Study On Influence Of Angiogenesis Via Netrin-1 Allied With Mesenchymal Stem Cells In Arteriosclerosis Obliterans Rats

Background Peripheral arterial disease (PAD) is an atherosclerotic process that causes stenosis and occlusion of non-cerebral and non-coronary arteries. It has an estimated worldwide prevalence of almost 10%, rising to 15%～20% in people aged over 70. Arteriosclerosis obliterans (ASO) is a severe result from atherosclerosis. The gold-standard treatment of severe PAD is surgical or endovascular revascularization, however, up to 30% of patients are not candidates for such interventions, due to excessive operative risk or unfavorable vascular involvement. Although autologous mesenchymal stem cells (MSCs) transplantation is now proved to be an effective and safe method, there are still some defects that need to overcome. In recent years, many studies have demonstrated that netrin-1 has the ability of promoting angiogenesis besides the functions inthe nervous system, it could contribute to vascular remodeling as well. But the mechanism still remains unclear.Methods In vivo tests:Thirty-six rats were made into chronic limb ischemia models. They were randomly assigned to four groups, netrin-1+MSCs group (treated with netrin-1 and MSCs derived from peripheral blood), MSCs group (treated with MSCs individually), netrin-1 group (treated with netrin-1 individually), and control group (treated with saline). Measurements of murine behaviors, vascular endothelial growth factor expression, and capillary density in ischemia limb were performed on days 7,14, and 28 after treatments with autologous MSCs; measurements of contraction force in ischemia limb was performed on day 28 after treatments to compare differences among the groups. In vitro tests:Six rats were used for MSCs culture, then the MSCs were randomly assigned to four groups by different concentration of netrin-1 for cell migration assay and tube formation assay.Results Netrin-1 allied with MSCs significantly increased Tarlov score, vascular endothelial growth factor expression, capillary density, and muscular strength in ischemia limb. The higher concentration of netrin-1 resulted in more cells migrated and more tubes formed.Conclusions Netrin-1 allied with MSCs derived from peripheral blood significantly promoted angiogenesis in aged rats with chronic limb ischemia in vivo and in vitro. It may be a promising method of treating peripheral arterial disease in the future.