Combined therapy with DMOG-treated BMDACs and AdCA5 improves perfusion and clinical outcome after hindlimb ischemia in 17-month-old mice. (A) Laser Doppler perfusion imaging (top and middle panels) and photography (bottom panel) were performed at the indicated time after femoral artery ligation. The top panel shows typical scans of female 17-month-old mice immediately before (Pre) and after (Post) femoral artery ligation in the left hindlimb. Blood flow is quantified using a pseudocolor scale as shown at right (perfusion units [PU]). The middle panel shows representative follow-up images of mice treated with saline, AdCA5, or combined therapy (AdCA5 + BMDAC-D). Because saline- and AdCA5-treated mice had severe tissue necrosis, the last time point (day 14 [D14]) before autoamputation ensued is shown. Bottom panel: Representative photographs of mice at day 28 after surgery (D28). (B) Mean ischemic/nonischemic limb perfusion ratio, as determined by laser Doppler perfusion imaging, was calculated at the indicated time (in days) after hindlimb ischemia. Mice were treated with saline (black), intramuscular AdCA5 (blue), or intramuscular AdCA5 + intravenous BMDAC-D (green). Intramuscular AdCA5 was injected immediately after surgery and intravenous BMDAC-D were delivered 24 hours later. *P < .05 vs saline control by Bonferroni post hoc test after ANOVA (n = 3 mice per group). (C) Tissue damage (left panel) was scored as follows: 0 indicates no tissue damage; 1, cyanosis/discoloration indicative of soft tissue necrosis; 2, loss of 1 or 2 toes; 3, loss of 3 to 5 toes; 4, amputation of entire foot; 5, amputation of entire lower leg; and 6, amputation of entire leg. Motor impairment (right panel) was scored as follows: 0 indicates normal response (plantar/toe flexion in response to tail traction); 1, plantar but not toe flexion; 2, no plantar or toe flexion; and 3, dragging of foot. Motor impairment and ischemic tissue damage were measured 28 days after surgery. *P < .05 vs saline control by Bonferroni post hoc test after ANOVA (n = 3 mice per group). Data are mean ± SEM.