Ultrasound stimulation restores impaired neovascularization-related capacities of human circulating angiogenic cells.
OBJECTIVE
Unsatisfactory effects of therapeutic angiogenesis in critical limb ischaemia may be ascribed to use of circulating angiogenic cells (CACs) derived from atherosclerotic patients with impaired neovascularization-related capacities. We tested whether ultrasound cell stimulation can restore the impaired capacities.


RESULTS
During culture of human peripheral blood-derived mononuclear cells for 4 days to achieve CACs, we stimulated the cells in culture daily with low-intensity pulsed ultrasound stimulation (LIPUS). Application of LIPUS to cells in culture derived from healthy volunteers augmented the generation and migration capacities of CACs, increased concentrations of angiopoietin 2 and nitrogen oxides in the culture medium, and increased the expression of phosphorylated-Akt and endothelial nitric oxide synthase in CACs on western blotting. Application of LIPUS to cells in culture derived from atherosclerotic patients also augmented the generation and migration capacities of CACs. Although neovascularization in the ischaemic hindlimb of athymic nude mice was impaired after intramuscular injection of CACs derived from atherosclerotic patients compared with that using CACs derived from healthy volunteers, LIPUS of the cells in culture derived from atherosclerotic patients restored the neovascularization capacities.


CONCLUSIONS
Therapeutic angiogenesis with LIPUS-pre-treated CACs may be a new strategy to rescue critical limb ischaemia in atherosclerotic patients.
Department of Internal Medicine, Division of Cardio-Vascular Medicine,  Kurume University School of Medicine, 67 Asahi-machi, Kurume 830-0011,  Japan.
Yasuyuki
Y
Toyama
Ken-ichiro
K
Sasaki
Katsuro
K
Tachibana
Takafumi
T
Ueno
Hidemi
H
Kajimoto
Shinji
S
Yokoyama
Masanori
M
Ohtsuka
Hiroshi
H
Koiwaya
Takaharu
T
Nakayoshi
Yoshiaki
Y
Mitsutake
Hidetoshi
H
Chibana
Naoki
N
Itaya
Tsutomu
T
Imaizumi
D042702:Angiopoietin-2 / Q000378:metabolism; D000818:Animals; D050197:Atherosclerosis / Q000378:metabolism / Q000473:pathology* / Q000503:physiopathology; D015153:Blotting, Western; D016022:Case-Control Studies; D002465:Cell Movement; D049109:Cell Proliferation; D002478:Cells, Cultured; D004195:Disease Models, Animal; D006614:Hindlimb; D006801:Humans; D007511:Ischemia / Q000378:metabolism / Q000473:pathology / Q000503:physiopathology / Q000628:therapy; D007963:Leukocytes, Mononuclear / Q000378:metabolism / Q000473:pathology* / Q000637:transplantation; D051379:Mice; D008807:Mice, Inbred BALB C; D008819:Mice, Nude; D018482:Muscle, Skeletal / Q000098:blood supply; D018919:Neovascularization, Physiologic; D009569:Nitric Oxide / Q000378:metabolism; D052250:Nitric Oxide Synthase Type III / Q000378:metabolism; D010641:Phenotype; D010766:Phosphorylation; D051057:Proto-Oncogene Proteins c-akt / Q000378:metabolism; D015398:Signal Transduction; D013997:Time Factors; D014465:Ultrasonics; D015854:Up-Regulation
D042702:Angiopoietin-2; D009569:Nitric Oxide; C496314:NOS3 protein, human; D052250:Nitric Oxide Synthase Type III; D051057:Proto-Oncogene Proteins c-akt