The Rat Ear Vein Model for Investigating In Vivo Thrombogenicity of Ultrafine Particles (UFP)

Abstract

Recent studies in rodents indicate that intravenous or intratracheal administration of ultrafine particles (UFP) increases thrombogenesis in a surgically exposed peripheral vein after photodynamic excitation of intravenously injected rose bengal (RB). We sought to adapt the invasive peripheral vein RB model to a noninvasive monitoring of ear veins under an inverted microscope. Animals received one of the following: an intraperitoneal, intravenous bolus, or intravenously infused dose of RB. An ear vein was illuminated by a green laser, and formation of a thrombus was captured with a digital camera. Only continuous intravenous infusion produced a steady-state RB plasma level and reproducible thrombus responses in different ear veins of the same rat. This system was then used to study the thrombogenic effects of iv-administered positively or negatively charged 60-nm ultrafine polystyrene particles (PSP). Significant dose-dependent enhancement of thrombus formation was found, as indicated by decreased laser illumination time to 33% of baseline values at 0.5 mg/kg. Negatively charged PSP of the same size failed to affect thrombus formation. We also studied the thrombogenic effect of PSP without the use of RB. The findings were the same as with RB, although the illumination time had to be increased. When 0.5 mg/kg was instilled intratracheally, the laser illumination time to form a thrombus was decreased to 42% of the baseline value, suggesting translocation of UFP into the bloodstream. These results are consistent with previous findings using the invasive model, and they validate the use of this non-invasive ear vein model to evaluate thrombogenic effects of UFP deposition in the respiratory tract.