Short-term pretreatment with low-dose hydrogen peroxide enhances the efficacy of bone marrow cells for therapeutic angiogenesis.
Therapeutic angiogenesis can be induced by the implantation of bone marrow cells (BMCs). Hydrogen peroxide (H(2)O(2)) has been shown to increase VEGF expression and to be involved in angiogenesis. We tested the hypothesis that pretreatment with H(2)O(2) enhances the efficacy of BMCs for neovascularization. H(2)O(2) pretreatment was done by incubating mouse BMCs in 5 microM H(2)O(2) for 30 min, followed by washing twice with PBS. The H(2)O(2)-pretreated and untreated BMCs were then studied in vitro and in vivo. RT-PCR analysis showed that expression of VEGF and Flk-1 mRNA was significantly higher in H(2)O(2)-pretreated BMCs than in untreated BMCs after 12 and 24 h of culture (P<0.01). Pretreatment with H(2)O(2) also effectively enhanced the VEGF production and endothelial differentiation from BMCs after 1 and 7 days of culture (P<0.05). To estimate the angiogenic potency in vivo, H(2)O(2)-pretreated or untreated BMCs were intramuscularly implanted into the ischemic hindlimbs of mice. After 14 days of treatment, many of the H(2)O(2)-pretreated BMCs were viable, showed endothelial differentiation, and were incorporated in microvessels. Conversely, the survival and incorporation of the untreated BMCs were relatively poor. Microvessel density and blood flow in the ischemic hindlimbs were significantly greater in the mice implanted with H(2)O(2)-pretreated BMCs than in those implanted with untreated BMCs (P<0.05). These results show that the short-term pretreatment of BMCs with low-dose H(2)O(2) is a novel, simple, and feasible method of enhancing their angiogenic potency.
Kubo
Masayuki
M
Department of Surgery and Clinical Science, Yamaguchi University, Graduate School of Medicine, Yamaguchi, Japan.
Li
Tao-Sheng
TS
Suzuki
Ryo
R
Ohshima
Mako
M
Qin
Shu-Lan
SL
Hamano
Kimikazu
K
D000818:Animals; D001854:Bone Marrow Cells / Q000187:drug effects* / Q000378:metabolism; D016026:Bone Marrow Transplantation; D002454:Cell Differentiation / Q000187:drug effects*; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D004195:Disease Models, Animal; D004305:Dose-Response Relationship, Drug; D042783:Endothelial Cells / Q000187:drug effects* / Q000378:metabolism; D005240:Feasibility Studies; D015870:Gene Expression / Q000187:drug effects; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D006614:Hindlimb / Q000098:blood supply; D006861:Hydrogen Peroxide / Q000494:pharmacology*; D007511:Ischemia / Q000378:metabolism / Q000503:physiopathology / Q000601:surgery*; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D008822:Mice, Transgenic; D008833:Microcirculation; D018919:Neovascularization, Physiologic / Q000187:drug effects*; D016877:Oxidants / Q000494:pharmacology*; D012333:RNA, Messenger / Q000378:metabolism; D012039:Regional Blood Flow; D013997:Time Factors; D042461:Vascular Endothelial Growth Factor A / Q000235:genetics / Q000378:metabolism; D040301:Vascular Endothelial Growth Factor Receptor-2 / Q000235:genetics / Q000378:metabolism
D016877:Oxidants; D012333:RNA, Messenger; D042461:Vascular Endothelial Growth Factor A; C485184:enhanced green fluorescent protein; C467480:vascular endothelial growth factor A, mouse; D049452:Green Fluorescent Proteins; D006861:Hydrogen Peroxide; D040301:Vascular Endothelial Growth Factor Receptor-2