Abstract
Aims: Unsatisfactory effects of therapeutic angiogenesis in critical limb ischemia may be ascribed to use of circulating angiogenic cells (CACs) with impaired neovascularization-related capacities of atherosclerotic patients. We tested whether ultrasound cell stimulation can restore the impaired capacities. Methods and Results: During four-day-culture of human peripheral blood-derived mononuclear cells to achieve CACs, we stimulated the culture cells daily with low-intensity pulsed ultrasound stimulation (LIPUS). LIPUS to healthy volunteer-derived culture cells augmented generation and migration capacities of CACs, increased concentrations of angiopoietin 2 and nitrogen oxides in the culture medium, and increased expressions of phosphorylated-Akt and endothelial nitric oxide synthetase in CACs on western blotting. LIPUS to atherosclerotic patient-derived culture cells also augmented the generation and migration capacities of CACs. Although neovascularization in the ischemic hindlimb of athymic nude mice was impaired after intramuscular injection of atherosclerotic patient-derived CACs compared with that of healthy volunteer-derived CACs, LIPUS to atherosclerotic patient-derived culture cells restored the impaired neovascularization. Conclusion: Therapeutic angiogenesis with LIPUS-pretreated CACs may be a new strategy to rescue critical limb ischemia in atherosclerotic patients.
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