| Optimization of Microplate Aggregometry Method and Screening of Molecules with Antiplatelet Potential |
| Paper ID : 1181-IGA |
| Authors |
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Basma Hadjkacem *1, Asma Haffouz2, Ikram Ben Amor3, Jalel Gargouri4, Ali Gargouri2 1Laboratory of Molecular Biotechnology of Eucaryotes, Center of Biotechnology of Sfax, University of Sfax, Tunisia
Present address: Department of Life Sciences, Faculty of Sciences of Gafsa, University of Gafsa, Tunisia 2Laboratory of Molecular Biotechnology of Eucaryotes, Center of Biotechnology of Sfax, University of Sfax, Tunisia 3ikbeam@yahoo.fr 4Laboratory of Hematology, Medical Faculty of Sfax. University of Sfax, Tunisia |
| Abstract |
| Background and Aim: Antiplatelet drugs are commonly used to prevent thrombotic events. However, despite the variety of available medications, many are associated with side effects, highlighting the need for safer and more effective alternatives. Developing new molecules requires extensive screening. This study focuses on optimizing the microplate aggregometry technique using the Varioskan and screening a series of compounds to assess their antiplatelet potential. Materials and Methods: To assess microplate aggregometry, we optimized the agonist concentration, temperature, and stirring speed using the Varioskan. The optimal conditions were then applied to screen a series of chemical compounds. The results were validated using the photometric aggregometry method, which remains the gold standard for assessing pro- or anti-aggregant effects. Results: The optimization of microplate aggregometry conditions using the Varioskan identified the following optimal parameters: an agonist concentration of 40 µM ADP and 1.5 mM of arachidonic acid, shaking at 1000 rpm, and room temperature. Using these conditions, we screened a series of chemical compounds and identified four molecules with aggregation inhibitory effects: esculetin, methyl gallate, naringenin, and trans-ferulic acid. Their inhibition percentages were 62%, 60%, 18.5%, and 47%, respectively, for the arachidonic acid pathway, and 21%, 3%, and 37% for the ADP-induced pathway, while trans-ferulic acid showed no antiaggregant activity in the latter. All results were validated using the photometric aggregometry technique. Conclusion: This study highlights microplate aggregometry as a reliable, reproducible, and efficient method for screening antiplatelet molecules, requiring less platelet-rich plasma and allowing high-throughput testing. Using this technique, we successfully identified molecules with antiplatelet activity. |
| Keywords |
| Microplate aggregometry, varioskan, molecule screening, antiaggregant potential |
| Status: Accepted |
