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.Microflu...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/cm^2 and 8.4 dynes/cm^2 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.展开更多
文摘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/cm^2 and 8.4 dynes/cm^2 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.
