Go Kuwahara1, Hitomi Nishinakamura2, Tadashi Tashiro1, Shohta Kodama2
1Department of Cardiovascular Surgery, Faculty of Medicine, Fukuoka University, Fukuoka City, Japan; 2Department of Regenerative Medicine & Transplantation, Faculty of Medicine, Fukuoka University, Fukuoka City, Japan.
OBJECTIVES: Bone marrow cells (BMCs) that include stem and progenitor cells are widely accepted to apply to patients suffering with peripheral arterial disease (PAD) as a therapeutic approach to induce neo-vascularization. However, the critical paracrine effects are still unclear to administrate BMCs. We focused macrophages that regulate VEGF, especially in VEGF-C.
METHODS: Male C57BL/6 mice were administered intramuscular injections of PBS control, unselected BMCs, CD11b+, CD11b- cells from BMCs, and recombinant VEGF-C to ischemic model of hind limb. As the evaluations, perfusion was measured with a laser Doppler scanning that were collected on days 0, 1, 3, 7, 14, 21, and 28 including a functional assay such as catwalk in parallel. Capillary density was determined by direct counting vessels positive with Von Willebrand factor at individual time points. Moreover, lymphangiogenesis was assessed as LYVE-1 positive cells.
RESULTS: Post-ischemic recovery of hind limb perfusion was significantly improved in BMCs, CD11b+ and VEGF-C group to compare with control group as the result of laser Doppler scanning. In the functional assay, VEGF-C group dramatically recovered compared to control group. The capillary/myofiber ratio in the thigh muscle was higher in BMCs and VEGF-C group than in control group same as LYVE-1 positive cells. Furthermore, the expression of VEGF, VEGF-C, their receptors of mRNA and Protein were involved predominately in CD11b+cells.
CONCLUSIONS: CD11b+macrophages play the critical role for angiogenesis and lymphangiogenesis in the murine model of hind limb ischemia. Consequently, recombinant VEGF-C accelerates the recovery and may be a promising therapeutic strategy for PAD patients.
AUTHOR DISCLOSURES: S. Kodama, Nothing to disclose; G. Kuwahara, Nothing to disclose; H. Nishinakamura, Nothing to disclose; T. Tashiro, Nothing to disclose.
Posted April 2012