Article Figures & Data
- Figure 1
(A, B) The length of the silica tubing was adjusted to 1.5 mm using a digital calliper. (C) Schematic diagram of the intraoperative view. (D) Intraoperative picture under the microscope (see online supplemental movie 1). (E) Schematic diagram of the Evans blue injection centred on the anterior horn of the spinal cord (SC). (F) Representative example of Evans blue administration (0.7 µL) at angle X. (G) Overall image of the ventral side of the SC injected with 0.7 µL of Evans blue at each angle. (H) Diffusion of the dye in the SC, with 1.3 and 0.7 µL Evans blue. (I) Diffusion of the dye in the ventral SC was observed for two different volumes: 1.3 µL and 0.7 µL.
- Figure 2
Changes in the BBB locomotor score and body weight ratio. (A) The volume of ET-1 was fixed at 0.7 µL/side, and the concentration was varied. Each rat was observed for 14 days, and mortality rates are shown below: all five rats/group administered 1.0 mg/mL and 3.0 mg/mL ET-1, respectively, survived for 14 days; 1 of the 5 rats administered 2.0 mg/mL ET-1 died on day 4, and the other rats survived for 14 days. Data for the rat that died during the observation period were excluded from the graph. (B) ET-1 (0.7 µL/side, 2.5 mg/mL) was injected. (C) The BBB score on postoperative day 1 predicted the BBB score in the chronic phase. At 2 hours postoperatively, both the severe and mild groups had a BBB score of 0. Already on day 1 after the injection, however, the BBB score differed significantly, with the severe group having a mean BBB score of 0.26±0.10 and the Mild group having a BBB score of 2.33±0.36 (p<0.01). (D) The body weight changes in each group up to day 56. The average preoperative weight was defined as 1.0. BBB, Basso, Beattie and Bresnahan; PBS, phosphate-buffered saline.
- Figure 3
(A) TTC staining. Left: intact spinal cord (SC) at the Th13 level. Right: punctured cross-section on day 7n in the ET-1 group. (B) Vasoconstriction process after ET-1 injection. Blood flow at the level of Th13 was 61.6±7.4 mL/min/100 g before ET-1 administration, 15.9±4.0 mL/min/100 g immediately after ET-1 administration, 15.3±4.3 mL/min/100 g at 15 min, 31.9±1.4 mL/min/100 g at 30 min and 55.3±9.0 mL/min/100 g at 60 min. The blood flow was significantly decreased immediately, 15 min and 30 min after ET-1 administration compared with that before ET-1 administration (before vs just after; p<0.001, before vs 15 min; p<0.001, before vs 30 min; p<0.05). After 60 min, blood flow improved to the same level as before ET-1 administration (before vs 60 min; not significant). (C) Immunostaining with neurological markers, NeuN, Iba-1, GFAP and GST-pi. (D) Number of NeuN positive cells 3 days after ET-1 injection compared with intact SC. TTC, triphenyl tetrazolium chloride.
- Figure 4
(A) TUNEL-positive cells per unit area (mm2) 3 days after ET-1 injection. (B) Double staining with TUNEL and NeuN. (C) Immunostaining for RECA-1. (D) RECA-1 positive vessels per unit area (mm2). The number of RECA-1 positive vessels (/mm2) in the anterior horn was 428.9±26.9 and 457.8±4.6 (p=0.38) and the number in the posterior horn was 505.6±17.6 and 503.3±23.1 (p=0.93) in the ET-1- (day 3) and intact groups, respectively. In the white matter, the number of RECA-1-positive vessels (/mm2) was 192.2±9.2 and 172.2±8.2 (p=0.11) on the ventral side and 188.9±10.0 and 163.3±6.6 (p=0.04) on the dorsal side in the ET-1- (day 3) and intact groups, respectively.
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