Low energy shock wave therapy induces angiogenesis in acute hind-limb ischemia via VEGF receptor 2 phosphorylation.

Johannes Holfeld,Marion Dietl,Daniela Lener,Daniela Lobenwein,Markus Theurl,Rudolf Kirchmair,Can Tepeköylü,Radoslaw Kozaryn,Michael Grimm,Patrick Paulus,Stefan Blunder,Elke Kirchmair

doi:10.1371/journal.pone.0103982

Abstract

ObjectivesLow energy shock waves have been shown to induce angiogenesis, improve left ventricular ejection fraction and decrease angina symptoms in patients suffering from chronic ischemic heart disease. Whether there is as well an effect in acute ischemia was not yet investigated.MethodsHind-limb ischemia was induced in 10–12 weeks old male C57/Bl6 wild-type mice by excision of the left femoral artery. Animals were randomly divided in a treatment group (SWT, 300 shock waves at 0.1 mJ/mm2, 5 Hz) and untreated controls (CTR), n = 10 per group. The treatment group received shock wave therapy immediately after surgery.ResultsHigher gene expression and protein levels of angiogenic factors VEGF-A and PlGF, as well as their receptors Flt-1 and KDR have been found. This resulted in significantly more vessels per high-power field in SWT compared to controls. Improvement of blood perfusion in treatment animals was confirmed by laser Doppler perfusion imaging. Receptor tyrosine kinase profiler revealed significant phosphorylation of VEGF receptor 2 as an underlying mechanism of action. The effect of VEGF signaling was abolished upon incubation with a VEGFR2 inhibitor indicating that the effect is indeed VEGFR 2 dependent.ConclusionsLow energy shock wave treatment induces angiogenesis in acute ischemia via VEGF receptor 2 stimulation and shows the same promising effects as known from chronic myocardial ischemia. It may therefore develop as an adjunct to the treatment armentarium of acute muscle ischemia in limbs and myocardium.

Highlights

Peripheral and coronary vascular disease still represent a major socio-economic health burden in industrialized countries
PlGF amplifies the angiogenic activity of vascular endothelial growth factor (VEGF) and induces further VEGF release
PlGF attracts smooth muscle cells for the coverage of capillaries and forms stable mature arterioles that are crucial for the proposed long-term benefit after shock wave treatment

Summary

Introduction

Peripheral and coronary vascular disease still represent a major socio-economic health burden in industrialized countries. The effect of surgical or interventional revascularization is limited due to nonviable myocytes. Clinicians are still in need of regenerative therapies as an adjunct to their treatment armentarium. Current treatment strategies for regeneration of infarcted muscle include (stem) cell or gene therapy based approaches. None of them yet gained broad clinical use due to distinct limitations [1]. Shown beneficial in numerous pre-clinical and clinical trials some challenges remain. The ideal cell type, the way of cell administration and homing of cells are still a matter of research in stem cell therapy [2,3,4]. Gene therapy with different kind of vectors as well shows promising results, delivery approaches and expression regulation of therapeutic gene products remain challenging [5,34]

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