Application of a Simple Microfluidic Chip Analysis Technology to Evaluate the Inhibitory Role of Protocatechuic Acid on Shear-Induced Platelet Aggregation.

Haidong Ma,Mohammad Hashem Hashempur,Cui He,Wenran Dan,Yuan Li,Surong Deng,Lihua Yu,Beizhong Liu

doi:10.1155/2021/5574413

Abstract

This study aimed to develop a simple microfluidic chip analysis technology to study the inhibitory effect of protocatechuic acid on shear-induced platelet aggregation. The microfluidic chip designed in this study simulates 80% fixed narrow microchannels. This microchannel narrow model uses the finite element analysis module of the three-dimensional modeling software solidwork to analyze fluid dynamic behavior. Blood treated with protocatechuic acid at 1, 2, 4, 8, or 16 µg/mL was passed through the microchannel stenosis model at a shear rate of 10,000 s−1. The platelet adhesion and aggregation behaviors were then measured using fluorescence microscopy and observed in real time. Simultaneously, the antiplatelet aggregation effect of protocatechuic acid was analyzed using thromboelastography and photoelectric turbidimetry. The designed stenosis model of the microfluidic chip can produce a gradient of fluid shear rate, and the gradient of fluid shear rate can induce platelet aggregation. Under this model, the degree of platelet adhesion and aggregation increased as the shear rate increased. In the experimental concentration range of 0–8 µmol/mL, protocatechuic acid exerted a concentration-dependent inhibition of platelet aggregation. In contrast, thromboelastography and photoelectric turbidimetry failed to demonstrate an inhibitory effect. The microfluidic chip analysis technology developed in this study can be used to study the effect of protocatechin in inhibiting platelet aggregation induced by shear rate in vitro. This technology is simple to operate and can be used as a new type of antiplatelet aggregation analysis technology for screening studies of novel potential antiplatelet aggregation drugs.

Highlights

Adhesion and aggregation of platelets at the site of vascular injury are the basis of normal hemostasis and pathological thrombosis
In the state of thrombotic disease, mild to severe arterial stenosis can reach ∼2,000–8,000 s−1, and the maximum wall shear rate of severely affected atherosclerotic arteries can be as high as 40,000 s−1 [5, 6]. erefore, shear-induced platelet aggregation (SIPA) is a special type of platelet aggregation and the main cause of pathological thrombosis, and it is rarely seen in normal hemostasis
When the shear rate is low, the fluid velocity is slow moving away from the constriction. erefore, platelets will slowly aggregate at the place where the downstream flow slows, gradually forming stable platelet microaggregates. is has been demonstrated in our previous experiments. erefore, the platelet adhesion aggregation image studied in this study selected the aggregation area downstream of the stenosis at 0.75 mm

Summary

Introduction

Adhesion and aggregation of platelets at the site of vascular injury are the basis of normal hemostasis and pathological thrombosis. Inhibition of this function poses a risk of increased bleeding [1]. The shear rate of the vessel wall in the vein is approximately 10–200 s−1 and in the aorta is approximately 300–800 s−1. E wall shear rate of small arteries is approximately 450–1,600 s−1 [4]. In the state of thrombotic disease, mild to severe arterial stenosis can reach ∼2,000–8,000 s−1, and the maximum wall shear rate of severely affected atherosclerotic arteries can be as high as 40,000 s−1 [5, 6]. In the state of thrombotic disease, mild to severe arterial stenosis can reach ∼2,000–8,000 s−1, and the maximum wall shear rate of severely affected atherosclerotic arteries can be as high as 40,000 s−1 [5, 6]. erefore, shear-induced platelet aggregation (SIPA) is a special type of platelet aggregation and the main cause of pathological thrombosis, and it is rarely seen in normal hemostasis

Methods

Results

Conclusion