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| Analysis of TNF-α-induced Leukocyte Adhesion to Vascular Endothelial Cells Regulated by Fluid Shear Stress Using Microfluidic Chip-based Technology |
| LI Yuan, YANG De-yu, LIAO Juan, GONG Fang, HE Ping, LIU Bei-zhong |
| Central Laboratory of Yong Chuan Hospital, Chongqing Medical University, Chongqing 402160, China |
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Abstract This paper aims to the research of the impact of fluid shear stress on the adhesion between vascular endothelial cells and leukocyte induced by tumor necrosis factor-α(TNF-α) by microfliudic chip technology. Microfluidic chip was fabricated by soft lithograph; Endothelial microfluidic chip was constructed by optimizing types of the extracellular matrix proteins modified in the microchannel and cell incubation time; human umbilical vein endothelial cells EA.Hy926 lined in the microchannel were exposed to fluid shear stress of 1.68 dynes/cm2 and 8.4 dynes/cm2 respectively. Meanwhile, adhesion between EA.Hy926 cells and leukocyte was induced by TNF-αunder a flow condition. EA. Hy926 cell cultured in the static condition was used as control group. The numbers of fluorescently-labeled leukocyte in microchannel were counted to quantize the adhesion level between EA. Hy926 cells and leukocyte; cell immunofluorescence technique was used to detect the intercellular adhesion molecule (ICAM-1) expression. The constructed endothelial microfluidic chip can afford to the fluid shear stress and respond to exogenous stimulus of TNF-α; compared with the adhesion numbers of leukocyte in control group, adhesion between EA. Hy926 cells exposed to low fluid shear stress and leukocyte was reduced under the stimulus of TNF-α at a concentration of 10 ng/ml(P<0.05); leukocyte adhesion with EA. Hy926 cells exposed to high fluid shear stress was reduced significantly than EA. Hy926 cells in control group and EA.1Hy926 cells exposed to low fluid shear stress(P<0.01); the regulation mechanism of fluid shear stress to the adhesion between EA. Hy926 cells and leukocyte induced by TNF-αwas through the way of ICAM-1. The endothelial microfluidic chip fabricated in this paper could be used to study the functions of endothelial cell in vitro and provide a new technical platform for exploring the pathophysiology of the related cardiovascular system diseases under a flow environment.
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Received: 15 August 2015
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Corresponding Authors:
LI Yuan. E-mail: liyuan_1985@163.com
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