Intrathecal transplantation of bone marrow stromal cells attenuates blood-spinal cord barrier disruption induced by spinal cord ischemia-reperfusion injury.

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Intrathecal transplantation of bone marrow stromal cells attenuates blood-spinal cord barrier disruption induced by spinal cord ischemia-reperfusion injury in rabbits Bo Fang, MD, He Wang, MD, Xue-Jun Sun, MD, Xiao-Qian Li, MD, Chun-Yu Ai, MD, PhD, Wen-Fei Tan, MD, PhD, Paul F. White, PhD, MD, Hong Ma, MD, PhD Journal of Vascular Surgery Volume 58, Issue 4, Pages 1043-1052 (October 2013) DOI: 10.1016/j.jvs.2012.11.087 Copyright © 2013 Society for Vascular Surgery Terms and Conditions

Fig 1 Effects of bone marrow stromal cells (BMSCs) on neurologic motor function and histologic assessment of the spinal cord after spinal cord ischemia-reperfusion (IR) injury. A, Tarlov score at 4 and 24 hours after injury (n = 12). B, Number of intact motor neurons in the ventral gray matter. ∗∗P < .01 vs group sham; ##P < .01 vs group IR. Data are presented as mean ± standard error (n = 6). C, Representative sections of lumbar spinal cords stained with hematoxylin and eosin (original magnification, ×100). Journal of Vascular Surgery 2013 58, 1043-1052DOI: (10.1016/j.jvs.2012.11.087) Copyright © 2013 Society for Vascular Surgery Terms and Conditions

Fig 2 Effects of bone marrow stromal cells (BMSCs) on Evans blue (EB) extravasation after spinal cord ischemia-reperfusion (IR) injury. Almost no red fluorescence was seen in spinal cord parenchyma in (A) group sham and (B) group sham + BMSCs. C, Much red fluorescence could be seen at 4 hours after injury. E, This turned stronger at 24 hours after injury, especially in gray matter. D and F, In contrast, EB red fluorescence was significantly weakened in group IR + BMSCs. G, Quantification data of EB content of spinal cord (μg/g) are presented as mean ± standard error (n = 6). ∗∗P < .01 vs group sham; ##P < .01 vs group IR. Journal of Vascular Surgery 2013 58, 1043-1052DOI: (10.1016/j.jvs.2012.11.087) Copyright © 2013 Society for Vascular Surgery Terms and Conditions

Fig 3 Effects of bone marrow stromal cells (BMSCs) on the ultrastructure of the blood-spinal cord barrier by transmission electron microscopy at 24 hours after spinal cord ischemia-reperfusion (IR) injury. A, In group sham, the lanthanum nitrate was exclusively localized in the capillary. B, In group IR, a leakage of lanthanum was found in the basement membrane (BM), even invading into the parenchyma along the tight junction (TJ). C, In group IR + BMSCs, only little extravasation of lanthanum was observed. The unlabeled arrowheads indicate the leakage of lanthanum. L, Capillary lumen. Scale bar: 1 μm. Journal of Vascular Surgery 2013 58, 1043-1052DOI: (10.1016/j.jvs.2012.11.087) Copyright © 2013 Society for Vascular Surgery Terms and Conditions

Fig 4 Bone marrow stromal cell (BMSC) transplantation suppressed the decreases in the tight junction protein occludin after spinal cord ischemia-reperfusion (IR) injury. A, Representative Western blot and quantitative protein analysis of occludin in the spinal cord at 4 and 24 hours after injury. B, Real-time polymerase chain reaction analysis shows occludin messenger RNA (mRNA) expression. The expression level of each sample is expressed as the ratio to group sham. ∗∗P < .01 vs group sham; ##P < .01 vs group IR. Data are presented as mean ± standard error (n = 6). C, Immunofluorescence photomicrographs of occludin (green) along microvasculature (red) 24 hours after injury. Journal of Vascular Surgery 2013 58, 1043-1052DOI: (10.1016/j.jvs.2012.11.087) Copyright © 2013 Society for Vascular Surgery Terms and Conditions

Fig 5 Transplantation of bone marrow stromal cells (BMSCs) suppressed the upregulation of matrix metalloproteinase (MMP)-9 and tumor necrosis factor (TNF)-α after spinal cord ischemia-reperfusion (IR) injury. A, Representative Western blot and quantitative protein analysis of MMP-9 in spinal cord at 4 and 24 hours after injury. B, Real-time polymerase chain reaction analysis of MMP-9 messenger RNA (mRNA) expression. C, Representative Western blot and quantitative protein analysis of TNF-α in spinal cord at 4 and 24 hours after injury. The expression level of each sample is expressed as the ratio to group sham. ∗∗P < .01 vs group sham; ##P < .01 vs group IR. Data are presented as mean ± standard error (n = 6). Journal of Vascular Surgery 2013 58, 1043-1052DOI: (10.1016/j.jvs.2012.11.087) Copyright © 2013 Society for Vascular Surgery Terms and Conditions