Characterization of GPVI- or GPVI-CD39-Coated Nanoparticles and Their Impact on In Vitro Thrombus Formation |
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| Authors: | Jeremy A. Nestele,Anne-Katrin Rohlfing,Valerie Dicenta,Alexander Bild,Daniela Eiß ler,Frederic Emschermann,Marcel Kremser,Konstantin Krutzke,Tilman E. Schä ffer,Oliver Borst,Moran Levi,Netanel Korin,Meinrad Paul Gawaz |
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| Affiliation: | 1.Department of Cardiology and Angiology, University of Tübingen, 72076 Tubingen, Germany; (J.A.N.); (A.-K.R.); (V.D.); (A.B.); (D.E.); (F.E.); (M.K.); (O.B.);2.DFG Heisenberg Group Thrombocardiology, University of Tübingen, 72076 Tubingen, Germany;3.Institute for Applied Physics, University of Tübingen, 72076 Tubingen, Germany; (K.K.); (T.E.S.);4.Department of Biomedical Engineering, Technion, Israel Institute of Technology, Haifa 3200003, Israel; (M.L.); (N.K.) |
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| Abstract: | Traditional antithrombotic agents commonly share a therapy-limiting side effect, as they increase the overall systemic bleeding risk. A novel approach for targeted antithrombotic therapy is nanoparticles. In other therapeutic fields, nanoparticles have enabled site-specific delivery with low levels of toxicity and side effects. Here, we paired nanotechnology with an established dimeric glycoprotein VI-Fc (GPVI-Fc) and a GPVI-CD39 fusion protein, thereby combining site-specific delivery and new antithrombotic drugs. Poly(lactic-co-glycolic acid) (PLGA) nanoparticles, NP-BSA, NP-GPVI and NP-GPVI-CD39 were characterized through electron microscopy, atomic force measurements and flow cytometry. Light transmission aggregometry enabled analysis of platelet aggregation. Thrombus formation was observed through flow chamber experiments. NP-GPVI and NP-GPVI-CD39 displayed a characteristic surface coating pattern. Fluorescence properties were identical amongst all samples. NP-GPVI and NP-GPVI-CD39 significantly impaired platelet aggregation. Thrombus formation was significantly impaired by NP-GPVI and was particularly impaired by NP-GPVI-CD39. The receptor-coated nanoparticles NP-GPVI and the bifunctional molecule NP-GPVI-CD39 demonstrated significant inhibition of in vitro thrombus formation. Consequently, the nanoparticle-mediated antithrombotic effect of GPVI-Fc, as well as GPVI-CD39, and an additive impact of CD39 was confirmed. In conclusion, NP-GPVI and NP-GPVI-CD39 may serve as a promising foundation for a novel therapeutic approach regarding targeted antithrombotic therapy. |
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| Keywords: | nanoparticles hemostasis antithrombotic therapy platelets |
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