In vitro evaluation of antithrombotic and anticoagulant properties of Silymarin-Silver Nanoparticles using human blood samples

Abstract

This study investigates the anticoagulant and antithrombotic properties of Silymarin Silver Nanoparticles (Sily-Ag NPs) using in vitro human blood samples. Silymarin, a natural flavonoid, was utilized for silver nanoparticles green synthesis, where it served as a reducing as well as a stabilizing agent. Comprehensive characterization of Sily-Ag NPs was done using UV-Vis spectroscopy, Fourier Transform Infrared spectroscopy (FTIR), X-ray Diffraction (XRD), Atomic Force Microscopy (AFM), Zeta potential analysis and Scanning Electron Microscopy (SEM). These analyses confirmed the nanoscale size, crystalline structure, stable surface charge, and effective capping by silymarin, suggesting the potential of these nanoparticles in biomedical applications. The study further evaluated the properties of antithrombosis and anticoagulation of Sily-Ag NPs using a series of coagulation assays, including Activated Partial Thromboplastin Time (aPTT), Fibrinogen levels, Prothrombin Time (PT), International Normalization Ratio (INR), and Thrombin time (TT) assessment. The results demonstrated that Sily-Ag NPs significantly prolonged PT, APTT, and INR, indicating a strong anticoagulant effect by interfering with both pathways of the coagulation cascade be it intrinsic or extrinsic. The findings also suggested that Sily-Ag NPs could effectively inhibit thrombin generation, a critical enzyme in the coagulation process. Interestingly, the Thrombin Time assay did not show any significant change, indicating that the nanoparticles do not directly interfere with concluding step of clot formation, the transformation of fibrinogen to fibrin. However, a modest reduction in fibrinogen levels was seen, which aligns with the overall anticoagulant and antithrombotic effects. The study concludes that Silymarin Silver Nanoparticles possess significant anticoagulant and antithrombotic properties, making them promising candidates for the development of new anticoagulant therapies. These nanoparticles demonstrated the ability to modulate multiple steps in the coagulation pathways without affecting the final fibrinogen-fibrin conversion, potentially offering a targeted therapeutic approach with fewer side effects compared to conventional anticoagulants. To determine Sily-Ag NPs' safety and effectiveness in clinical settings as well as to fully explore their therapeutic potential, more investigation is required, including in vivo experiments and clinical trials.