Regenerative effects of mouse aortic endothelial cells in a murine model of critical limb ischemia

Critical limb ischemia of the is a serious disease that threatens a significant decrease in working ability and disability of patients. Cell therapy may be useful in correcting the endothelial dysfunction that accompanies this disorder. The aim of study was to evaluate the effectiveness of local transplantation of mouse aortic endothelial cells (MAECs) in a model of critical limb ischemia in mice. Materials and methods. Critical limb ischemia in FVB mice was modeled by femoral artery ligation. The primary culture of endothelial cells was obtained from the murine aortic intima. The endothelial phenotype of cells for the expression of CD31, CD38 and CD309 markers was confirmed by flow cytometry and 1•106 MAECs were transplanted intramuscularly into ischemic limb. Tissue perfusion was assessed by laser Doppler flowmetry as well as descriptive histology was used to analyze changes in ischemic muscle after cell transplantation compared to the control group. Results. After MAECs transplantation in animals with modeled critical limb ischemia, the skin of the foot kept pink color and the corresponding temperature of the healthy limb without signs of necrosis of the distal phalanges in contrast to animals of the control group. According to laser Doppler flowmetry data, a significant difference (p ≤ 0.05) in perfusion of ischemic and sham-operated limbs in animals of the control group remained at the level of Δ = 45.7 ± 13.1 %. In animals after MAECs transplantation, the difference of these indicators between limbs was only Δ = 14.0 ± 8.23 % and was not statistically significant. A histological examination of muscle tissue after MAECs transplantation demonstrated the signs of compensatory processes characterized by hyperplasia and hypertrophy of myocyte’s nuclei and lightening of the nucleoplasm with well-defined nucleoli in some myofibrils. In the cytoplasm of myocytes, intermediate Z-discs were clearly visualized, and the number of myofibrils in muscle fibers increased. Conclusion. In animals with model of critical limb ischemia, the transplantation of aorta-derived endothelial cells recover the perfusion of ischemic limbs and improve the histological indicators of muscle tissue.