Figure 2

Extracorporeal perfusion preserves tissue integrity and reduces apoptotic damage;

(a) Histological images of rectus abdominis muscle tissue stained with H&E, 200 fold magnification, scale is 100 μm; 6 h, perfused: newly formed intercellular space visible as indicated exemplary by the black arrow, interpreted as interstitial edema; (b) Histological images of rectus abdominis muscle tissue stained against Annexin V, 200 fold magnification, scale is 100 μm; (c) Enlarged histological image of rectus abdominis muscle tissue stained with Annexin V, only nuclei within the border of the plasma membrane of skeletal muscle cells are considered myonuclei (as indicated by black arrows). Nuclei close to capillaries as well as outside of the plasma membrane have been excluded in further analysis to reduce unspecific staining and false positives, including endothelial cells and e.g. infiltrating neutrophils (as indicated by red arrows); (d) Ratio of Annexin V stained cell nuclei in relation to the total number of cell nuclei; Ratios of positive cell nuclei in relation to total number of cell nuclei for samples taken immediately after harvest (0 hours) before ischemia and after six hours of ischemia (6 hours), immediately after harvest (0 hours) before perfusion and after six hours of perfusion (6 hours). Mean ratios after six hours of ischemia and after six hours of perfusion rise statistically significant (*) compared to their individual values of 0 hours; (e) Ratio of Annexin V stained cell nuclei in relation to the total number of cell nuclei; mean difference of ratio changes of individual muscles undergoing six hours of ischemia or perfusion, with a statistically significant (*) lower rise in Annexin V positive cell nuclei of muscles undergoing perfusion. Error bars represent standard deviation of individual values recorded at the same time.