Ultrasonographic assessment of skeletal muscles after experimentally induced neurogenic inflammation (facet injury) in rats

Abstract

This study set out to examine ultrasonographic attributes of non-neurosegmentally (pectoral-forelimb) and neurosegmentally linked (hindlimb) myotomes in an experimental model that leads to neurogenic inflammation in segmentally linked myotomes, and to evaluate quantitative correlations among ultrasonographic attributes of the muscles, relative content of various inflammatory mediators, and nociceptive thresholds (hot and mechanical) in rats. Twelve male Wistar Kyoto rats were randomly divided into two equinumerous groups: surgery group, in which the left lumbar (L4–L6) facet joints were compressed for 3 min with modified Kelly forceps under general anesthesia, and sham-operated rats. All ultrasonograms were obtained with the Vevo 2100 Visual Sonic scanner connected to a 24-MHz transducer at four different time points: pre-surgery and 7, 14, and 21 days after surgical procedures. Digital ultrasonographic images of quadriceps femoris, hamstring, and pectoral-brachial muscle groups were analyzed using a polygonal meter region of interest placed on the largest cross-sectional area of the muscles displayed in Image ProPlus® analytical software to compute numerical pixel values and pixel heterogeneity (standard deviation of mean pixel values). On day 21, pain behavior tests (hot plate and von Frey) were performed and then all animals were euthanized. Protein expression of inflammatory mediators in biceps brachii and rectus femoris muscles was measured by Western blot. The most prominent differences in muscle echotextural attributes between the two subsets of rats occurred 14 days post-surgery in pectoral-brachial and quadriceps femoris muscles. The expression of calcitonin-gene-related peptide was directly related to both echotextural variables only in biceps brachii (pixel intensity: r = 0.65, P = 0.02; and heterogeneity: r = 0.66, P = 0.02, respectively). Our findings have revealed the occurrence of echotextural changes in skeletal muscles of rats during myositis; however, the accumulation of inflammatory mediators and the outcomes of sensory tests did not relate to the changes in first-order echotextural characteristics of affected hindlimb muscles.

Ultrasound-based assessment of the expression of inflammatory markers in the rectus femoris muscle of rats

“…The Animal Care Committee of the University of Guelph (Guelph, ON, Canada) had approved all experimental procedures involving live animals. The present retrospective study used the original data and ultrasound images obtained and analyzed by Duarte et al [14] and Ahmadi et al [15], but with the new discriminating image analysis of muscle ultrasonograms. Twelve sexually mature male Wistar Kyoto rats, aged 13 ± 5 months, were housed (2-3 animals per cage) in a room with a 12-hour alternating light-dark cycle and a stable temperature (23.0 ± 1.0 °C).…”

“…Transcutaneous ultrasonography of various muscle groups in the rats of the present sudy has been detailed previously [14,15]. All ultrasound images of the rectus femoris muscle 21 days after surgical interventions were recorded in B-mode using the Vevo 2100 Visuals Sonic Ultrasound Diagnostic Medical Imaging System (Visual Sonic Inc., Toronto, ON, Canada; Figure 2).…”

“…All animals were euthanized with carbon dioxide on Day 21 of the experiment (Day 0 = surgery) and the samples of the rectus femoris were collected for Western blot analyses of inflammatory regulators, as described previously by Ahmadi et al [15]. Muscle samples (each 20-30 mg) were frozen and then homogenized with cell lysis buffer (NP40 CLB-FNN0021; Thermo Scientific Fisher, Canada) containing protease inhibitor and serine protease inhibitor…”