Energy-Mediated Machinery Drives Cellular Mechanical Allostasis.

Allostasis is a fundamental biological process through which living organisms achieve stability via physiological or behavioral changes to protect against internal and external stresses, and ultimately better adapt to the local environment. However, an full understanding of cellular-level allostasis is far from developed. By employing an integrated micromechanical tool capable of applying controlled mechanical stress on an individual cell and simultaneously reporting dynamic information of subcellular mechanics, individual cell allostasis is observed to occur through a biphasic process; cellular mechanics tends to restore to a stable state through a mechanoadaptative process with excitative biophysical activity followed by a decaying adaptive phase. Based on these observations, it is found that cellular allostasis occurs through a complex balance of subcellular energy and cellular mechanics; upon a transient and local physical stimulation, cells trigger an allostatic state that maximizes energy and overcomes a mechanical "energy barrier" followed by a relaxation state that reaches its mechanobiological stabilization and energy minimization. Discoveries of energy-driven cellular machinery and conserved mechanotransductive pathways underscore the critical role of force-sensitive cytoskeleton equilibrium in cellular allostasis. This highlight the biophysical origin of cellular mechanical allostasis, providing subcellular methods to understand the etiology and progression of certain diseases or aging.

Effects of mechanical loading on cortical defect repair using a novel mechanobiological model of bone healing.

Mechanical loading is an important aspect of post-surgical fracture care. The timing of load application relative to the injury event may differentially regulate repair depending on the stage of healing. Here, we used a novel mechanobiological model of cortical defect repair that offers several advantages including its technical simplicity and spatially confined repair program, making effects of both physical and biological interventions more easily assessed. Using this model, we showed that daily loading (5N peak load, 2Hz, 60 cycles, 4 consecutive days) during hematoma consolidation and inflammation disrupted the injury site and activated cartilage formation on the periosteal surface adjacent to the defect. We also showed that daily loading during the matrix deposition phase enhanced both bone and cartilage formation at the defect site, while loading during the remodeling phase resulted in an enlarged woven bone regenerate. All loading regimens resulted in abundant cellular proliferation throughout the regenerate and fibrous tissue formation directly above the defect demonstrating that all phases of cortical defect healing are sensitive to physical stimulation. Stress was concentrated at the edges of the defect during exogenous loading, and finite element (FE)-modeled longitudinal strain (εzz) values along the anterior and posterior borders of the defect (~2200µÎµ) was an order of magnitude larger than strain values on the proximal and distal borders (~50-100µÎµ). It is concluded that loading during the early stages of repair may impede stabilization of the injury site important for early bone matrix deposition, whereas loading while matrix deposition and remodeling are ongoing may enhance stabilization through the formation of additional cartilage and bone.

Quantifying the ultrastructure of carotid arteries using high-resolution micro-diffusion tensor imaging-comparison of intact versus open cut tissue.

Diffusion magnetic resonance imaging (dMRI) can provide insights into the microstructure of intact arterial tissue. The current study employed high magnetic field MRI to obtain ultra-high resolution dMRI at an isotropic voxel resolution of 117 µm3 in less than 2 h of scan time. A parameter selective single shell (128 directions) diffusion-encoding scheme based on Stejskel-Tanner sequence with echo-planar imaging (EPI) readout was used. EPI segmentation was used to reduce the echo time (TE) and to minimise the susceptibility-induced artefacts. The study utilised the dMRI analysis with diffusion tensor imaging (DTI) framework to investigate structural heterogeneity in intact arterial tissue and to quantify variations in tissue composition when the tissue is cut open and flattened. For intact arterial samples, the region of interest base comparison showed significant differences in fractional anisotropy and mean diffusivity across the media layer (p < 0.05). For open cut flat samples, DTI based directionally invariant indices did not show significant differences across the media layer. For intact samples, fibre tractography based indices such as calculated helical angle and fibre dispersion showed near circumferential alignment and a high degree of fibre dispersion, respectively. This study demonstrates the feasibility of fast dMRI acquisition with ultra-high spatial and angular resolution at 7 T. Using the optimised sequence parameters, this study shows that DTI based markers are sensitive to local structural changes in intact arterial tissue samples and these markers may have clinical relevance in the diagnosis of atherosclerosis and aneurysm.

Immersed boundary-finite element model of fluid-structure interaction in the aortic root.

It has long been recognized that aortic root elasticity helps to ensure efficient aortic valve closure, but our understanding of the functional importance of the elasticity and geometry of the aortic root continues to e-volve as increasingly detailed in vivo imaging data become available. Herein, we describe a fluid-structure interaction model of the aortic root, including the aortic valve leaflets, the sinsuses of Valsalva, the aortic annulus, and the sinotubular junction, that employs a version of Peskin's immersed boundary (IB) method with a finite element (FE) description of the structural elasticity. As in earlier work, we use a fiber-based model of the valve leaflets, but this study extends earlier IB models of the aortic root by employing an incompressible hyperelastic model of the mechanics of the sinuses and ascending aorta using a constitutive law fit to experimental data from human aortic root tissue. In vivo pressure loading is accounted for by a backward displacement method that determines the unloaded configurations of the root model. Our model yields realistic cardiac output at physiological pressures, with low transvalvular pressure differences during forward flow, minimal regurgitation during valve closure, and realistic pressure loads when the valve is closed during diastole. Further, results from high-resolution computations indicate that although the detailed leaflet and root kinematics show some grid sensitivity, our IB model of the aortic root nonetheless produces essentially grid-converged flow rates and pressures at practical grid spacings for the high-Reynolds number flows of the aortic root. These results thereby clarify minimum grid resolutions required by such models when used as stand-alone models of the aortic valve as well as when used to provide models of the outflow valves in models of left ventricular fluid dynamics.

A numerical framework for the mechanical analysis of dual-layer stents in intracranial aneurysm treatment.

Dual-layer stents and multi-layer stents represent a new paradigm in endovascular interventions. Multi-layer stents match different stent designs in order to offer auxiliary functions. For example, dual-layer stents used in the endovascular treatment of intracranial aneurysms, like the FRED(TM) (MicroVention, CA) stent, combine a densely braided inner metallic mesh with a loosely braided outer mesh. The inner layer is designed to divert blood flow, whereas the outer one ensures microvessels branching out of the main artery remain patent. In this work, the implemented finite element (FE) analysis identifies the key aspects of dual-stent mechanics. In particular, dual-layer stents used in the treatment of intracranial aneurysms require the ability to conform to very narrow passages in their closed configuration, while at the same time they have to provide support and stability once deployed. This study developed a numerical framework for the analysis of dual-layer stents for endovascular intracranial aneurysm treatment. Our results were validated against analytical methods. For the designs considered, we observed that foreshortening was in average 37.5%±2.5%, and that doubling the number of wires in the outer stent increased bending moment by 23%, while halving the number of wires of the inner stent reduced von Mises stress by 2.3%. This framework can be extended to the design optimization of multi-layer stents used in other endovascular treatments.

Osteoblast-derived paracrine factors regulate angiogenesis in response to mechanical stimulation.

Angiogenesis is a process by which new blood vessels emerge from existing vessels through endothelial cell sprouting, migration, proliferation, and tubule formation. Angiogenesis during skeletal growth, homeostasis and repair is a complex and incompletely understood process. As the skeleton adapts to mechanical loading, we hypothesized that mechanical stimulation regulates "osteo-angio" crosstalk in the context of angiogenesis. We showed that conditioned media (CM) from osteoblasts exposed to fluid shear stress enhanced endothelial cell proliferation and migration, but not tubule formation, relative to CM from static cultures. Endothelial cell sprouting was studied using a dual-channel collagen gel-based microfluidic device that mimics vessel geometry. Static CM enhanced endothelial cell sprouting frequency, whereas loaded CM significantly enhanced both frequency and length. Both sprouting frequency and length were significantly enhanced in response to factors released from osteoblasts exposed to fluid shear stress in an adjacent channel. Osteoblasts released angiogenic factors, of which osteopontin, PDGF-AA, IGBP-2, MCP-1, and Pentraxin-3 were upregulated in response to mechanical loading. These data suggest that in vivo mechanical forces regulate angiogenesis in bone by modulating "osteo-angio" crosstalk through release of paracrine factors, which we term "osteokines".

Imaging and finite element analysis: a methodology for non-invasive characterization of aortic tissue.

Characterization of the mechanical properties of arterial tissues usually involves an invasive procedure requiring tissue removal. In this work we propose a non-invasive method to perform a biomechanical analysis of cardiovascular aortic tissue. This method is based on combining medical imaging and finite element analysis (FEA). Magnetic resonance imaging (MRI) was chosen since it presents relatively low risks for human health. A finite element model was created from the MRI images and loaded with systolic physiological pressures. By means of an optimization routine, the structural material properties were changed until average strains matched those measured by MRI. The method outlined in this work produced an estimate of the in situ properties of cardiovascular tissue based on non-invasive image datasets and finite element analysis.

Fibre orientation of fresh and frozen porcine aorta determined non-invasively using diffusion tensor imaging.

Diffusion tensor imaging analysis was applied to fresh and frozen porcine aortas in order to determine fibre orientation. Fresh and stored frozen porcine aortas were imaged in a 7 T scanner with a diffusion weighted spin echo sequence (six gradient directions, matrix 128×128 pixels, 2.8 cm×2.8 cm field of view). The images were taken for different b values, ranging from 200 s/mm(2) to 1600 s/mm(2). For each dataset the diffusion tensor was evaluated, fractional anisotropy (FA) maps were calculated, and the fibres mapped. The arterial fibres resulting were postprocessed and their fibre angle evaluated. The FA maps, the dominant fibre angle, and the fibre pattern in the arterial wall thickness were compared in the fresh and in the stored frozen aortas. The technique was able to determine a fibre pattern in the fresh healthy aorta that is in accordance with the data available in literature and to identify an alteration in the fibre pattern caused by freezing. This study shows that this technique has potential for studying fibre orientation and fibre distribution in humans and could be further developed to diagnose fibre alterations due to cardiovascular diseases. In fact, our results suggest that DTI has the potential to determine the fibrous structure of arteries non-invasively. This capability could be further developed to study the natural remodelling of the aorta in vivo due to age and/or gender or to obtain information on aortic diseases at an early stage of their evolution.