Quasi-3D Cytoskeletal Dynamics of Osteocytes under Fluid Flow

Osteocytes respond to dynamic fluid shear loading by activating various biochemical pathways, mediating a dynamic process of bone formation and resorption. Whole-cell deformation and regional deformation of the cytoskeleton may be able to directly regulate this process. Attempts to image cellular deformation by conventional microscopy techniques have been hindered by low temporal or spatial resolution. In this study, we developed a quasi-three-dimensional microscopy technique that enabled us to simultaneously visualize an osteocyte's traditional bottom-view profile and a side-view profile at high temporal resolution. Quantitative analysis of the plasma membrane and either the intracellular actin or microtubule (MT) cytoskeletal networks provided characterization of their deformations over time. Although no volumetric dilatation of the whole cell was observed under flow, both the actin and MT networks experienced primarily tensile strains in all measured strain components. Regional heterogeneity in the strain field of normal strains was observed in the actin networks, especially in the leading edge to flow, but not in the MT networks. In contrast, side-view shear strains exhibited similar subcellular distribution patterns in both networks. Disruption of MT networks caused actin normal strains to decrease, whereas actin disruption had little effect on the MT network strains, highlighting the networks' mechanical interactions in osteocytes.

Linear spatio-temporal instability analysis of ice growth under a falling water film

A linear spatio-temporal stability analysis is conducted for the ice growth under a falling water film along an inclined ice plane. The full system of linear stability equations is solved by using the Chebyshev collocation method. By plotting the boundary curve between the linear absolute and convective instabilities (AI/CI) of the ice mode in the parameter plane of the Reynolds number and incline angle, it is found that the linear absolute instability exists and occurs above a minimum Reynolds number and below a maximum inclined angle. Furthermore, by plotting the critical Reynolds number curves with respect to the inclined angle for the downstream and upstream branches, the convectively unstable region is determined and divided into three parts, one of which has both downstream and upstream convectively unstable wavepackets and the other two have only downstream or upstream convectively unstable wavepacket. Finally, the effect of the Stefan number and the thickness of the ice layer on the AI/CI boundary curve is investigated.

An ATP-dependent mechanism mediates intercellular calcium signaling in bone cell network under single cell nanoindentation

To investigate the roles of intercellular gap junctions and extracellular ATP diffusion in bone cell calcium signaling propagation in bone tissue, in vitro bone cell networks were constructed by using microcontact printing and self-assembled monolayer technologies. In the network, neighboring cells were interconnected through functional gap junctions. A single cell at the center of the network was mechanically stimulated by using an AFM nanoindenter. Intracellular calcium ([Ca2+](i)) responses of the bone cell network were recorded and analyzed. In the untreated groups, calcium propagation from the stimulated cell to neighboring cells was observed in 40% of the tests. No significant difference was observed in this percentage when the intercellular gap junctions were blocked. This number, however, decreased to 10% in the extracellular ATP-pathway-blocked group. When both the gap junction and ATP pathways were blocked, intercellular calcium waves were abolished. When the intracellular calcium store in ER was depleted, the indented cell can generate calcium transients, but no [Ca2+](i) signal can be propagated to the neighboring cells. No [Ca2+](i) response was detected in the cell network when the extracellular calcium source was removed. These findings identified the biochemical pathways involved in the calcium signaling propagation in bone cell networks. Published by Elsevier Ltd.

Intercellular calcium wave propagation in linear and circuit-like bone cell networks

In the present study, the mechanism of intercellular calcium wave propagation in bone cell networks was identified. By using micro-contact printing and self-assembled monolayer technologies, two types of in vitro bone cell networks were constructed: open-ended linear chains and looped hexagonal networks with precisely controlled intercellular distances. Intracellular calcium responses of the cells were recorded and analysed when a single cell in the network was mechanically stimulated by nano-indentation. The looped cell network was shown to be more efficient than the linear pattern in transferring calcium signals from cell to cell. This phenomenon was further examined by pathway-inhibition studies. Intercellular calcium wave propagation was significantly impeded when extracellular adenosine triphosphate (ATP) in the medium was hydrolysed. Chemical uncoupling of gap junctions, however, did not significantly decrease the transferred distance of the calcium wave in the cell networks. Thus, it is extracellular ATP diffusion, rather than molecular transport through gap junctions, that dominantly mediates the transmission of mechanically elicited intercellular calcium waves in bone cells. The inhibition studies also demonstrated that the mechanical stimulation-induced calcium responses required extracellular calcium influx, whereas the ATP-elicited calcium wave relied on calcium release from the calcium store of the endoplasmic reticulum.

Numerical Study on the Spatial and Temporal Characteristics of Water Quality in the Taihu Lake of China

Taihu Lake is the third largest fresh water lake in China. With the fast economic development, abundant industrial and agricultural waste water has been discharged into Taihu Lake, causing the eutrophication of the water quality, which greatly affected the water utility. In the past decades, the treatment of Taihu Lake has witnessed limited success. Therefore, it is practically and theoretically significant to study the eutrophication of Taihu Lake. This research has focused on the issue of water quality including the characteristics of spatial and temporal distributions, and the rules of nutrient diffusion in the Taihu lake area. Based on the monitoring data, the basis distribution characteristics of water quality in Taihu Lake are analyzed. Comparing Taihu Lake with other Lakes shows that one important reason for Taihu eutrophication is the long period of water retention. A transporting and diffusing model of Taihu nutrient is developed by combining with the hydrodynamics model. Using the model, the concentration field of the total phosphorus (TP) and the influence of wind-driven current are numerically investigated, which leads to the conclusion that the flow field has a great influence on the spatial and temporal distributions of TP in Taihu Lake. Furthermore, the effect for improving the water quality by the project of water diversion from the Yangtze River to Taihu Lake was analyzed by simulation. The results demonstrate that short-term water diversion cannot improve the water quality of the heavily-polluted Meiliang Bay and the western bank areas of Taihu Lake.