Figure 1.
Murine model of Laser-Doppler Flowmetry assisted acute ischemia-reperfusion in hindlimb.
(A) Anaesthetized mouse with ligated hindlimb. The vessel loop was fixed advancing a small, solid plastic tube with a mosquito clamp throughout the whole ischemia period. (B–C) Blood perfusion to the hindlimb was assessed using Laser-Doppler Flowmetry (moorVMS-LDF1) (a) before ischemia, (b) during ischemia and (c) directly when reperfusion is induced. Error bars represent Standard Error of the Mean (SEM).
Figure 2.
Tissue regeneration after acute ischemia and reperfusion.
Haematoxylin and eosin staining showed regeneration of (A) the skin and (B) muscle tissue after 4 hours of ischemia and at different times after reperfusion. Scale bar 200 µm.
Figure 3.
Analysis of angiogenic factors during tissue regeneration.
(A) In muscle, VEGF (green) is progressively up-regulated at 6 and 12 hours of reperfusion; 14 days after injury it had returned to base level. Scale bar 50 µm. (B) CD34-positive angiogenic stem cells (red) followed VEGF up-regulation and progressively infiltrated the affected areas starting after 96 hours of reperfusion. Scale bar 20 µm.
Figure 4.
Analysis of angiogenesis during tissue regeneration.
(A) Assessment of CD31-positive capillaries at different intervals of reperfusion after 4 h acute ischemia in skin (B) and muscle. *p<0.05. The mean values of CD31 positive capillaries are given in Table 1 and 2, respectively.
Table 1.
Assessment of CD31-positive capillaries in the skin (n = 6).
Table 2.
Assessment of CD31-positive capillaries in the muscle (n = 6).