Early protective effect of bone marrow mononuclear cells against ischemic white matter damage through augmentation of cerebral blood flow.
OBJECTIVE
To investigate the efficacy of bone marrow mononuclear cell (BMMNC) treatment against ischemic white matter (WM) damage in a hypoperfused brain.


METHODS
Mice were administered intravenous treatment of vehicle, spleen-derived marrow mononuclear cells (MNCs), or BMMNCs (5 × 10⁶ cells) obtained from enhanced green fluorescent protein transgenic mice 24 hours after bilateral common carotid artery stenosis (BCAS), and then euthanized at either 1 day or 30 days after treatment.


RESULTS
Laser speckle perfusion imaging analyses revealed marked recovery of cerebral blood flow (CBF) in the early phase after BMMNC treatment (6 hours after administration), before histological evidence of angiogenesis was assessed by fluorescein-isothiocyanate-dextran perfusion assay. BMMNC treatment induced an increase in vascular endothelial growth factor and Ser1177 phosphorylated endothelial nitric oxide synthase levels in the BCAS-induced mouse brains at 1 day after the treatment. BCAS-induced ischemic WM lesions were significantly improved 30 days after BMMNC treatment despite any evidence of direct structural incorporation of donor BMMNCs into endothelial cells and oligodendrocytes. Instead, enhanced green fluorescent protein-positive donor cells with morphological features of pericytes were observed in the vessel walls. Post-BMMNC administration of an NOS inhibitor abolished early CBF recovery and produced protective effects against ischemic WM damage.


CONCLUSIONS
BMMNC treatment provides marked protection against ischemic WM damage, enhancing CBF in the early phase and in subsequent angiogenesis, both of which involve nitric oxide synthase activation. These findings suggest promise for the application of BMMNCs for subcortical ischemic vascular dementia.
Fujita
Youshi
Y
Department of Neurology, Graduate School of Medicine, Kyoto University, Kyoto 606-8507, Japan. fujitauc@kuhp.kyoto-u.ac.jp
Ihara
Masafumi
M
Ushiki
Takashi
T
Hirai
Hideyo
H
Kizaka-Kondoh
Shinae
S
Hiraoka
Masahiro
M
Ito
Hidefumi
H
Takahashi
Ryosuke
R
D000818:Animals; D015153:Blotting, Western; D016026:Bone Marrow Transplantation; D001921:Brain / Q000473:pathology*; D001923:Brain Chemistry / Q000502:physiology; D002545:Brain Ischemia / Q000473:pathology / Q000628:therapy*; D002196:Capillaries / Q000473:pathology; D016893:Carotid Stenosis / Q000473:pathology / Q000628:therapy; D002454:Cell Differentiation / Q000502:physiology; D002560:Cerebrovascular Circulation / Q000502:physiology*; D004797:Enzyme-Linked Immunosorbent Assay; D005434:Flow Cytometry; D005455:Fluorescent Antibody Technique; D049452:Green Fluorescent Proteins; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D008822:Mice, Transgenic; D018919:Neovascularization, Physiologic / Q000502:physiology; D052250:Nitric Oxide Synthase Type III / Q000096:biosynthesis; D010766:Phosphorylation; D042461:Vascular Endothelial Growth Factor A / Q000378:metabolism
D042461:Vascular Endothelial Growth Factor A; D049452:Green Fluorescent Proteins; D052250:Nitric Oxide Synthase Type III; C496315:Nos3 protein, mouse