Abstract / Summary
Objective Thromboembolic disorders remain a major global health concern, especially in the context of vascular injury and inflammation. In this study, we developed FITC-dextran-labeled gelatin nanoparticles (Gel-FITC NPs) conjugated with thrombin to simulate pulmonary thromboembolic conditions in vitro. Materials and Methods Gelatin was selected for its biocompatibility and ease of nanoparticle formation via the desolvation method, while FITC-dextran was incorporated to enable in vitro fluorescent detection and tracking. Thrombin was covalently attached to preserve enzymatic activity and enhance procoagulant functionality. The nanoparticles were characterized using DLS, SEM, UV–Vis, FTIR, and XRD, confirming successful synthesis and conjugation. Results After thrombin grafting, the average particle size changed to 320 ± 30 nm and the surface charge shifted to −19 mV. The grafting efficiency results indicated that a 1:1 thrombin to Gel-FITC NPs mass ratio offers the most efficient (80%) and reproducible conjugation conditions. Hemocompatibility was evaluated through hemolysis assays, and coagulation potential was assessed via PT, aPTT, and fibrin formation tests. Results demonstrated reduced clotting times and enhanced fibrin dynamics in the presence of thrombin-conjugated nanoparticles. Conclusion While in vivo biodistribution and therapeutic efficacy warrant future investigation, this dual-functional platform offers a promising in vitro tool for modeling thromboembolic events and screening candidate therapies under controlled laboratory conditions.