Beneficial effects of concurrent autologous bone marrow cell therapy and metabolic intervention in ischemia-induced angiogenesis in the mouse hindlimb

Abstract

Lower-limb ischemia is a major health problem. Because of the absence of effective treatment in the advanced stages of the disease, amputation is undertaken to alleviate unbearable symptoms. Novel therapeutic approaches include the intramuscular use of autologous bone marrow cells (BMCs). Because tissue ischemia is associated with an overwhelming generation of oxygen radicals and negative effects due to perturbed shear-stress, metabolic intervention with antioxidants and L-arginine could potentially induce beneficial effects beyond those achieved by BMCs. The protective effect of autologous BMCs and vascular protection by metabolic cotreatment (1.0% vitamin E added to the chow and 0.05% vitamin C and 6% L-arginine added to the drinking water) were examined in ischemia-induced angiogenesis in the mouse hindlimb, a model of extensive acute peripheral arterial occlusion. i.v. BMC therapy improved blood flow and increased capillary densities and expression of Ki-67, a proliferation-associated protein. This beneficial effect was amplified by metabolic cotreatment, an intervention inducing vascular protection, at least in part, through the nitric oxide pathway, reduction of systemic oxidative stress, and macrophage activation. Therefore, although a cautious approach is mandatory when experimental findings are extended to human diseases, autologous BMCs together with metabolic intervention could be an effective clinical treatment for peripheral arterial disease.antioxidants ͉ ischemic hindlimb ͉ L-arginine ͉ peripheral arterial disease ͉ nitric oxide

Brain protection using autologous bone marrow cell, metalloproteinase inhibitors, and metabolic treatment in cerebral ischemia

“…The fresh brain slices were immersed for 30 min in a 2% solution of TTC (57). The cross-sectional area of infarction in each brain slice was examined with a dissection microscope and measured by using imaging analysis software (9,25,26). For measurements in the cortex, eight fields of view from the cortical ischemic penumbra (cortical IBZ) were digitized.…”

“…BMCs were obtained autologously from the femurs of rats 72 h before transplantation, as described (9). The same rats from which BMCs were obtained (groups 2-4) were subjected later to MCAO.…”

“…Sections were counterstained with hematoxylin. Quantification of BrdU-immunoreactive cells was performed on paraffin-embedded tissue sections and counted digitally (9,25,26,59). To identify expression of cell type-specific markers in BrdU ϩ cells, double immunofluorescence was performed (9,59).…”

“…To date, virus-mediated overexpression of TIMPs has not been used as a means to confer neuroprotection in vivo. Furthermore, neuroprotection could be improved by metabolic treatment, a concept supported by a great body of evidence indicating beneficial roles of treatment with antioxidants and L-arginine, the natural precursor of NO, during vascular dysfunction (9,(25)(26)(27)(28). Antioxidants mitigate oxidative and neuronal damage after ischemic stroke (29)(30)(31).…”

“…Bone marrow cells (BMCs) contain populations of precursors that are multipotent and can differentiate into bone, cartilage, and mesenchyma (7), neurons and glia (8), and endothelial cells (9,10). BMCs have been demonstrated to cross the blood-brain barrier (BBB) (11).…”