Changes in B-mode Ultrasound Echo Intensity Following Injection of Bupivacaine Hydrochloride to Rat Hind Limb Muscles in Relation to Histologic Changes

This study tested the hypothesis that infiltration of inflammatory cells in muscle fibers would increase echo intensity (image brightness) of B-mode ultrasound images. Bupivacaine hydrochloride (BPVC) or saline solution (SAL) was injected to the tibialis anterior (TA) muscles of 14- to 23-wk-old male Wistar rats. Ultrasound images were taken from the muscles before and at 0, 2, 4, 6, 9, 12, 24, 48, 72, 120, 168 and 336 h after the injection and analyzed for the echo intensity (echogenicity) expressed as the mean value of image pixel value of a region-of-interest. Changes in the echo intensity were compared between BPVC-injected and control or SAL-injected muscles. In the subsequent study, rats (n = 2 per time point) were sacrificed after taking ultrasound image at 0, 2, 6, 12, 24, 48 and 168 h after BPVC injection to the right TA and SAL injection to the left TA to observe histologic changes under a light microscope and the relationship between echo intensity and inflammatory cells was assessed. No significant changes in echo intensity were observed for the control, but BPVC induced significant (p < 0.05) increases in the echo intensity peaking 0 to 24 h postinjection. SAL also increased echo intensity immediately after injection but returned to the baseline by 24 h postinjection. The time course of changes in the echo intensity did not match with the time course of increases in inflammatory cells in the muscle. It is concluded that infiltration of inflammatory cells is not a direct cause of the increased echo intensity.