Citation: HE Fei, ZHANG Tian-Cong, CHEN Dan, DING Ling, Deng Su-Rong, MI Yong-Hua, LIU Zhan-Shu, LI Yuan. Analysis of Dose-effect Relationship of Tirofiban Inhibiting Platelet Adhesion Aggregation Using Microscopic Three-dimensional Imaging Technology[J]. Chinese Journal of Analytical Chemistry, ;2022, 50(2): 244-252. doi: 10.19756/j.issn.0253-3820.210671 shu

Analysis of Dose-effect Relationship of Tirofiban Inhibiting Platelet Adhesion Aggregation Using Microscopic Three-dimensional Imaging Technology

HE Fei ¹ ,
ZHANG Tian-Cong ¹ ,
CHEN Dan ¹ ,
DING Ling ² ,
Deng Su-Rong ¹ ,
MI Yong-Hua ¹ ,
LIU Zhan-Shu ¹ ,
LI Yuan ^1,,

1.
Central Laboratory of Yongchuan Hospital, Chongqing Medical University, Yongchuan 402160, China;
2.
Yongchuan Sub-center of Chongqing Blood Center, Yongchuan 402160, China

Corresponding author: LI Yuan, liyuan_1985999@163.com
Received Date: 11 August 2021
Revised Date: 8 December 2021

Fund Project: Supported by the National Natural Science Foundation of China (No.11702047), the Special Project of Science and Technology Innovation for People′s Livelihood Security of Chongqing (No.cstc2017shmsA130009), the Chongqing Medical Research Program (No.2017MSXM079), the Special Project of Science and Technology Innovation for Livelihood Security of Yongchuan (No.Ycstc2018cc0206) and the Research Project of Yongchuan Hospital, Chongqing Medical University(No.YJLC202033)

The dose-effect relationship of tirofiban (a platelet membrane glycoprotein (GP) Ⅱb/Ⅲa receptor antagonist) in inhibiting platelet adhesion and aggregation was qualitatively and quantitatively evaluated using a novel microscopic three-dimensional (3D) imaging technology. The platelet-rich plasma (PRP) of healthy volunteers was firstly treated by 0-50 μg/mL tirofiban, and then added to collagen fibrin and glass surfaces respectively to induce platelet activation and aggregation. The 3D morphological images of platelet adhesion and aggregation under different experimental conditions were observed by laser microscopic 3D imaging technology. The volume and area coverage rate of platelet aggregates, and the number and activated count of adhesion platelet were quantified. The results showed that the 3D micro-morphology of platelet aggregates formed on the surfaces of collagen fibrin and glass could be visually observed by the laser microscopic 3D imaging technology. The platelet aggregates formed on the surface of collagen fibrin were thin, and those on the surface of glass were hill-like. When the final concentration of tirofiban was 1.56-12.5 μg/mL, the volume and area coverage rate of platelet aggregate decreased significantly with the increase of tirofiban final concentration. When the concentration increased to 25 and 50 μg/mL, platelets adhered to collagen fibrin only, and the number of platelet adhesion and the proportion of activated platelets decreased significantly with the increase of concentration. On the glass surface, the tirofiban with concentration of 1.56 μg/mL could reduce the volume and area coverage rate of platelet aggregates; and when the concentration was 3.15-50 μg/mL, it could completely inhibit platelet aggregation, and the number of platelet adhesion and the proportion of activated platelets decreased with the increase of tirofiban concentration. As a result, when the concentration was 3.15-12.5 μg/mL, tirofiban could not only completely inhibit platelet aggregation on the glass surface but retain partial aggregation ability on collagen fibrin surface. The microscopic 3D imaging technique could quantitatively and qualitatively analyze the dose-effect relationship of tirofiban inhibiting platelet adhesion aggregation on collagen fibrin surface and glass surface, which provided a new analytical method for rational clinical selection of tirofiban dose.
- Platelet aggregation,
- Microscopic three-dimensional morphology,
- Collagen fiber,
- Glass,
- Tirofiban,
- Does-effective relationship
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