Modified multipotent stromal cells with epidermal growth factor restore vasculogenesis and blood flow in ischemic hind-limb of type II diabetic mice

Abstract

Diabetes is increasing in the world and causes severe cardiovascular complications. Diabetes-induced limb ischemia leads to foot amputation and therapeutic remedies are urgently needed. Here we report that local injection of mesenchymal stem cells (MSCs) prestimulated with epidermal growth factor (EGF) restored blood flow and vasculogenesis in the ischemic hind-limb of type II diabetic (db−/db−) mice. Bone marrow cells from db−/db− mice are altered as evidenced by increased oxidative stress and reduced Akt and adhesion molecules when compared with control (db−/db+). Femoral artery ligation-induced ischemia was performed in the hind-limb of db−/db− and db−/db+ mice for 28 days. Enhanced green fluorescent protein (EGFP)-MSCs stimulated±exogenous EGF for 24 h were injected locally into the ischemic muscle. Blood flow measured with MoorLDI-Laser and microangiography assessed with X-ray showed 100% recovery in db−/db+ compared to 50% recovery in db−/db− mice. Interestingly, db−/db− mice had 60 and 96% blood flow recovery and 61 and 98% of vasculogenesis when treated with MSCs alone or MSCs modified with EGF, respectively. Western blot analysis of hind-limb muscles revealed an increase in Akt and vascular endothelial growth factor receptor phosphorylation and hypoxia-inducible factor) expression in db−/db− mice injected with MSCs or MSCs +EGF compared to db−/db− mice. Fluorescent microscopic images show that EGFP-MSCs differentiate into new microvessels. Adhesion and migration of MSCs on cultured endothelial cells were ICAM1-, VCAM1- and Akt-dependent mechanism and elevated when MSCs were prestimulated with EGF compared with nonstimulated MSCs. Our novel study data provide evidence that in type II diabetes, stimulated MSCs with EGF enhance the recovery of blood flow and angiogenesis.

Coronary vein infusion of multipotent stromal cells from bone marrow preserves cardiac function in swine ischemic cardiomyopathy via enhanced neovascularization

“…Yet, the cell implantation failed to reduce cardiac fibrosis or achieve reverse remodeling in this study. Further studies in large animals to explore the methods to enhance or modify MSC function, such as gene transfer 11 and pharmacological preconditioning, 31 will be necessary.…”

Vasodilator responses to acetylcholine are not mediated by the activation of soluble guanylate cyclase or TRPV4 channels in the rat

“…After 1 h incubation, the PA rings were frozen in liquid nitrogen. The cGMP measurements were performed using an enzyme-linked immunosorbent assay kit from Cayman Chemical (Ann Arbor, MI) according to the manufacturer's protocol and as previously described (1,6).…”

Osteoporosis in diabetes mellitus: Possible cellular and molecular mechanisms

“…Xu and coworkers (2009) demonstrated that injection of BMSC treated with pancreatic extract into streptozotocinindu ced DM rats not only normalized plasma glucose and pre vented apoptosis of islet cells, but also elevated produc tion of VEGF, IGF-1 and basic fibroblast growth factor (bFGF), all of which are known to have antiapoptotic and angiogenic effects [66] . A recent in vivo study in type 2 DM (db -/db -) mice with ischemic hind limbs showed that injection of epidermal growth factor (EGF)treated BMSC into the affected hind limbs increased angiogene sis by over 90% [67] . Such angiogenesis was due to the fact that the injected BMSC differentiated into new mi crovessels (neovascularization), using intercellular adhe sion molecule1 and vascular cell adhesion protein1 for adhesion and migration [67] .Overall, it is possible that anti diabetic agents with angiogenic activity could be used to enhance blood flow to fracture sites, which may in turn accelerate bone healing, and might also prevent osteo penia/osteoporosis.…”

“…A recent in vivo study in type 2 DM (db -/db -) mice with ischemic hind limbs showed that injection of epidermal growth factor (EGF)treated BMSC into the affected hind limbs increased angiogene sis by over 90% [67] . Such angiogenesis was due to the fact that the injected BMSC differentiated into new mi crovessels (neovascularization), using intercellular adhe sion molecule1 and vascular cell adhesion protein1 for adhesion and migration [67] .Overall, it is possible that anti diabetic agents with angiogenic activity could be used to enhance blood flow to fracture sites, which may in turn accelerate bone healing, and might also prevent osteo penia/osteoporosis. Conversely, certain rheological drugs, such as pentoxifylline, which increase blood flow and osteoblast activity, might be promising as antiosteoporotic agents in both DM and nonDM patients [68] .…”