G-CSF and HGF: combination of vasculogenesis and angiogenesis synergistically improves recovery in murine hind limb ischemia.
Granulocyte colony-stimulating factor (G-CSF) is known to mobilize bone marrow stem cells into the peripheral circulation. This study was designed to investigate whether G-CSF by itself or in combination with hepatocyte growth factor (HGF) can promote vasculogenesis and angiogenesis in murine hind limb ischemia. Hind limb ischemia was induced in BALB/c nude or C57/BL6 mice that received bone marrow transplantation from green fluorescent protein (GFP)-transgenic mice. In the HGF group, hHGF expression plasmid was injected into the ischemic muscles. In the G-CSF group, G-CSF was administered subcutaneously for 10 days. The G-CSF+HGF group was concomitantly treated with G-CSF and HGF, and the control group received no treatment. All effects were confirmed at 4 weeks. The G-CSF+HGF group had a higher laser Doppler blood perfusion index, higher microvessel density, and a lower incidence of hind limb necrosis than the other groups. Confocal laser microscopy revealed that a number of GFP-positive cells infiltrated to the vasculature of the ischemic area. Some of the GFP positive cells were clearly co-immunostained with alpha-smooth muscle actin as well as von Willebrand factor. G-CSF-mobilized stem cells co-expressed CD49d and CD34, which would have promoted their adhesion to cells in the ischemic muscle that expressed HGF-induced vascular cell adhesion molecule-1. The combination of G-CSF and HGF had a significant synergistic effect, suggesting that the combination of mobilization of stem cells from bone marrow to peripheral circulation and their recruitment to the ischemic area might potentiate angiogenesis and vasculogenesis.
Ieda
Yasuyo
Y
Department of Regenerative Medicine and Advanced Cardiac Therapeutics, Keio University School of Medicine, 35 Shinanomachi, Shinjuku-ku, Tokyo 160-8582, Japan.
Fujita
Jun
J
Ieda
Masaki
M
Yagi
Takashi
T
Kawada
Hiroshi
H
Ando
Kiyoshi
K
Fukuda
Keiichi
K
D000818:Animals; D018952:Antigens, CD34 / Q000378:metabolism; D015815:Cell Adhesion Molecules / Q000378:metabolism; D004195:Disease Models, Animal; D004359:Drug Therapy, Combination; D017930:Genes, Reporter / Q000235:genetics; D016179:Granulocyte Colony-Stimulating Factor / Q000627:therapeutic use*; D017228:Hepatocyte Growth Factor / Q000627:therapeutic use*; D007511:Ischemia / Q000188:drug therapy* / Q000235:genetics / Q000378:metabolism / Q000473:pathology*; D017078:Laser-Doppler Flowmetry; D035002:Lower Extremity / Q000098:blood supply* / Q000473:pathology*; D008297:Male; D051379:Mice; D008819:Mice, Nude; D009336:Necrosis / Q000188:drug therapy / Q000473:pathology
D018952:Antigens, CD34; D015815:Cell Adhesion Molecules; D016179:Granulocyte Colony-Stimulating Factor; D017228:Hepatocyte Growth Factor