26 January 2018 : Animal Research
The Role of miR-126 in Critical Limb Ischemia Treatment Using Adipose-Derived Stem Cell Therapeutic Factor Concentrate and Extracellular Matrix MicroparticlesVáclav Procházka1ABCDEFG*, Jana Jurčíková2ABDEFG, Kateřina Vítková2ABDEFG, Lubomír Pavliska2CD, Ludmila Porubová3ABDEFG, Ondrej Lassák3CD, Piotr Buszman4BDEF, Carlos A. Fernandez4ABDE, František Jalůvka5AB, Iveta Špačková6BDE, Ivo Lochman7BDE, Martin Procházka8ADEF, Mária Janíková89ADEF, Zdeněk Tauber10ADEF, Jana Franková11ADEF, Martin Lachnit2BDE, Michael C. Hiles12DE, Brian H. Johnstone13ADEF
Med Sci Monit 2018; 24: ANS511-522
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.
Keywords: Culture Media, Conditioned, In Situ Hybridization, stem cell research
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