Evaluation of the Sensitivity of an in vitro High Frequency Ultrasound Device to Monitor the Coagulation Process: Study of the Effects of Heparin Treatment in a Murine Model

Abstract 

This study evaluates the sensitivity of a new in vitro high frequency ultrasound test of the whole blood coagulation process. A rat model of anticoagulant treatment is reported. Many recent studies of the role of red blood cells in the whole blood coagulation process have revealed an increasing demand for global tests of the coagulation process performed on whole blood instead of plasma samples. In contrast to existing optical tests, high frequency ultrasound presents the advantages of characterizing the mechanical properties of whole blood clotting. Ultrasound longitudinal wave velocity and integrated attenuation coefficient (IAC) were simultaneously assessed in a 10 to 30MHz frequency range during the whole blood coagulation process in vitro in rats under anticoagulant therapy. Differences between humans and rats were also clearly emphasized in non-clotting blood and in clotting blood using specific criteria deduced from acoustic parameters (ultrasound velocity for non-clotting blood:=1574±2m/s for rats and 1583±3m/s for humans and IAC=2.25±0.14 dB/cm for rats and 1.5±0.23 dB/cm for humans). We also measured the coagulation time t₀ from the acoustic velocity (t₀ =11.15±7min for control rat blood and 43.3±11.4min for human blood). Different doses of heparin were administered to rats. The sensitivity of the ultrasound device to the effects of heparin was evaluated. Differences between non-treated rats and chronically and acutely treated rats were recorded and quantified. We particularly noted that the slope S and the amplitude I of the variations in acoustic velocity were linked to clot retraction, which is a good indicator of the platelet function. The amplitude of the variations in S was between (20±8)×10^–3 m/s² for control group rats, and (0.92±0.35)×10^–3 m/s² for chronic heparin-treated group rats. The values of I were 15 times higher for control group rats than for chronic heparin-treated group rats. (E-mail: rachel.libgot@univ-tours.fr)

Key Words: High frequency ultrasound, Blood clotting, Acoustic velocity, Anticoagulant therapy, Rat, In vitro