, the effects of fluid shear stress (FSS) on osteogenic differentiation of rat bone-marrow-derived mesenchymal stem cells (BMSCs) when cocultured with human umbilical vein endothelial cells (HUVECs) were investigated, and underlying signaling mechanisms were further explored. FSS stimulation for 1–4 h/day increased alkaline phosphatase (ALP) activity and calcium deposition in coculture systems and promoted the proliferation of cocultured cells. FSS stimulation for 2 h/day was selected as the optimized protocol according to osteogenesis in the coculture. In this situation, the mRNA levels of ALP, runt-related transcriptional factor 2 (Runx2) and osteocalcin (OCN), and protein levels of OCN and osteopontin (OPN) in BMSCs were upregulated. Furthermore, FSS and coculture with HUVECs synergistically increased integrin β1 expression in BMSCs and further activated focal adhesion kinases (FAKs) and downstream extracellular signal-related kinase (ERK), leading to the enhancement of Runx2 expression. Blocking the phosphorylation of FAK abrogated FSS-induced ERK phosphorylation and inhibited osteogenesis of cocultured BMSCs. These results revealed that FSS and coculture with HUVECs synergistically promotes the osteogenesis of BMSCs, which was mediated by the integrin β1-FAK-ERK signaling pathway.

The Response of Corneal Endothelial Cells to Shear Stress in an In Vitro Flow Model.

Purpose Corneal endothelial cells are usually exposed to shear stress caused by the aqueous humour, which is similar to the exposure of vascular endothelial cells to shear stress caused by blood flow. However, the effect of fluid shear stress on corneal endothelial cells is still poorly understood. The purpose of this study was to explore whether the shear stress that results from the aqueous humour influences corneal endothelial cells. Methods An in vitro model was established to generate fluid flow on cells, and the effect of fluid flow on corneal endothelial cells after exposure to two levels of shear stress for different durations was investigated. The mRNA and protein expression of corneal endothelium-related markers in rabbit corneal endothelial cells was evaluated by real-time PCR and western blotting. Results The expression of the corneal endothelium-related markers ZO-1, N-cadherin, and Na+-K+-ATPase in rabbit corneal endothelial cells (RCECs) was upregulated at both the mRNA and protein levels after exposure to shear stress. Conclusion This study demonstrates that RCECs respond favourably to fluid shear stress, which may contribute to the maintenance of corneal endothelial cell function. Furthermore, this study also provides a theoretical foundation for further investigating the response of human corneal endothelial cells to the shear stress caused by the aqueous humour.

Mechanical and chemical cues synergistically promote human venous smooth muscle cell osteogenesis through integrin β1-ERK1/2 signaling: A cell model of hemodialysis fistula calcification.

Arteriovenous fistula (AVF) is the vascular access of choice for renal replacement therapy. However, AVF is susceptible to calcification with a high prevalence of 40%-65% in chronic hemodialysis patients. Repeated needle puncture for hemodialysis cannulation results in intimal denudation of AVF. We hypothesized that exposure to blood shear stress in the medial layer promotes venous smooth muscle cell (SMC) osteogenesis. While previous studies of shear stress focused on arterial-type SMCs, SMCs isolated from the vein had not been investigated. This study established a venous cell model of AVF using the fluid shear device, combined with a high phosphate medium to mimic the uremic milieu. Osteogenic gene expression of venous SMCs upon mechanical and chemical cues was analyzed in addition to the activated cell signaling pathways. Our findings indicated that upon shear stress and high phosphate environment, mechanical stimulation (shear stress) had an additive effect in up-regulation of an early osteogenic marker, Runx2. We further identified that the integrin β1-ERK1/2signaling pathway was responsible for the molecular basis of venous SMC osteogenesis upon shear stress exposure. Mitochondrial biogenesis also took part in the early stage of this venopathy pathogenesis, evident by the up-regulated mitochondrial transcription factor A and mitochondrial DNA polymerase γ in venous SMCs. In conclusion, synergistic effects of fluid shear stress and high phosphate induce venous SMC osteogenesis via the ERK1/2 pathway through activating the mechanosensing integrin β1signaling. The present study identified a promising druggable target for reducing AVF calcification, which deserves further in vivo investigations.

Cell spinpods are a simple inexpensive suspension culture device to deliver fluid shear stress to renal proximal tubular cells

Rotating forms of suspension culture allow cells to aggregate into spheroids, prevent the de-differentiating influence of 2D culture, and, perhaps most importantly of all, provide physiologically relevant, in vivo levels of shear stress. Rotating suspension culture technology has not been widely implemented, in large part because the vessels are prohibitively expensive, labor-intensive to use, and are difficult to scale for industrial applications. Our solution addresses each of these challenges in a new vessel called a cell spinpod. These small 3.5 mL capacity vessels are constructed from injection-molded thermoplastic polymer components. They contain self-sealing axial silicone rubber ports, and fluoropolymer, breathable membranes. Here we report the two-fluid modeling of the flow and stresses in cell spinpods. Cell spinpods were used to demonstrate the effect of fluid shear stress on renal cell gene expression and cellular functions, particularly membrane and xenobiotic transporters, mitochondrial function, and myeloma light chain, cisplatin and doxorubicin, toxicity. During exposure to myeloma immunoglobulin light chains, rotation increased release of clinically validated nephrotoxicity cytokine markers in a toxin-specific pattern. Addition of cisplatin or doxorubicin nephrotoxins reversed the enhanced glucose and albumin uptake induced by fluid shear stress in rotating cell spinpod cultures. Cell spinpods are a simple, inexpensive, easily automated culture device that enhances cellular functions for in vitro studies of nephrotoxicity.

Numerical Investigation of Wave-Induced Uplift Force Exerted onto a Shallowly Buried Pipeline Using a Two-Dimensional Fully Coupled Model for Wave-Seabed-Structure Interaction

Duan, L.; Xing, L.; Wang, D.; Fan, M., and Wang, P., 2021. Numerical investigation of wave-induced uplift force exerted onto a shallowly buried pipeline using a two-dimensional fully coupled model for wave-seabed-structure interaction. Journal of Coastal Research, 37(4), 800–812. Coconut Creek (Florida), ISSN 0749-0208.In this paper, a novel two-dimensional numerical model for wave-seabed-pipeline interaction is proposed to investigate the wave-induced uplift force exerted onto a shallowly buried pipeline. Unlike previous models, the present model is fully coupled, and the effects of fluid shear stress at the seabed surface on the soil dynamic response are considered. In this study, the Reynolds-averaged Navier-Stokes equation was applied to simulate flow motions, and the level set method was used to track the free water surface. Meanwhile, Biot's poroelastic equation (u-p mode) was taken to govern the wave-induced oscillatory response of the seabed, and the pipeline was treated as elastic. After validation using data from laboratory experiments in the literature, the novel model was first employed to investigate the distribution of the wave-induced transient seabed response around the shallowly buried pipeline. Then, the effects of the wave parameters, the soil properties, and the pipeline diameter and burial depth on the wave-induced maximum uplift force exerted onto the structure were systematically examined in detail. Finally, wave-induced maximum uplift of the shallowly buried pipeline was predicted based on the numerical results, which may provide some references for engineering practice. Numerical results confirmed that both the nonlinear pipeline-seabed contacts and the fluid shear stress at the seabed surface can greatly affect the soil dynamic response, which should be considered in determining the wave-induced uplift force exerted onto a shallowly buried pipeline.

Effects of fluid shear stress on expression of focal adhesion kinase in MG-63 human osteoblast-like cells on different surface modification of titanium

ABSTRACT This study aimed to investigate the effect of fluid shear stress (FSS) on cell proliferation and expression of focal adhesion kinase (FAK) in MG-63 cells on different modified titanium surfaces. MG63 cells were cultured on three different surfaces: glass slide, polished treatment (PT) titanium surface and sandblasted/acid-etched surfaces (SLA) titanium surface. The surface topography and roughness were evaluated by scanning electron microscopy (SEM) and atomic force microscopy (AFM), respectively. The cells were subjected to FSS, and the cell appearance before and after the stress was evaluated. MTT assay was applied to estimate cell proliferation. The mRNA and protein levels of FAK were determined by qRT-PCR and western blotting. Titanium plates demonstrated different surface microtopography. Parameter Ra values of SLA group were around 3.4 µm, which was higher than PT group. Exposure to the FSS of 12 dynes/cm2 significantly induced positive upregulation of cellular proliferation and the expression of FAK, which were directly correlated with the duration of exposure and surface. Cells in SLA group were able to endurance the longtime of FSS, especially under the FSS of 16 dynes/cm2. SLA surface had a positive influence on the expression of FAK. Different surface modifications created different microtopography of titanium plates. Cell proliferation and the mRNA and protein expression of FAK were stimulated by FSS and regulated by a marked synergistic effect of surface topography and the level and duration of FSS.

Shear Stress Modulates Osteoblast Cell and Nucleus Morphology and Volume.

Mechanical loading preserves bone mass and function—yet, little is known about the cell biological basis behind this preservation. For example, cell and nucleus morphology are critically important for cell function, but how these morphological characteristics are affected by the physiological mechanical loading of bone cells is under-investigated. This study aims to determine the effects of fluid shear stress on cell and nucleus morphology and volume of osteoblasts, and how these effects relate to changes in actin cytoskeleton and focal adhesion formation. Mouse calvaria 3T3-E1 (MC3T3-E1) osteoblasts were treated with or without 1 h pulsating fluid flow (PFF). Live-cell imaging was performed every 10 min during PFF and immediately after PFF. Cytoskeletal organization and focal adhesions were visualized, and gene and protein expression quantified. Two-dimensional (2D) and three-dimensional (3D) morphometric analyses were made using MeasureStack and medical imaging interaction toolkit (MITK) software. 2D-images revealed that 1 h PFF changed cell morphology from polygonal to triangular, and nucleus morphology from round to ellipsoid. PFF also reduced cell surface area (0.3-fold), cell volume (0.3-fold), and nucleus volume (0.2-fold). During PFF, the live-cell volume gradually decreased from 6000 to 3000 µm3. After PFF, α-tubulin orientation was more disorganized, but F-actin fluorescence intensity was enhanced, particularly around the nucleus. 3D-images obtained from Z-stacks indicated that PFF increased F-actin fluorescence signal distribution around the nucleus in the XZ and YZ direction (2.3-fold). PFF increased protein expression of phospho-paxillin (2.0-fold) and integrin-α5 (2.8-fold), but did not increase mRNA expression of paxillin-a (PXNA), paxillin-b (PXNB), integrin-α5 (ITGA51), or α-tubulin protein expression. In conclusion, PFF induced substantial changes in osteoblast cytoskeleton, as well as cell and nucleus morphology and volume, which was accompanied by elevated gene and protein expression of adhesion and structural proteins. More insights into the mechanisms whereby mechanical cues drive morphological changes in bone cells, and thereby, possibly in bone cell behavior, will aid the guidance of clinical treatment, particularly in the field of orthodontics, (oral) implantology, and orthopedics.

The effect of fluid shear stress in hydrogen sulphide production and cystathionine γ-lyase expression in human early endothelial progenitor cells.

BackgroundPhysiological fluid shear stress has been shown to have a beneficial impact on vascular homeostasis. Endothelial progenitor cells (EPCs) make a significant contribution to maintaining endothelial integrity. Therefore, we hypothesised that shear stress-induced endothelium protection plays a role in hydrogen sulphide (H2S) production and up-regulation of cystathionine γ-lyase (CSE) expression in EPCs.MethodsHuman EPC-derived CSE activity was detected by colorimetric assay, and H2S production was evaluated by membrane adsorption method. Cell proliferation, migration, and adhesion were assessed by MTT, Transwell, and endothelial cell-mediated adhesion assays, respectively. Real-time polymerase chain reaction (RT-PCR) was carried out to analyse gene expression. Protein expression was analysed by western blot.ResultsHuman EPCs were treated with shear stress levels of 5–25 dyn/cm2 for up to 3 h, and 25 dyn/cm2 for up to 24 h. H2S production and CSE mRNA expression in the EPCs were increased by shear stress in a dose-dependent manner in vitro. Likewise, time-dependent shear stress also significantly enhanced CSE protein expression. Compared to static condition, shear stress improved EPCs proliferation, migration and adhesion capacity. Knockdown of CSE expression by small interfering RNA substantially eliminated the shear stress-induced above functions of human EPCs in vitro.ConclusionsThis study gives new insight into the regulatory effect of physiological shear stress on the CSE/H2S system in human EPCs. Our findings may contribute to the development of vascular protective research, although the relevant evidence is admittedly indirect.

Flow-induced Shear Stress Confers Resistance to Carboplatin in an Adherent Three-Dimensional Model for Ovarian Cancer: A Role for EGFR-Targeted Photoimmunotherapy Informed by Physical Stress.

A key reason for the persistently grim statistics associated with metastatic ovarian cancer is resistance to conventional agents, including platinum-based chemotherapies. A major source of treatment failure is the high degree of genetic and molecular heterogeneity, which results from significant underlying genomic instability, as well as stromal and physical cues in the microenvironment. Ovarian cancer commonly disseminates via transcoelomic routes to distant sites, which is associated with the frequent production of malignant ascites, as well as the poorest prognosis. In addition to providing a cell and protein-rich environment for cancer growth and progression, ascitic fluid also confers physical stress on tumors. An understudied area in ovarian cancer research is the impact of fluid shear stress on treatment failure. Here, we investigate the effect of fluid shear stress on response to platinum-based chemotherapy and the modulation of molecular pathways associated with aggressive disease in a perfusion model for adherent 3D ovarian cancer nodules. Resistance to carboplatin is observed under flow with a concomitant increase in the expression and activation of the epidermal growth factor receptor (EGFR) as well as downstream signaling members mitogen-activated protein kinase/extracellular signal-regulated kinase (MEK) and extracellular signal-regulated kinase (ERK). The uptake of platinum by the 3D ovarian cancer nodules was significantly higher in flow cultures compared to static cultures. A downregulation of phospho-focal adhesion kinase (p-FAK), vinculin, and phospho-paxillin was observed following carboplatin treatment in both flow and static cultures. Interestingly, low-dose anti-EGFR photoimmunotherapy (PIT), a targeted photochemical modality, was found to be equally effective in ovarian tumors grown under flow and static conditions. These findings highlight the need to further develop PIT-based combinations that target the EGFR, and sensitize ovarian cancers to chemotherapy in the context of flow-induced shear stress.