Microgravity and bone cell mechanosensitivity: FLOW experiment during the DELTA mission

The catabolic effects of microgravity on mineral metabolism in bone organ cultures might be explained as resulting from an exceptional form of disuse. It is possible that the mechanosensitivity of bone cells is altered under near weightlessness conditions, which likely contributes to disturbed bone metabolism observed in astronauts. In the experiment “FLOW”, we tested whether the production of early signaling molecules that are involved in the mechanical load-induced osteogenic response by bone cells is changed under microgravity conditions. FLOW was one of the Biological experiment entries to the Dutch Soyuz Mission “DELTA” (Dutch Expedition for Life Science, Technology and Atmospheric Research). FLOW was flown by the Soyuz craft, launched on April 19, 2004, on its way to the International Space Station. Primary osteocytes, osteoblasts, and periosteal fibroblasts were incubated in plunger boxes, developed by Centre for Concepts in Mechatronics, using plunger activation events for single pulse fluid shear stress stimulations. Due to unforeseen hardware complications, results from in-flight cultures are considered lost. Ground control experiments showed an accumulative increase of NO in medium for osteocytes (as well as for osteoblasts and periosteal fibroblasts). Data from the online-NO sensor showed that the NO produced in medium by osteocytes increased sharply after pulse shear stress stimulations. COX-2 mRNA expression revealed high levels in osteoblasts compared to the other cell types tested. In conclusion, preparations for the FLOW experiment and preliminary ground results indicate that the FLOW setup is viable for a future flight opportunity.     

Shear-induced force transmission in a multicomponent,multicell model of the endothelium

Haemodynamic forces applied at the apical surface of vascular endothelial cells (ECs) provide the mechanical signals at intracellular organelles and through the inter-connected cellular network. The objective of this study is to quantify the intracellular and intercellular stresses in a confluent vascular EC monolayer. A novel three-dimensional, multiscale and multicomponent model of focally adhered ECs is developed to account for the role of potential mechanosensors (glycocalyx layer, actin cortical layer, nucleus, cytoskeleton, focal adhesions (FAs) and adherens junctions (ADJs)) in mechanotransmission and EC deformation. The overriding issue addressed is the stress amplification in these regions, which may play a role in subcellular localization of mechanotransmission. The model predicts that the stresses are amplified 250–600-fold over apical values at ADJs and 175–200-fold at FAs for ECs exposed to a mean shear stress of 10 dyne cm⁻². Estimates of forces per molecule in the cell attachment points to the external cellular matrix and cell–cell adhesion points are of the order of 8 pN at FAs and as high as 3 pN at ADJs, suggesting that direct force-induced mechanotransmission by single molecules is possible in both. The maximum deformation of an EC in the monolayer is calculated as 400 nm in response to a mean shear stress of 1 Pa applied over the EC surface which is in accord with measurements. The model also predicts that the magnitude of the cell–cell junction inclination angle is independent of the cytoskeleton and glycocalyx. The inclination angle of the cell–cell junction is calculated to be 6.6° in an EC monolayer, which is somewhat below the measured value (9.9°) reported previously for ECs subjected to 1.6 Pa shear stress for 30 min. The present model is able, for the first time, to cross the boundaries between different length scales in order to provide a global view of potential locations of mechanotransmission.     

Transverse shear modulus and strength of honeycomb cores

The transverse shear mechanical behavior and failure mechanism of aluminum alloy honeycomb cores are investigated by the single block shear test in this paper. The transverse shear deformation process of honeycomb cores may be approximately categorized into four stages, namely elastic deformation, plastic deformation, fracture of cell walls and debonding of honeycomb cores/facesheets. The elastic deformation of unit cell under transverse shear displacement is also investigated by the finite element method, and the result shows that the bending deformation of the cell walls is similar to that of the cantilever beam. In order to precisely predict the equivalent transverse shear modulus and strength, not only shear deformation but also bending deformation of cell walls should be considered. Therefore, in the present paper, the equivalent transverse shear modulus and strength are predicted by application of the cantilever beam theory and thin plate shear buckling theory in conjunction with simplifying assumption as to the displacement in the cores. It is concluded that the contribution of bending deformation of cell walls to equivalent transverse shear modulus and strength is obvious with the decreasing height of cell walls.     

Simulation of Microgravity by Magnetic Levitation and Random Positioning: Effect on Human A431 Cell Morphology

Simulation of weightlessness is a desired replenishment for research in microgravity since access to space flights is limited. In real microgravity conditions, the human epidermoid cell line A431 exhibits specific changes in the actin cytoskeleton resulting ultimately in the rounding-up of cells. This rounding of A431 cells was studied in detail during exposure to Random Positioning Machine (RPM) rotation and magnetic levitation. Random rotation and magnetic levitation induced similar changes in the actin morphology of A431 cells that were also described in real microgravity. A transient process of cell rounding and renewed spreading was observed in time, illustrated by a changing actin cytoskeleton and variation in the presence of focal adhesions. However, side effects of both methods easily can lead to false linking of cellular responses to simulated microgravity. Therefore further characterization of both methods is required.     

The influence of film structure on the interfacial friction in annular two-phase flow under microgravity and normal gravity conditions

Results for the interfacial friction factor and relative interfacial roughness on the gas-liquid interface are reported for an air-water annular flow in a small inner diameter tube (9.53 mm i.d.). The film structure was obtained through processing the time trace signal of film thickness measurements using conductance probes. The interfacial friction factor and the wave height were altered through changing the gravity level and gas Reynolds number. It was found that the wave height decreased with increasing the gas Reynolds number. The wave height in microgravity is less than half of that in normal gravity, while the friction factor was about 10% smaller in microgravity than that in normal gravity. It was shown that the annular two-phase flow friction factor decreased less dramatically as the relative interfacial roughness decreased compared to the single-phase case. It is interesting to note that the interfacial shear stress values at microgravity were very close (or even larger than) those at normal gravity. This was attributed to the thicker substrate at microgravity.     

Martensite and Reverse Transformation during Complete Cycle of Simple Shear of NiTi Shape Memory Alloy

Abstract:  Investigation into the quasi-static simple shear of shape memory alloy (SMA) was carried out. A special grip was designed which allowed replacing compression into the shear process on testing machine. At the same time the shear zone temperature was observed by an infrared camera. The mechanical and thermal characteristics of sheet specimens of NiTi subjected to superelastic shear deformation due to reversible stress-induced phase transformation were clarified. By comparison of the data, the processes of martensite and reverse transformations during the shear deformation were analysed. The investigation shows that stages of the phase transformations during the simple shear process are similar like those observed during tension test. Moreover, during the simple shear deformation no uniform temperature distributions were noticed, especially at higher shear rate, manifesting that the stress-induced phase transformation during the shear process is also inhomogeneous. The thermomechanical properties of the NiTi SMA for various shear rates were investigated. It was found that an increase in the strain rate results in an increase in temperature variation, the shear stresses and the magnitude of hysteresis loops between the forward and reverse transformations.     

聚吡咯的合成及电流变性能的研究

以有机半导体聚吡咯为分散相 ,制备了均相电流变流体 (ER流体 ) ,测试了它在电场下表观粘度、剪切应力等流变学性质的变化。结果表明 ,以 1 溴萘为分散介质的电流变流体在 3kV mm的电场中 ,剪切速率为 12s- 1 时 ,剪切应力达到最大值 10 0 0Pa。剪切应力和表观粘度随温度升高而下降 ,在 5 0℃时 ,最高剪切应力只有 36 0Pa ,而在常温同等条件下 ,剪切应力有 70 0Pa     

See-saw rocking: an in vitro model for mechanotransduction research

In vitro mechanotransduction studies, uncovering the basic science of the response of cells to mechanical forces, are essential for progress in tissue engineering and its clinical application. Many varying investigations have described a multitude of cell responses; however, as the precise nature and magnitude of the stresses applied are infrequently reported and rarely validated, the experiments are often not comparable, limiting research progress. This paper provides physical and biological validation of a widely available fluid stimulation device, a see-saw rocker, as an in vitro model for cyclic fluid shear stress mechanotransduction. This allows linkage between precisely characterized stimuli and cell monolayer response in a convenient six-well plate format. Models of one well were discretized and analysed extensively using computational fluid dynamics to generate convergent, stable and consistent predictions of the cyclic fluid velocity vectors at a rocking frequency of 0.5 Hz, accounting for the free surface. Validation was provided by comparison with flow velocities measured experimentally using particle image velocimetry. Qualitative flow behaviour was matched and quantitative analysis showed agreement at representative locations and time points. Maximum shear stress of 0.22 Pa was estimated near the well edge, and time-average shear stress ranged between 0.029 and 0.068 Pa. Human tenocytes stimulated using the system showed significant increases in collagen and GAG secretion at 2 and 7 day time points. This in vitro model for mechanotransduction provides a versatile, flexible and inexpensive method for the fluid shear stress impact on biological cells to be studied.     

Mechanical behavior and failure process during compressive and shear deformation of honeycomb composite at elevated temperatures

The mechanical behavior and failure mechanism of honeycomb composite consisting of Nomex honeycomb core and 2024Al alloy facesheets were investigated. The compressive and shear deformation behaviors of honeycomb composite were analyzed at temperatures ranged 25–300°C. The compressive and shear strengths of honeycomb composite decreased continuously with increasing temperature up to 300°C. The stress-strain curves obtained from the compressive and shear tests showed that the stress increased to a peak value and then decreased rapidly to a steady state value, which is nearly constant up to failure with increasing strain. The compressive deformation behavior of honeycomb composite was progressed by an elastic and plastic buckling of cell walls, debonding fracture at the interfaces of cell walls, and followed by a fracture of resin layer on cell walls. The shear deformation of honeycomb composite was progressed by an elastic shear deformation, plastic shear deformation, fracture of resin layer on cell walls, and followed by debonding fracture at core/facesheet interfaces. The shear strength of honeycomb composite showed strong anisotropy dependent on the loading direction. The shear strength in longitudinal direction was about 1.4 times higher compared to that in transversal direction due to the different thickness of cell walls mainly loaded during the shear deformation.     

Modeling trabecular bone adaptation to local bending load regulated by mechanosensing osteocytes

Cancellous bone has a complicated three-dimensional porous microstructure that consists of strut-like or plate-like trabeculae. The arrangement of the trabeculae is remodeled throughout the organism’s lifetime to functionally adapt to the surrounding mechanical environment. During bone remodeling, osteocytes buried in the bone matrix are believed to play a pivotal role as mechanosensory cells and help regulate the coupling of osteoclastic bone resorption and osteoblastic bone formation according to the mechanical stimuli. Previously, we constructed a mathematical model of trabecular bone remodeling incorporating cellular mechanosensing and intercellular signal transmission, in which osteocytes are assumed to sense the flow of interstitial fluid as a mechanical stimulus that regulates bone remodeling. Our remodeling simulation could describe the reorientation of a single strut-like trabecula under uniaxial loading. In the present study, to investigate the effects of a bending load on trabecular bone remodeling, we simulated the morphological change in a single trabecula under a cyclic bending load based on our mathematical model. The simulation results showed that the application of the bending load influences not only the formation of the plate-like trabecula but also the changes in trabecular topology. These results suggest the possibility that the characteristic trabecular morphology, such as the strut-like or plate-like form, is determined depending on the local mechanical environment.     

The growth of stem cells within β-TCP scaffolds in a fluid-dynamic environment

A three-dimensional dynamic perfusion system was developed to provide mass transport and nutrient supply to permit the cell proliferation during the long-term culture inside a β-tricalcium phosphate (β-TCP) scaffold. Also the flow field throughout the scaffold was studied. The porous cylindrical scaffold with a central channel was seeded with the sheep mesenchymal stem cells (MSCs). Then the cell-seeded scaffolds were continuously perfused with the complete α-MEM medium by a peristaltic pump for 7, 14 and 28 days, respectively. Histological study showed that the cell proliferation rates were different throughout the whole scaffolds and the different cell coverage was shown in different positions of the scaffold. Unoccupied spaces were found in many macropores. A computational fluid dynamics (CFD) modeling was used to simulate the flow conditions within perfused cell-seeded scaffolds to give an insight into the mechanisms of these cell growth phenomena. Relating the simulation results to perfusion experiments, the even fluid velocity (approximately 0.52 mm/s) and shear stress (approximately 0.0055 Pa) were found to correspond to increased cell proliferation within the cell–scaffold constructs. Flow speeds were between 0.25 and 0.75 mm/s and shear stresses were between 0.003 and 0.008 Pa in approximately 75% of the regions. This method exhibits novel capabilities to compare the results obtained for different perfusion rates or different scaffold microarchitectures. It may allow specific fluid velocities and shear stresses to be determined to optimize the perfusion flow rate, porous scaffold architecture and distribution of in vitro tissue growth.     

Deformation of stem cell nuclei by nanotopographical cues

Cells sense cues in their surrounding microenvironment. These cues are converted into intracellular signals and transduced to the nucleus in order for the cell to respond and adapt its function. Within the nucleus, structural changes occur that ultimately lead to changes in the gene expression. In this study, we explore the structural changes of the nucleus of human mesenchymal stem cells as an effect of topographical cues. We use a controlled nanotopography to drive shape changes to the cell nucleus, and measure the changes with both fluorescence microscopy and a novel light scattering technique. The nucleus changes shape dramatically in response to the nanotopography, and in a manner dependent on the mechanical properties of the substrate. The kinetics of the nuclear deformation follows an unexpected trajectory. As opposed to a gradual shape change in response to the topography, once the cytoskeleton attains an aligned and elongation morphology on the time scale of several hours, the nucleus changes shape rapidly and intensely.     

The effect of simulated microgravity on bacteria from the mir space station

The effects of simulated microgravity on two bacterial isolates, Sphingobacterium thalpophilium and Ralstonia pickettii (formerly Burkholderia pickettii), originally recovered from water systems aboard the Mir space station were examined. These bacteria were inoculated into water, high and low concentrations of nutrient broth and subjected to simulated microgravity conditions. S. thalpophilium (which was motile and had flagella) showed no significant differences between simulated microgravity and the normal gravity control regardless of the method of enumeration and medium. In contrast, for R. pickettii (that was non-motile and lacked flagella), there were significantly higher numbers in high nutrient broth under simulated microgravity compared to normal gravity. Conversely, when R. pikkettii was inoculated into water (i.e., starvation conditions) significantly lower numbers were found under simulated microgravity compared to normal gravity. Responses to microgravity depended on the strain used (e.g., the motile strain exhibited no response to microgravity, while the non-motile strain did), the method of enumeration, and the nutrient concentration of the medium. Under oligotrophic conditions, non-motile cells may remain in geostationary orbit and deplete nutrients in their vicinity, while in high nutrient medium, resources surrounding the cell may be sufficient so that high growth is observed until nutrients becoming limiting.     

酸雨环境下腐蚀RC剪力墙抗震性能试验研究

为研究酸雨环境中RC剪力墙的抗震性能,设计了6榀剪跨比为2.14的RC剪力墙试件,采用人工气候环境模拟技术对其进行酸雨环境加速腐蚀试验,进而进行拟静力试验,分析轴压比和腐蚀程度对RC剪力墙抗震性能的影响。结果表明:在酸雨溶液中的H+和SO₄2+共同侵蚀作用下,试件表面出现析出晶体、侵蚀孔洞、骨料外露等现象,且随酸雨侵蚀循环次数增加腐蚀现象不断加重;随腐蚀程度增加,墙底水平裂缝出现时间提前且裂缝之间间距增加,裂缝宽度变宽,试件承载能力、变形能力和耗能能力均有不同程度的退化,峰值点剪切变形及其占总变形比例减小;随轴压比增加,墙体开裂、屈服和峰值荷载增大,裂缝发展速度减慢,但各试件的变形能力和耗能能力不断降低,峰值点剪切变形及其占总变形比例减小。     

Droplet Deformation and 2-D/3-D Marangoni Flow Phenomena in Droplets Levitated by Electric Fields

An integrated numerical model is presented for free surface phenomena and Marangoni fluid flows in electrically levitated droplets under both terrestrial and microgravity conditions. The model development is based on the boundary element solution of the Maxwell equations simplified for electrostatic levitation applications and the free surface deformation that is primarily caused from the surface Maxwell stresses resulting from the applied electric fields. The electric and free surface model is further integrated with a finite element model for the surface-tension-induced fluid flows in the levitated droplets. Both 2-D and 3-D fluid flow structures may be developed in the electrically levitated droplets depending on the applied laser heating sources. The integrated model is applied to study the electric field distribution, free surface deformation, and 2-D and 3-D internal fluid flow structures in normal and microgravity for single, symmetric two-beam, four-beam, and six-beam laser heating arrangements. Among these arrangements, the six-beam arrangement with equal heating intensity gives the smallest temperature difference and the smallest maximum velocity.     

In vitro and in silico investigations of disc nucleus replacement

Currently, numerous hydrogels are under examination as potential nucleus replacements. The clinical success, however, depends on how well the mechanical function of the host structure is restored. This study aimed to evaluate the extent to and mechanisms by which surgery for nucleus replacements influence the mechanical behaviour of the disc. The effects of an annulus defect with and without nucleus replacement on disc height and nucleus pressure were measured using 24 ovine motion segments. The following cases were considered: intact; annulus incision repaired by suture and glue; annulus incision with removal and re-implantation of nucleus tissue repaired by suture and glue or plug. To identify the likely mechanisms observed in vitro, a finite-element model of a human disc (L4–L5) was employed. Both studies were subjected to physiological cycles of compression and recovery. A repaired annulus defect did not influence the disc behaviour in vitro, whereas additional nucleus removal and replacement substantially decreased disc stiffness and nucleus pressure. Model predictions demonstrated the substantial effects of reductions in replaced nucleus water content, bulk modulus and osmotic potential on disc height loss and pressure, similar to measurements. In these events, the compression load transfer in the disc markedly altered by substantially increasing the load on the annulus when compared with the nucleus. The success of hydrogels for nucleus replacements is not only dependent on the implant material itself but also on the restoration of the environment perturbed during surgery. The substantial effects on the disc response of disruptions owing to nucleus replacements can be simulated by reduced nucleus water content, elastic modulus and osmotic potential.     

Modeled Microgravity Affects Fibroblast Functions Related to Wound Healing

Wound healing is crucial for the survival of an organism. Therefore, in the perspective of space exploration missions, it is important to understand if and how microgravity conditions affect the behavior of the cell populations involved in wound healing and the evolution of the process. Since fibroblasts are the major players in tissue repair, this study was focused on the behavior of fibroblasts in microgravity conditions, modeled by a RCCS. Cell cytoskeleton was studied by immunofluorescence microscopy, the ability to migrate was assessed by microchemotaxis and scratch assay, and the expression of markers of fibroblast activation, angiogenesis, and inflammation was assessed by western blot. Results revealed that after cell exposure to modeled microgravity conditions, a thorough rearrangement of microtubules occurred and α-SMA bundles were replaced by a tight network of faulty and disorganized filaments. Exposure to modeled microgravity induced a decrease in α-SMA and E-CAD expressions. Also, the expression of the pro-angiogenic protein VEGF decreased, while that of the inflammatory signal COX-2 increased. Fibroblast ability to adhere, migrate, and respond to chemoattractants (PRP), closely related to cytoskeleton integrity and membrane junctions, was significantly impaired. Nevertheless, PRP was able to partially restore fibroblast migration.     

Endothelial cell alignment on cyclically-stretched silicone surfaces

Endothelial cells at the interface between the bloodstream and the vessel wall are continuously subjected to mechanical stimulation in vivo, and it widely recognised that such stimulation plays an important role in cardiovascular physiology. Cell deformation is induced by mechanical forces such as cyclic stretch, fluid shear stress, and transmural pressure. Although much of the work in this field has dealt with the effect of fluid shear stress, very little is known about how cyclic forces modulate and alter the morphology of single endothelial cells, and thereafter, how they effect the confluent layer of endothelial cells lining the vessel wall. The aim of this study is to investigate the response of endothelial cells when subjected to substrate deformation of similar magnitude to those experienced in vivo. Human umbilical vein endothelial cells (HUVEC) were cultured on plasma-treated silicone strips and uni-axially cyclically stretched using a custom made mechanical device. Results showed that endothelial cells subject to 10% deformation for as little as 4 h reoriented perpendicular to the stretch direction. In addition, although no integrin coating was applied to the substrate, it was found that plasma-treated silicone provided a cell adhesion substrate comparable to the commonly used collagen type I. Thus the results show that the stretch stimulus alone affects the morphology of endothelial cells. Further studies are required to establish the relative importance of substrate strain vs. fluid flow stimuli.     

考虑剪切变形时薄壁箱梁的挠曲分析

为分析剪切效应对薄壁箱梁受力特性的影响,利用微板的面内剪切及平衡微分方程,分别推导出不考虑和考虑薄壁箱梁各板面内剪切效应的弯曲位移函数。选取剪切效应引起的附加挠度作为广义位移,通过定义的剪切广义力矩及剪切翘曲位移函数,将剪切变形状态从全梁挠曲变形状态中分离出来,作为独立的变形状态进行分析。为满足全截面翘曲应力的自平衡条件,引入两个截面特性参数对广义剪切翘曲位移函数进行了修正。数值算例表明,按该文推导的薄壁箱梁剪切弯曲位移函数计算的两跨连续梁跨中截面应力与实测值及有限元值吻合良好。挠度计算表明:剪切效应使得该箱梁在集中和均布荷载作用下跨中挠度分别增大27%和24%。     

Stress-chain based micromechanics of sand with grain shape effect

The mechanical behaviors of granular media are controlled by grain properties and microstructure. The primary property of granular media is denoted by its grain shape, grain size distribution, stiffness, and interparticle friction. The grain shape itself is of particular importance. Microstructures are formed in the connection paths of contact points between grains. In this paper, the deformation of granular materials with different grain shapes was simulated using two-dimensional DEM under different stress-levels and densities. After analyzing the results, the authors investigated fabric changes. The evolution rule of stress-induced anisotropy and its limitation as well as the existence of a critical state of fabric are revealed.