Bumseok Namgung

Effect of erythrocyte aggregation and flow rate on cell-free layer formation in arterioles

Formation of a cell-free layer is an important dynamic feature of microcirculatory blood flow, which can be influenced by rheological parameters, such as red blood cell aggregation and flow rate. In this study, we investigate the effect of these two rheological parameters on cell-free layer characteristics in the arterioles (20-60 mum inner diameter). For the first time, we provide here the detailed temporal information of the arteriolar cell-free layer in various rheological conditions to better describe the characteristics of the layer variation. The rat cremaster muscle was used to visualize arteriolar flows, and the extent of aggregation was raised by dextran 500 infusion to levels seen in normal human blood. Our results show that cell-free layer formation in the arterioles is enhanced by a combination of flow reduction and red blood cell aggregation. A positive relation (P < 0.005) was found between mean cell-free layer widths and their corresponding SDs for all conditions. An analysis of the frequency and magnitudes of cell-free layer variation from their mean value revealed that the layer deviated with significantly larger magnitudes into the red blood cell core after flow reduction and dextran infusion (P < 0.05). In accordance, the disparity of cell-free layer width distribution found in opposite radial directions from its mean became greater with aggregation in reduced flow conditions. This study shows that the cell-free layer width in arterioles is dependent on both flow rate and red blood cell aggregability, and that the temporal variations in width are asymmetric with a greater excursion into the red blood cell core than toward the vessel wall.

Cell-Free Layer Formation in Small Arterioles at Pathological Levels of Erythrocyte Aggregation

Please cite this paper as: Ong, Jain, Namgung, Woo and Kim (2011). Cell‐Free Layer Formation in Small Arterioles at Pathological Levels of Erythrocyte Aggregation. Microcirculation 18(7), 541–551.

Microfluidic device for sheathless particle focusing and separation using a viscoelastic fluid

Continuous sheathless particle separation with high efficiency is essential for various applications such as biochemical analyses and clinical diagnosis. Here, a novel microfluidic device for highly efficient, sheathless particle separation using an elasticity-dominant non-Newtonian fluid is proposed. Our device consists of two stages: sheathless three-dimensional focusing (1 st stage) and separation (2nd stage). It is designed based on the principle of a viscoelasticity-induced particle lateral migration, which promises precise separation of particles in a microfluidic device. Particles of 5- and 10-μm diameters were all focused at the centerline of a circular channel at the 1st stage and successfully separated at the 2nd stage with an efficiency of ∼99.9% using size-based lateral migration of particles induced by the viscoelasticity of the medium. We also demonstrated the capability of our device for separation of blood cells into multiple fractions. The tunability of separable particle size could be achieved by changing the viscoelastic property of the medium and flow rate.

A review of numerical methods for red blood cell flow simulation

In this review, we provide an overview of the simulation techniques employed for modelling the flow of red blood cells (RBCs) in blood plasma. The scope of this review omits the fluid modelling aspect while focusing on other key components in the RBC–plasma model such as (1) describing the RBC deformation with shell-based and spring-based RBC models, (2) constitutive models for RBC aggregation based on bridging theory and depletion theory and (3) additional strategies required for completing the RBC–plasma flow model. These include topics such as modelling fluid–structure interaction with the immersed boundary method and boundary integral method, and updating the variations in multiphase fluid property through the employment of index field methods. Lastly, we summarily discuss the current state and aims of RBC modelling and suggest some research directions for the further development of this field of modelling.

Hybrid capillary-inserted microfluidic device for sheathless particle focusing and separation in viscoelastic flow

A novel microfluidic device which consists of two stages for particle focusing and separation using a viscoelastic fluid has been developed. A circular capillary tube was used for three-dimensional particle pre-alignment before the separation process, which was inserted in a polydimethylsiloxane microchannel. Particles with diameters of 5 and 10 μm were focused at the centerline in the capillary tube, and the location of particles was initialized at the first bifurcation. Then, 5 and 10 μm particles were successfully separated in the expansion region based on size-dependent lateral migration, with ∼99% separation efficiency. The proposed device was further applied to separation of MCF-7 cells from leukocytes. Based on the cell size distribution, an approximate size cutoff for separation was determined to be 16 μm. At 200 μl/min, 94% of MCF-7 cells were separated with the purity of ∼97%. According to the trypan blue exclusion assay, high viability (∼90%) could be achieved for the separated MCF-7 cells. The use of a commercially available capillary tube enables the device to be highly versatile in dealing with particles in a wide size range by using capillary tubes with different inner diameters.

A comparative study of histogram-based thresholding methods for the determination of cell-free layer width in small blood vessels

We have recently proposed a computer-based method utilizing a thresholding algorithm (the Otsu method) to provide a convenient way of measuring the cell-free layer width in vivo and in vitro. However, this method does not seem to be a universal method that can be applied to all microvascular studies. Thus, we examined four different histogram-based thresholding algorithms (Otsu, intermode, minimum and second peak) to provide a technical suggestion on the selection of a suitable thresholding algorithm for the cell-free layer measurement. All the measurements were taken in microvascular flows in the rat cremaster muscle recorded with a high-speed camera. The width of the cell-free layer manually measured was compared with that determined by the automated method utilizing the four thresholding algorithms. With our experimental system, results showed that the cell-free layer width determined by the minimum algorithm was in best accordance with the manual measurement. We concluded that the accuracy of the automated methods for determination of the cell-free layer width would depend on the image quality, in particular on the contrast between the red blood cell core and background, which might differ due to the different microscopic setup. Therefore, one may need to examine several appropriate thresholding methods when selecting the best suitable algorithm for the experimental conditions.

An automated method for cell-free layer width determination in small arterioles

Histogram-based thresholding techniques utilized for cell-free layer width measurement in arteriolar flow may produce an overestimation of the layer width since they do not consider faint shaded regions near the vessel wall as part of the erythrocyte column. To address this problem, we developed a new method for detecting the boundary of the erythrocyte column based on an edge detection algorithm. This automated method (grayscale method) provides local detections of the inner vessel wall as well as the boundary between the cell-free layer and the erythrocyte column without binarization of grayscale images. The cell-free layer width measurements using the grayscale method and existing techniques (minimum method and Otsus method) were compared with those determined manually in arteriolar flows of the rat cremaster muscle. In the absence of the shaded regions, values obtained by the grayscale method and minimum method were statistically in good agreement with the manual method but not in the case of Otsus method. When the faint shaded regions were present, the grayscale method appeared to produce more accurate results than the minimum method and Otsus method.

Effect of Input Waveform Pattern and Large Blood Vessel Existence on Destruction of Liver Tumor Using Radiofrequency Ablation: Finite Element Analysis

Much research has been directed at improving the effectiveness of the radiofrequency (RF) ablation of hepatocellular carcinomas. In that point of view, this study was performed to provide comprehensive information of the relation between RF waveforms and thermodynamic response of the tissue with the consideration of four different types of RF waveforms (half-sine, half-square, half-exponential, and damped-sine) to maximize the amount of tumor tissue removed while maintaining the advantages of RF ablation. For the aim of this study, finite element models incorporating results from previous numerical models were used and validated with ex vivo experiments. From analyses of the entire results, we concluded that this study may prove valuable as a first step in providing comprehensive information of the relation between various RF waveforms and thermodynamic responses within the tissue during the RF ablation process. This study may also contribute toward studies to determine an optimum RF waveform capable of maximizing the amount of tumor tissue removed while maintaining the advantages of RF ablation.

Effect of erythrocyte aggregation at pathological levels on NO/O2 transport in small arterioles

This study examined the effects of red blood cell (RBC) aggregation at pathological levels on NO/O2 transport in small arterioles. Transient gas diffusion simulations were performed with in vivo cell-free layer (CFL) widths data obtained from arteriolar flows in the rat cremaster muscle. The CFL data were measured at physiological and pathological levels of aggregation under reduced flow conditions (pseudoshear rate = 31.4 ± 10.5 s-1). Our results showed that the mean peak NO concentration significantly decreased with increasing the aggregation level from non-aggregating to normal-aggregating (P < 0.05) and to hyper-aggregating (P < 0.01) conditions. In contrast, the partial O2 pressure (PO2) in pathological aggregating conditions significantly increased from those under non-aggregating (P < 0.001) and normal-aggregating (P < 0.05) conditions. Although the NO scavenging by RBCs could be impaired with a thicker CFL at higher levels of aggregation, the overall decrease in NO production due to reduction of wall shear stress with the thicker CFL dominantly limited the NO availability in tissue. On the other hand, the O2 availability in tissue increased due to the relatively high core hematocrit in the blood lumen with the thicker CFL.

Two-dimensional transient model for prediction of arteriolar NO/O2 modulation by spatiotemporal variations in cell-free layer width.

Despite the significant roles of the cell-free layer (CFL) in balancing nitric oxide (NO) and oxygen (O2) bioavailability in arteriolar tissue, many previous numerical approaches have relied on a one-dimensional (1-D) steady-state model for simplicity. However, these models are unable to demonstrate the influence of spatiotemporal variations in the CFL on the NO/O2 transport under dynamic flow conditions. Therefore, the present study proposes a new two-dimensional (2-D) transient model capable of predicting NO/O2 transport modulated by the spatiotemporal variations in the CFL width. Our model predicted that NO bioavailability was inversely related to the CFL width as expected. The enhancement of NO production by greater wall shear stress with a thinner CFL could dominate the diffusion barrier role of the CFL. In addition, NO/O2 availability along the vascular wall was inhomogeneous and highly regulated by dynamic changes of local CFL width variation. The spatial variations of CFL widths on opposite sides of the arteriole exhibited a significant inverse relation. This asymmetric formation of CFL resulted in a significantly imbalanced NO/O2 bioavailability on opposite sides of the arteriole. The novel integrative methodology presented here substantially highlighted the significance of spatiotemporal variations of the CFL in regulating the bioavailability of NO/O2, and provided further insight about the opposing effects of the CFL on arteriolar NO production.