The Animal Experimental Study Of Transplantation With Endothelial Progenitor Cells For Treating Arteriosclerosis Obliterans

Objeetive:To observe the treatment possibility and effects on rats models with experimental femoral artery obliterans by transplantation of endothelial progenitor cells(EPCs) from bone marrow mesenchymal stem cells(MSCs).Methods:Experimental rats models with both femoral arteries obliterans were successful made by dissecting,ligating,and cutting the branches of both femoral arteries which were clamped by forceps for 30s in 18 rats, as well as were fed high-fat diet. A11 animals were intramuscular injected 300 OOOu/kg vitamin D3 once per month. MSCs were isolated from allogeneic male wistar rats,then were cultured. VEGF,EGF,bFGF and IGF-1 were added into culture medium and the active endothelial progenitor cells were labled with BrdU.The cells were transplanted into right hindlimbs by intramusculer injection as group B in rats of experimental femoral obliterans models,and same volume of normal saline were injected into the opposite hindlimbs as group A as control. On 14 day and 28 day hindlimbs were extracted for immunohistochemical examinations(by FV111 and BrdU).Results:The numbers of blood vessels with positive staining of FactorVlll in hindlimb at 14d after transplantation in group B1 (4.556±0.506)were more than that in group Al (2.867±0.915) (P<0.05); The numbers of blood vessels with positive staining of FactorVlll in hindlimb at 28d after transplantation in group B2 (5.941±1.324)were obviously more than that in group A2 (3.000±0.707) (P<0.05) Some positive stained endothelial cells by BrdU were found in A but not in B.Conclusion:The arteriosclerosis obliterans models were established successfully.Transplantation of EPCs from cultured MSCs in vitro can significantly increase the capillary density in ischemic hindlimbs,in rats with experimental femoral artery obliterans as compared with that in control hindlimbs.The BrdU positive staining in capillary in group A may suggest that the EPCs which we transplanted into ischemic tissue can improve and increase the angiogenesis. The EPCs may come from bone marrow cells.They have potential to differentiate into vascular endothelial cells in ischemic tissue in vivo. The differentiation and development potentiality of EPCs to vasoendothelial cells in ischemic tissue was much better than that of naturally auto-angiogenesis in vivo.