Ken-ichi Tsubota

Confocal micro-PIV measurements of three-dimensional profiles of cell suspension flow in a square microchannel

A detailed measurement of the blood flow velocity profile in microchannels in vitro is fundamental to better understand the biomechanics of microcirculation. Therefore it is very important to determine the influence of suspended blood cells on the flow behaviour with high accuracy and spatial resolution. We measured the flow of blood cells suspended in a physiological fluid within a square microchannel using a confocal particle image velocimetry (PIV) system and compared it to pure water. This emerging technology combines a conventional PIV system with a spinning confocal microscope and has the ability to obtain high-resolution images and three-dimensional (3D) optical section velocity measurements. The good agreement obtained between the measured and estimated results suggests that macroscale flow theory can be used to predict the flow behaviour of a homogeneous fluid within a 100 µm square microchannel. Our results also demonstrated the potential of the confocal system for generating 3D profiles and consequently obtaining detailed information on microscale effects in microchannels using both homogeneous and non-homogeneous fluids, such as a suspension of blood cells. Furthermore, the results obtained from our confocal micro-PIV system show the ability of this system to measure velocities up to 0.52 mm s−1 in a blood cell suspension fluid.

Trabecular Surface Remodeling Simulation for Cancellous Bone Using Microstructural Voxel Finite Element Models

A computational simulation method for three-dimensional trabecular surface remodeling was proposed, using voxel finite element models of cancellous bone, and was applied to the experimental data. In the simulation, the trabecular microstructure was modeled based on digital images, and its morphological changes due to surface movement at the trabecular level were directly expressed by removing/adding the voxel elements from/to the trabecular surface. A remodeling simulation at the single trabecular level under uniaxial compressive loading demonstrated smooth morphological changes even though the trabeculae were modeled with discrete voxel elements. Moreover, the trabecular axis rotated toward the loading direction with increasing stiffness, simulating functional adaptation to the applied load. In the remodeling simulation at the trabecular structural level, a cancellous bone cube was modeled using a digital image obtained by microcomputed tomography (microCT), and was uniaxially compressed. As a result, the apparent stiffness against the applied load increased by remodeling, in which the trabeculae reoriented to the loading direction. In addition, changes in the structural indices of the trabecular architecture coincided qualitatively with previously published experimental observations. Through these studies, it was demonstrated that the newly proposed voxel simulation technique enables us to simulate the trabecular surface remodeling and to compare the results obtained using this technique with the in vivo experimental data in the investigation of the adaptive bone remodeling phenomenon.

Functional adaptation of cancellous bone in human proximal femur predicted by trabecular surface remodeling simulation toward uniform stress state.

Two-dimensional simulation of trabecular surface remodeling was conducted for a human proximal femur to investigate the structural change of cancellous bone toward a uniform stress state. Considering that a local mechanical stimulus plays an important role in cellular activities in bone remodeling, local stress nonuniformity was assumed to drive trabecular structural change to seek a uniform stress state. A large-scale pixel-based finite element model was used to simulate structural changes of individual trabeculae over the entire bone. As a result, the initial structure of trabeculae changed from isotropic to anisotropic due to trabecular microstructural changes caused by surface remodeling according to the mechanical environment in the proximal femur. Under a single-loading condition, it was shown that the apparent structural property evaluated by fabric ellipses corresponded to the apparent stress state in cancellous bone. As is observed in the actual bone, a distributed trabecular structure was obtained under a multiple-loading condition. Through these studies, it was concluded that trabecular surface remodeling toward a local uniform stress state at the trabecular level could naturally bring about functional adaptation phenomenon at the apparent tissue level. The proposed simulation model would be capable of providing insight into the hierarchical mechanism of trabecular surface remodeling at the microstructural level up to the apparent tissue level.

Computer simulation of trabecular remodeling in human proximal femur using large-scale voxel FE models : Approach to understanding Wolff's law

Ever since Julius Wolff proposed the law of bone transformation in the 19th century, it has been widely known that the trabecular structure of cancellous bone adapts functionally to the loading environment. To understand the mechanism of Wolffs law, a three-dimensional (3D) computer simulation of trabecular structural changes due to surface remodeling was performed for a human proximal femur. A large-scale voxel finite element model was constructed to simulate the structural changes of individual trabeculae over the entire cancellous region. As a simple remodeling model that considers bone cellular activities regulated by the local mechanical environment, nonuniformity of local stress was assumed to drive the trabecular surface remodeling to seek a uniform stress state. Simulation results demonstrated that cell-scale ( approximately 10microm) remodeling in response to mechanical stimulation created complex 3D trabecular structures of the entire bone-scale ( approximately 10cm), as illustrated in the reference of Wolff. The bone remodeling reproduced the characteristic anisotropic structure in the coronal cross section and the isotropic structures in other cross sections. The principal values and axes of a structure characterized by fabric ellipsoids corresponded to those of the apparent stress of the structure. The proposed large-scale computer simulation indicates that in a complex mechanical environment of a hierarchical bone structure of over 10(4) length scale (from approximately 10microm to approximately 10cm), a simple remodeling at the cellular/trabecular levels creates a highly complex and functional trabecular structure, as characterized by bone density and orientation.

Particle method for computer simulation of red blood cell motion in blood flow.

A particle method for the computer simulation of blood flow was proposed to analyze the motion of a deformable red blood cell (RBC) in flowing blood plasma. The RBC and plasma were discretized by particles that have the characteristics of an elastic membrane and a viscous fluid, respectively. The membrane particles were connected to their neighboring membrane particles by springs, and the motion of the particles was determined on the basis of the minimum energy principle. The incompressible flow of plasma that was expressed by the motion of the fluid particles was determined by the moving-particle semi-implicit (MPS) method. The RBC motion and plasma flow were weakly coupled. The two-dimensional simulation of blood flow between parallel plates demonstrated the capability of the proposed method to express the blood flow phenomena observed in experiments, such as the downstream motion of the RBC and the deformation of the RBC into a parachute shape.

Computational study on effect of red blood cells on primary thrombus formation.

The primary thrombus formation is a critical phenomenon both physiologically and pathologically. It has been seen that various mechanical factors are involved the regulation of primary thrombus formation through a series of physiological and biochemical processes, including blood flow and intercellular molecular bridges. However, it has not been fully understood how the existence of red blood cells contributes to the process of platelet thrombus formation. We computationally analyzed the motions of platelets in plasma layer above which red blood cells flow assuming a background simple shear flow of a shear rate of 1000 s(-1) using Stokesian dynamics. In the computation, fluid mechanical interactions between platelets and red blood cells were taken into account together with the binding forces via plasma proteins between two platelets and between a platelet and injured vessel wall. The process of the platelets aggregation was significantly dependent on whether red blood cells were present. When red blood cells were absent, the aggregate formed grew more vertically compared to the case with red blood cells. Conversely, when red blood cells were present, the aggregate spread more horizontally because the red blood cells constrained the vertical growth when the height of the aggregate reached the level of the red blood cells. Our results suggest that red blood cells mechanically play a significant role in primary thrombus formation, which accelerates the horizontal spread of the thrombus, and point out the necessity of considering the presence of red blood cells when investigating the mechanism of thrombus formation.

Hemodynamic Analysis of Microcirculation in Malaria Infection

Malaria-infected red blood cells (IRBCs) show various changes in mechanical properties. IRBCs lose their deformability and develop properties of cytoadherence and rosetting. To clarify how these changes advance microvascular occlusion, we need qualitative and quantitative information on hemodynamics in malaria infection, including the interaction among IRBCs, healthy RBCs, and endothelial cells. We developed a numerical model of blood flow with IRBCs based on conservation laws of fluid dynamics. The deformability and adhesive property of IRBCs were simply modeled using springs governed by Hook’s law. Our model could express the basic behavior of IRBCs, including the rolling motion due to cytoadhesive interaction with endothelial cells and complex interaction with healthy RBCs. We confirmed that these types of interactions significantly increase the flow resistance, particularly when knobs develop.

Effects of a fixation screw on trabecular structural changes in a vertebral body predicted by remodeling simulation

AbstractComputational simulation of trabecular surface remodeling was conducted to investigate the effects of a spinal fixation screw on trabecular structural changes in a vertebral body. By using voxel-based finite elements, computational models of the bone and screw were constructed in two structural scales of a vertebral body with an implanted screw and a bone–screw interface. In the vertebral body, the implantation of the fixation screw caused changes in the mechanical environment in cancellous bone, leading to trabecular structural changes at the cancellous level. The effects of the screw on trabecular orientation were greater in the regions above and below the screw than in those in front of the screw. In the case of the bone–screw interface, trabecular structural changes depended on the direction of load applied to the screw. It was suggested that the bone resorption predicted in the pull-out loading case is a candidate cause of the loosening of the screw. The results indicate that the effects of the implanted screw on trabecular structural changes are more important for longer-term fixation.

Simulation of platelet adhesion and aggregation regulated by fibrinogen and von Willebrand factor

We propose a method to analyze platelet adhesion and aggregation computationally, taking into account the distinct properties of two plasma proteins, von Willebrand factor (vWF) and fibrinogen (Fbg). In this method, the hydrodynamic interactions between platelet particles under simple shear flow were simulated using Stokesian dynamics based on the additivity of velocities. The binding force between particles mediated by vWF and Fbg was modeled using the Voigt model. Two Voigt models with different properties were introduced to consider the distinct behaviors of vWF and Fbg. Our results qualitatively agreed with the general observation of a previous in-vitro experiment, thus demonstrating that the significant development of thrombus formation in height requires not only vWF, but also Fbg. This agreement of simulation and experimental results qualitatively validates our model and suggests that consideration of the distinct roles of vWF and Fbg is essential to investigate the physiological and pathophysiological mechanisms of thrombus formation using a computational approach.

Gait Analysis for Detecting a Leg Accident with an Accelerometer

We analyzed an acceleration pattern during natural walking and walking hampered by wearing weights to predict falls. These two types of walking could be distinguished with a peak at the half of the principal frequency of the gait in anterior movement. We measured walking while wearing each individual hampering weight and then analyzed. The peak was bigger when a subject wore the hampering weight that was restrictive and made the body unbalanced. As leg accidents are considered to occur with a change of the body balance, our system may be used to detect a leg accident by checking the peak at the half of the principal frequency of the gait in anterior movement