Mononuclear cell therapy reverts cuff-induced thrombosis in apolipoprotein E-deficient mice

Leandro C F Lima,Marcella L Porto,Silvana S Meyrelles,Breno V Nogueira,Clarissa L Tonini,Bianca P Campagnaro,Elisardo C Vasquez,Thiago Mc Pereira

doi:10.1186/1476-511x-11-96

Abstract

BackgroundStem/progenitor cell-based therapy has successfully been used as a novel therapeutic strategy for vascular diseases triggered by endothelial dysfunction. The aim of this study was to investigate the effects of mononuclear cell (MNC) therapy in situ on carotid cuff-induced occlusive thrombus in the apolipoprotein E knockout (apoE-/-) mouse.MethodsSpleen-derived MNCs were isolated from green fluorescent protein (GFP)-transgenic mice for cell treatment. A cuff-induced thrombus model was produced by placing a nonconstrictive silastic collar around the left common carotid artery in 20-week-old female apoE-/- mice. After 10 days, the cuff was removed, and the animals received in situ MNCs (Cuff-MNC) or vehicle (Cuff-Vehicle) and were compared with sham-operated animals (Sham).ResultsThe histological analysis showed that the MNC treatment reverted occlusive thrombus formation compared to the vehicle and the vessel lumen area to that observed in the Sham group (MNC, 50 ± 4; Vehicle, 20 ± 4; Sham, 55 ± 2 x103 μm2; p < 0.01). The animals that underwent the carotid cuff placement developed compensatory vessel enlargement, which was reduced by the MNC therapy. In addition, the treatment was able to reduce superoxide anion production, which likely contributed to the reduced apoptosis that was observed. Lastly, the immunofluorescence analysis revealed the presence of endothelial progenitor cells (EPCs) in the carotid endothelia of the apoE-/- mice.ConclusionIn situ short-term MNC therapy was able to revert cuff-induced occlusive thrombi in the carotid arteries of apoE-/- mice, possibly through the homing of EPCs, reduction of oxidative stress and decreased apoptosis.

Highlights

Stem/progenitor cell-based therapy has successfully been used as a novel therapeutic strategy for vascular diseases triggered by endothelial dysfunction
The apolipoprotein E-deficient mouse is considered to be the most relevant model because the animals develop spontaneous hypercholesterolemia and endothelial dysfunction accompanied by arterial lesions that are similar to those observed in humans [1,7]
Animals ApoE-/- female mice (20 weeks old) were randomly divided into three groups: an apoE-/- control group (Sham, n = 8), apoE-/- treated in situ with mononuclear cell (MNC) group (Cuff-MNC, n = 8) and apoE-/- treated with DMEM group (Cuff-Vehicle, n = 8)

Summary

Introduction

Stem/progenitor cell-based therapy has successfully been used as a novel therapeutic strategy for vascular diseases triggered by endothelial dysfunction. The development of ischemic diseases in conditions of hypercholesterolemia is aggravated by endothelial dysfunction [5,6]. The apolipoprotein E-deficient (apoE-/-) mouse is considered to be the most relevant model because the animals develop spontaneous hypercholesterolemia and endothelial dysfunction accompanied by arterial lesions that are similar to those observed in humans [1,7]. Recent studies have demonstrated that the female gender is an important factor in the development of endothelial dysfunction and atherosclerosis [1,8,9]. Because the integrity of the endothelium is important to suppress thrombosis, the cuff placement model may be an interesting strategy to study thrombogenesis by the impairment of arterial distensibility

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