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Quasi-3D cytoskeletal dynamics of osteocytes under fluid flow.

ABSTRACT: 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.

SUBMITTER: Baik AD

PROVIDER: S-EPMC2965954 | biostudies-literature |

REPOSITORIES: biostudies-literature

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Project description:Complex hemodynamics assessments, as those related to carotid stenosis, are not always easily straightforward due to multifaceted challenges presented by the collateral flow in the Circle of Willis (CoW) and brain flow autoregulation. Advanced computational and benchtop methods to investigate hemodynamics aspects related to such complex flows are often used, however both have limitations and could lead to results which may diverge. In this study we investigated these aspects by performing correlated computational fluid dynamics (CFD) simulations and benchtop experiments in patient specific 3D printed phantoms. To investigate the flow in patients with carotid stenosis, we built two patient specific phantoms which contained the arterial lesion of interest, all main arteries leading to the brain, the CoW and main arteries branching from it. Each phantom was connected to a generic aortic arch. A programmable pump was connected and flow parameters were measured proximal and distal to the lesion and the contralateral arteries. The patient 3D geometry was used to perform a set of CFD simulations where inflow boundary conditions matched the experimental ones. Flow conditions were recorded at the same locations as the experimental setup. Further exploration into the translation from experimental to CFD was also performed by customizing vascular segmentation and physically manipulating arterial compliance properties. We initially observed significant differences between the CFD recordings and the experimental setup. Most of the differences were due to changes in phantom geometry when subjected to physiological pressures and simplistic outflow boundary conditions in the CFD simulations which do not account for pulsatility and nonlinear phenomena. Further work confirms the need for dynamic mesh behavior within CFD simulations attempting to computationally mimic 3D-printed benchtop experiments. Additionally, CFD simulation may benefit from considering geometry specific to a 3D-printed vascular phantom.

Dataset Information

Quasi-3D cytoskeletal dynamics of osteocytes under fluid flow.

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