Development of reproducible histologic injury severity scores: Skeletal muscle reperfusion injury

Ischemic Preconditioning of Skeletal Muscle Mitigates Remote Injury and Mortality

“…There are a myriad of experimental ways to evaluate outcomes in I/R injury, including measures of histological evaluation, mitochondrial markers, and enzymatic activity [29]. We recently described a new statistical method of quantification through histology and used this method of evaluation in the current study [24].…”

“…Injured fibers displayed broken or ragged borders, a heterogeneous texture, the presence of intracellular holes, and/or detached nuclei. An injury score was obtained for each muscle by dividing the number of injured fibers by the total number of fibers for a given muscle, once a statistically significant number of fibers had been evaluated according to previously described criteria (600 myocytes per sample) [24].…”

Therapeutic delivery of hydrogen sulfide for salvage of ischemic skeletal muscle after the onset of critical ischemia

“…25 Determination of injury score. The injury score (IS) was determined by a protocol based on that which has been established by McCormack et al 26 Four random photomicrographs were taken of each H&E-stained section, and based on the proportion of injured cells (defined by ragged cellular borders, vacuolization, lymphocytic infiltration, or rhabdomyolysis), blinded reviewers assigned a numerical value between 0 and 10 for each in vivo experiment. Values are presented as mean Ϯ standard error of the mean.…”

Enoxaparin Does Not Ameliorate Limb Ischemia-Reperfusion Injury

“…Successful experimental therapies for skeletal muscle ischemia reperfusion have reliably demonstrated significant decrease in mediators of inflammation and decreased muscle fiber injury [32,36]. The methods used to analyze skeletal muscle injury in these experiments has provided reproducible quantitative analysis of muscle tissue injury [34], but there is no consensus in the literature regarding the best technique to quantitate muscle injury following reperfusion [37].…”

“…A random number generator was used to create a random series of image numbers, which a blinded observer used to retrieve the respective images and count muscle fibers. A total of 1200 muscle fibers per gastrocnemius sample (i.e., limb) were counted and scored as uninjured or injured based on the morphology of the individual fibers [34]. Uninjured fibers were characterized as having well-defined borders, consistent texture, and morphological uniformity without holes or breaks, with easily identifiable satellite cells and pericellular nuclei.…”