Formación de trombos inducida por láser en el Endo-chip: un modelo de vaso en chip para pruebas in vitro de agentes antitrombóticos

Mouse models, such as the intravital laser-induced model of thrombus formation, are commonly used for mechanistic and preclinical studies in thrombosis. However, their translational value is limited by species differences. Few in vitro models incorporate laser-induced vascular injury. Here, we developed an endothelialized microfluidic device, the Endo-chip, to model thrombus formation in response to laser injury. Citrated blood was treated with IXA4, an endoplasmic reticulum stress inducer, or with antithrombotic agents including antitissue factor antibody (5G9), antiplatelet drugs (abciximab, aspirin), and anticoagulants (argatroban, heparin). Fluorescently labeled antibodies (to platelets, fibrin, or von Willebrand factor [VWF]), annexin V or the calcium dye Cal520, were added to the blood. After recalcification at 10 mM, blood was perfused through the Endo-chip at a shear rate of 100 s-1 or 800 s-1. Endothelial injury was induced with a 355-nm laser pulse producing a focal 10-μm injury, and phosphatidylserine exposure on endothelial cells within ∼1 cell diameter from the injury site. Annexin V-positive endothelial cells expressed tissue factor and released VWF, supporting localized platelet and fibrin deposition. The thrombus formed a teardrop morphology aligned with flow incorporating VWF with increasing shear. IXA4 enhanced platelet cytoplasmic calcium. Platelet accumulation was inhibited by abciximab but not aspirin, whereas coagulation inhibitors (5G9, argatroban, heparin) markedly reduced thrombus formation. These findings support that the Endo-chip laser-injury model incorporates key features of thrombus formation after endothelial injury, and provides a humanized, in vitro, alternative or auxiliary to mouse models for preclinical studies and antithrombotic drug development.

Resumen tomado de PubMed (NCBI) para el registro citado, en el idioma original. La versión de referencia es la de la publicación original.

El resumen se deja sin traducir para conservar la redacción original de los autores.

Los autores desarrollaron el Endo-chip, un dispositivo microfluídico endotelizado, para modelar la trombogénesis tras lesión láser. Se perfundió sangre citratada a tasas de cizallamiento de 100 s-1 u 800 s-1; un pulso láser de 355 nm generó una lesión focal de 10 µm. Abciximab inhibió la acumulación plaquetaria, no así el ácido acetilsalicílico; 5G9, argatrobán y heparina redujeron marcadamente la trombogénesis. Según los autores, el modelo reproduce rasgos clave y constituye una alternativa humanizada in vitro a modelos murinos.

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