BACKGROUND Paracrine factors secreted by adipose-derived stem cells can be captured, fractionated, and concentrated to produce therapeutic factor concentrate (TFC). The present study examined whether TFC effects could be enhanced by combining TFC with a biological matrix to provide sustained release of factors in the target region. MATERIAL AND METHODS Unilateral hind limb ischemia was induced in rabbits. Ischemic limbs were injected with either placebo control, TFC, micronized small intestinal submucosa tissue (SIS), or TFC absorbed to SIS. Blood flow in both limbs was assessed with laser Doppler perfusion imaging. Tissues harvested at Day 48 were assessed immunohistochemically for vessel density; in situ hybridization and quantitative real-time PCR were employed to determine miR-126 expression. RESULTS LDP ratios were significantly elevated, compared to placebo control, on day 28 in all treatment groups (p=0.0816, p=0.0543, p=0.0639, for groups 2-4, respectively) and on day 36 in the TFC group (p=0.0866). This effect correlated with capillary density in the SIS and TFC+SIS groups (p=0.0093 and p=0.0054, respectively, compared to placebo). A correlation was observed between miR-126 levels and LDP levels at 48 days in SIS and TFC+SIS groups. CONCLUSIONS A single bolus administration of TFC and SIS had early, transient effects on reperfusion and promotion of ischemia repair. The effects were not additive. We also discovered that TFC modulated miR-126 levels that were expressed in cell types other than endothelial cells. These data suggested that TFC, alone or in combination with SIS, may be a potent therapy for patients with CLI that are at risk of amputation.

American Heart of Poland Inc Katowice Poland

Cook Biotech Inc West Lafayette IN USA

Department of Clinical and Molecular Pathology Palacky University Olomouc and University Hospital Olomouc Olomouc Czech Republic

Department of Deputy Director of Science and Research University Hospital Ostrava Ostrava Czech Republic

Department of Histology and Embryology Faculty of Medicine and Dentistry Palacky University Olomouc Olomouc Czech Republic

Department of Medical Chemistry and Biochemistry Faculty of Medicine and Dentistry Palacky University Olomouc Olomouc Czech Republic

Department of Medical Genetics University Hospital Olomouc and Palacky University Olomouc Olomouc Czech Republic

Department of Surgery University Hospital Ostrava Ostrava Czech Republic

Laboratoře AGEL a s Nový Jičín Czech Republic

MEDi CBTD Ostrava Czech Republic

NeuroFX LLC Indianapolis IN USA

Radiodiagnostic Institute University Hospital Ostrava Ostrava Czech Republic

SPADIA LAB a s Ostrava Czech Republic

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Current Status of Cell-Based Therapy in Patients with Critical Limb Ischemia