149 Angiogenesis Induced by Enriched Bone Marrow Cell Transplantation in Ischemic Hindlimb Canine Model

Bone marrow is an important supply for many progenitor cells, among them endothelial precursor cells. They have been already considered as potential therapeutic angiogenesis for ischemic hindlimb. In this work we performed an unilateral chronic ischemic hindlimb model in 20 dogs, we were dissected and ligated the middle sacra artery and external right iliac artery preserving the internal iliac artery, after one week we removed de femoral artery performing a proximal and distal ligation and inserted into the gracilis muscle 3 Silastic tubes (0.8 mm diameter) in a middle circle form in order to created fibrocollagenous tunnels. Fifteen days later, we obtained bone marrow (30 mL) and mononuclear cells were separated; at the same time, all tubes were removed and bone marrow cells were transplanted into the fibrocollagenous tunnels, only in the third and fourth groups. During the 8 at 12 days we administered subcutaneously saline solution to the first and third groups and G‐CSF to the second and fourth groups. Finally, after 30 days we performed contrasted angiographies from both limbs and was calculated mean angiographic score (MAS) from the number of vascular intersections, also was taken a biopsy from the central portion of right gracilis muscle for vascular assessment, as well as for vascular proliferating cells by immunohistochemistry. Results demonstrated that white bone marrow cell transplantation enrichment by G‐CSF administration stimulates angiogenesis in ischemic hindlimb, twice than controls, and it is better than white bone marrow cell transplantation alone. White bone marrow cell is a potential therapy for ischemic hindlimb, because stem cells are growth factor provider, as well as an important source for endothelial precursors capable to form vessels de novo , mainly when this stem cells are deposited in an appropriate extracellular matrix, such as the fibrocollagenous tunnels in this model.