Augmentation of revascularization by implantation of autologous bone marrow mononuclear cells in a rat ischemic hind limb model

Objective: To examine whether the in vivo implantation of autologous bone-marrow mononuclear cells (BM-MNCs) can augment postnatal neovascularization in rat model of hind limb ischemia.

Methods: Rat BM-MNCs were isolated by centrifugation through a Ficoll-paque density gradient. The rat ischemic hind limb model was made by ligation of the right femoral artery and its branches of imbred Wistar rats. BM-MNCs (MNC group) or phosphate saline buffer (PBS group) were injected into 7 points of the ischemic muscles (20 microl/point). To assess angiogenesis, histologic evaluation was performed on tissue slices from adductor muscles with immunohistochemical staining at the 2nd and 4th weeks after the ligation. Severity of ischemic insult was evaluated by the femoral arteriovenous oxgen difference (AVDO2) at the 2nd and 4th weeks after the ligation.

Results: The MNC groups had a higher capillary density and higher capillary/muscle fiber ratios than that of the sham and PBS groups (P < 0.01) at the 2nd and 4th weeks after the ligation. AVDO2 of the MNC group significantly decreased compared with that of the PBS groups (P < 0.05).

Conclusions: The implantation of autologous BM-MNCs induced neovascularization in a rat ischemic hind limb model. The implantation of autologous BM-MNCs augmented the collateral perfusion of ischemic hind limb.