Protocol for Robust In Vivo Measurements of Erythrocyte Aggregation Using Ultrasound Spectroscopy

Erythrocyte aggregation is a non-specific marker of acute and chronic inflammation. Although it is usual to evaluate this phenomenon from blood samples analyzed in laboratory instruments, in vivo real-time assessment of aggregation is possible with spectral ultrasound techniques. However, variable blood flow can affect the interpretation of acoustic measures. Therefore, flow standardization is required. Two techniques of flow standardization were evaluated with porcine and equine blood samples in Couette flow. These techniques consisted in either stopping the flow or reducing it. Then, the sensibility and repeatability of the retained method were evaluated in 11 human volunteers. We observed that stopping the flow compromised interpretation and repeatability. Conversely, maintaining a low flow provided repeatable measures and could distinguish between normal and high extents of erythrocyte aggregation. Agreement was observed between in vivo and ex vivo measures of the phenomenon (R² = 82.7%, p value < 0.0001). These results support the feasibility of assessing in vivo erythrocyte aggregation in humans by quantitative ultrasound means.