title: Coadministration of adipose-derived stem cells and control-released basic fibroblast growth factor facilitates angiogenesis in a murine ischemic hind limb model.
abstract: OBJECTIVE
  Adipose-derived stem cells (ASCs) have angiogenic potential owing to their differentiation into endothelial cells and their release of angiogenic growth factors to elicit paracrine effects. In addition, control-released basic fibroblast growth factor (bFGF) sustained with a gelatin hydrogel also supports effective angiogenesis. We sought to determine if coadministration of ASCs and control-released bFGF into murine ischemic limbs facilitates angiogenesis.
  
  
  METHODS
  Levels of growth factors in the conditioned media of ASCs cultured with or without control-released bFGF were measured by enzyme-linked immunosorbent assays. A murine ischemic hind limb model was generated and intramuscularly injected with the following: gelatin hydrogel (group 1), a high number of ASCs (group 2), control-released bFGF (group 3), a small number of ASCs and control-released bFGF (group 4), and a high number of ASCs and control-released bFGF (group 5). Macroscopic and microscopic vascular changes were evaluated until day 7 by laser Doppler perfusion imaging and histologic analyses, respectively.
  
  
  RESULTS
  Secretion of hepatocyte growth factor, vascular endothelial growth factor, and transforming growth factor-β1 was enhanced by control-released bFGF. Vascular improvement was achieved in groups 4 and 5 according to laser Doppler perfusion imaging. Hematoxylin and eosin staining and CD31 immunohistochemical staining demonstrated an increase in the vascular density, vessel diameter, and thickness of vessel walls in groups 4 and 5. Cells positively stained for CD146, α-smooth muscle actin, and transforming growth factor-β1 were observed around vessel walls in groups 4 and 5.
  
  
  CONCLUSIONS
  These findings suggest that coadministration of ASCs and control-released bFGF facilitates angiogenesis in terms of vessel maturation in a murine ischemic hind limb model.
authors.affiliation: Department of Plastic and Reconstructive Surgery, Juntendo University School of Medicine, Tokyo, Japan.
authors.forename: Hisako
authors.initials: H
authors.lastname: Horikoshi-Ishihara
authors.affiliation: Department of Plastic and Reconstructive Surgery, Juntendo University School of Medicine, Tokyo, Japan.
authors.forename: Morikuni
authors.initials: M
authors.lastname: Tobita
authors.affiliation: Department of Plastic and Reconstructive Surgery, Juntendo University School of Medicine, Tokyo, Japan.
authors.forename: Satoshi
authors.initials: S
authors.lastname: Tajima
authors.affiliation: Department of Plastic and Reconstructive Surgery, Juntendo University School of Medicine, Tokyo, Japan.
authors.forename: Rica
authors.initials: R
authors.lastname: Tanaka
authors.affiliation: Department of Plastic and Reconstructive Surgery, Juntendo University School of Medicine, Tokyo, Japan.
authors.forename: Takashi
authors.initials: T
authors.lastname: Oshita
authors.affiliation: Department of Biomaterials, Institute for Frontier Medical Sciences, Kyoto University, Kyoto, Japan.
authors.forename: Yasuhiko
authors.initials: Y
authors.lastname: Tabata
authors.affiliation: Department of Plastic and Reconstructive Surgery, Juntendo University School of Medicine, Tokyo, Japan. Electronic address: hmizuno@juntendo.ac.jp.
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Mizuno
mesh_terms: D000273:Adipose Tissue / Q000166:cytology*; D043925:Angiogenesis Inducing Agents / Q000008:administration & dosage* / Q000737:chemistry; D000818:Animals; D002478:Cells, Cultured; D003131:Combined Modality Therapy; D017077:Culture Media, Conditioned / Q000378:metabolism; D003692:Delayed-Action Preparations; D004195:Disease Models, Animal; D004337:Drug Carriers; D016222:Fibroblast Growth Factor 2 / Q000008:administration & dosage* / Q000737:chemistry; D005780:Gelatin / Q000737:chemistry; D049452:Green Fluorescent Proteins / Q000096:biosynthesis / Q000235:genetics; D017228:Hepatocyte Growth Factor / Q000378:metabolism; D006614:Hindlimb; D006801:Humans; D020100:Hydrogels; D007511:Ischemia / Q000378:metabolism / Q000503:physiopathology / Q000628:therapy*; D008297:Male; D008810:Mice, Inbred C57BL; D008822:Mice, Transgenic; D018482:Muscle, Skeletal / Q000098:blood supply*; D018919:Neovascularization, Physiologic / Q000187:drug effects*; D020127:Recovery of Function; D012039:Regional Blood Flow; D033581:Stem Cell Transplantation; D013997:Time Factors; D053773:Transforming Growth Factor beta1 / Q000378:metabolism; D042461:Vascular Endothelial Growth Factor A / Q000378:metabolism
chemical_list: D043925:Angiogenesis Inducing Agents; D017077:Culture Media, Conditioned; D003692:Delayed-Action Preparations; D004337:Drug Carriers; C553307:HGF protein, mouse; D020100:Hydrogels; C508961:Tgfb1 protein, mouse; D053773:Transforming Growth Factor beta1; D042461:Vascular Endothelial Growth Factor A; C467480:vascular endothelial growth factor A, mouse; D016222:Fibroblast Growth Factor 2; D049452:Green Fluorescent Proteins; D017228:Hepatocyte Growth Factor; D005780:Gelatin