title: Therapeutic Potential of Adipose-Derived Therapeutic Factor Concentrate for Treating Critical Limb Ischemia.
abstract: Transplantation of adipose-derived stem cells (ADSCs) is an emerging therapeutic option for addressing intractable diseases such as critical limb ischemia (CLI). Evidence suggests that therapeutic effects of ADSCs are primarily mediated through paracrine mechanisms rather than transdifferentiation. These secreted factors can be captured in conditioned medium (CM) and concentrated to prepare a therapeutic factor concentrate (TFC) composed of a cocktail of beneficial growth factors and cytokines that individually and in combination demonstrate disease-modifying effects. The ability of a TFC to promote reperfusion in a rabbit model of CLI was evaluated. A total of 27 adult female rabbits underwent surgery to induce ischemia in the left hindlimb. An additional five rabbits served as sham controls. One week after surgery, the ischemic limbs received intramuscular injections of either (1) placebo (control medium), (2) a low dose of TFC, or (3) a high dose of TFC. Limb perfusion was serially assessed with a Doppler probe. Blood samples were analyzed for growth factors and cytokines. Tissue was harvested postmortem on day 35 and assessed for capillary density by immunohistochemistry. At 1 month after treatment, tissue perfusion in ischemic limbs treated with a high dose of TFC was almost double (p < 0.05) that of the placebo group [58.8 ± 23 relative perfusion units (RPU) vs. 30.7 ± 13.6 RPU; mean ± SD]. This effect was correlated with greater capillary density in the affected tissues and with transiently higher serum levels of the angiogenic and prosurvival factors vascular endothelial growth factor (VEGF) and hepatocyte growth factor (HGF). The conclusions from this study are that a single bolus administration of TFC demonstrated robust effects for promoting tissue reperfusion in a rabbit model of CLI and that a possible mechanism of revascularization was promotion of angiogenesis by TFC. Results of this study demonstrate that TFC represents a potent therapeutic cocktail for patients with CLI, many of whom are at risk for amputation of the affected limb.
authors.affiliation: Radiodiagnostic Institute, University Hospital Ostrava, Ostrava, Czech Republic.
authors.forename: Václav
authors.initials: V
authors.lastname: Procházka
authors.forename: Jana
authors.initials: J
authors.lastname: Jurčíková
authors.forename: Ondrej
authors.initials: O
authors.lastname: Laššák
authors.forename: Kateřina
authors.initials: K
authors.lastname: Vítková
authors.forename: Lubomír
authors.initials: L
authors.lastname: Pavliska
authors.forename: Ludmila
authors.initials: L
authors.lastname: Porubová
authors.forename: Piotr P
authors.initials: PP
authors.lastname: Buszman
authors.forename: Agata
authors.initials: A
authors.lastname: Krauze
authors.forename: Carlos
authors.initials: C
authors.lastname: Fernandez
authors.forename: František
authors.initials: F
authors.lastname: Jalůvka
authors.forename: Iveta
authors.initials: I
authors.lastname: Špačková
authors.forename: Ivo
authors.initials: I
authors.lastname: Lochman
authors.forename: Dvořáčková
authors.initials: D
authors.lastname: Jana
authors.forename: Stephanie
authors.initials: S
authors.lastname: Merfeld-Clauss
authors.forename: Keith L
authors.initials: KL
authors.lastname: March
authors.forename: Dmitry O
authors.initials: DO
authors.lastname: Traktuev
authors.forename: Brian H
authors.initials: BH
authors.lastname: Johnstone
mesh_terms: D000273:Adipose Tissue / Q000378:metabolism*; D000818:Animals; D002478:Cells, Cultured; D017077:Culture Media, Conditioned / Q000494:pharmacology; D016207:Cytokines / Q000627:therapeutic use; D005260:Female; D005434:Flow Cytometry; D017228:Hepatocyte Growth Factor / Q000378:metabolism; D006614:Hindlimb / Q000473:pathology*; D006801:Humans; D007273:Injections, Intramuscular; D036341:Intercellular Signaling Peptides and Proteins / Q000627:therapeutic use; D007511:Ischemia / Q000188:drug therapy*; D059630:Mesenchymal Stem Cells / Q000378:metabolism; D018919:Neovascularization, Physiologic / Q000187:drug effects; D011817:Rabbits; D042461:Vascular Endothelial Growth Factor A / Q000378:metabolism
chemical_list: D017077:Culture Media, Conditioned; D016207:Cytokines; D036341:Intercellular Signaling Peptides and Proteins; D042461:Vascular Endothelial Growth Factor A; D017228:Hepatocyte Growth Factor