To investigate the effects of fibrin glue on repair and regeneration of acute complete spinal cord injury. Methods Acute complete transaction spinal cord injury model were made in 10 adult healthy SD rats(female, weighing 250-300 g), randomized grouping: treated group(n=5) and control group(n=5). In the treated group, fibrin glue was implanted covering on the injury site and fill ing the lesion gap. In the control group, no treatment was given. At 4 weeks, the locomotor functions of the rats were detected by basso, beattie and bresnahan (BBB) score, then the means of immunohistochemistry were used to observe neurofilament(NF) and gl ial fibrillary acidic protein(GFAP). And image analysis was used to measure the quantify of the nerve fiber and the fibers area ratio of astrocyte. Results The BBB scores were 2.40 ± 0.51 in control group, 3.00 ± 0.45 in treated group, showing no significant difference (P gt; 0.05). By immunohistochemistry: a l ittle positive NF cells and GFAP frame were found in control group; more positive NF cells and GFAP frame were found in treated group, the cells and frame grew toward the center but did not arrive at the center. Image analysis showed the amount of never fibers in treated group (rostral region: 113.10 ± 20.75, caudal region: 73.60 ± 33.61) was more than that in control group (rostral region: 45.50 ± 17.18, caudal region 23.50 ± 8.20), showing significant difference. The fibers area ratio of astrocyte in treated group(rostral region: 33.75% ± 11.06%, caudal region: 27.75% ± 7.15%) was more than that in control group(rostral region: 23.78% ± 5.76%, caudal region: 19.78% ± 5.17%), showing significant difference (P lt; 0.05). Conclusion Fibrin glue can promote repair and regeneration of acute spinal cord injury.
Thirty-five SD rats were divided into 3 groups, in which 5 rats were served as control. The other 2 groups, 15 rats in each received either NGF solution or normal saline. The injury at the level of T8 spinal segment of the rat in these two groups were made by dropping a weight of 10 g from a height 2.5 cm after a total laminectomly from T7-11 segments. A thin plastic tube was inserted into the subarachnoid space below the injured segments. NGF was introduced through the tube at intervals of 1, 2, 3, 4, 8, 12, 24, 48 hours in the NGF group, and normal saline in the normal saline group. At 4, 8, 24 hours following surgery, 5 rats in each group were sacrificed and the injured segments were resected for examination. The contents of water and calcium were measured by dry-wet method and atomic absorption spectroscopy. The result showed that total calcium and water contents in normal saline group were markly increased, however, the changes of these two parametere were not so prominent in NGF group. It was suggested that NGF play a role in protecting the spinal cord by maintaining the calcium level of the injured segment.
ObjectiveTo investigate the expression pattern and significance of Sonic Hedgehog (Shh) signaling pathway by observing whether the Shh signaling pathway components express in the adult rat after spinal cord injury (SCI).
MethodsSixty-four healthy male Sprague-Dawley rats were randomly divided into normal group (group A, 8 rats), sham group (group B, 8 rats), and SCI group (group C, 48 rats). In group A, the rats served as controls without any treatment; a decompressive laminectomy was performed on T7-9 levels without SCI in group B; and modified Allen's method was used to make SCI model in group C. Basso Beattie Bresnahan (BBB) scale was used to assess the hind limb motor function at 12 hours, 1 day, 3 days, 7 days, 14 days, and 21 days after SCI; the immunofluorescence staining, real-time PCR, and Western blot were performed to detect the mRNA and protein expression levels of Shh and Glioma-associated oncogene homolog-1 (Gli-1) in SCI zone.
ResultsThe BBB score slowly increased with time in group C, but the scores at each time point in group C were significantly lower than those in group A and group B (P<0.05). The results of immunofluorescence staining showed that Shh and Gli-1 rapidly increased after SCI in astrocytes. Real-time PCR and Western blot showed that the relative expression levels of Shh and Gli-1 mRNA and protein were gradually increased in group C and reached a maximum at 7 days. In addition, the relative expression levels of Shh and Gli-1 mRNA and protein in group C were significantly higher than those in group A and group B (P<0.05). On the other hand, compared with group A, the expression of Gli-1 protein was reduced in the cytoplasm but increased in nucleus in group C.
ConclusionAstrocytes synthesize and secrete Shh and Gli-1 signaling molecules after SCI, both Shh and Gli-1 significantly up-regulate and exhibit dynamic changes, which suggests Shh signaling pathway may be involved in nerve cell regeneration after SCI.
Objective To explore the expression of nestin and glialfibrillary acidic protein (GFAP) at different time and sites after spinal cord injury in adult rats. Methods Seventy-two adult Sprague-Dawley rats, aging 8 weeks and weighing from 180 to 220 g, were randomly divided into 11 experimental groups(66, n=6) and 1 control group(n=6). In the experimental groups, the rat spinal cord injury models were established by aneurysm clip compression, and the expression and proliferation of nestin and GFAP at different time（1 day,3 days, 5 days, 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks,7 weeks and 8 weeks）and at different sites(injured site and adjacent site) were observed with toludine blue staining, immunofluorescent staining and the analytical system of photographs. In control group, the same site of the rat spinal cord was exposed without aneurysm clip compression. The same preparation and examination were done as the experimental groups. Results Toluidine blue staining results showed thatcontour of neurite and pericaryon were distinct and nucleus were deep blue in normal control rats. One day after injury, the number of big and medium-sized neuron decreased obviously; neurite was deep blue with clouding Nissl bodies and ellipse or triangular typed nucleus. In the normal control group, the expression of nestin was hardly seen except ependymal cells of central canal, and the low expression of GFAP was seen. In the experimental groups, the nestin and GFAP expressions increased obviously in the injured sites and adjacent sites 24 hours after injury, reached the peak value after 3-7 days and followed by gradual decrease. There were statistically significant differences in the nestin and GFAP expressions between theexperimental groups and the control group.Conclusion The aboveresults suggestthat spinal cord injury can induce the expression of nestin and GFAP. There is a positive correlation between nestin expression and the proliferation of the reactive astrocytes.
In order to study the prophylactic and therapeutic effect of methyl-prednisolone (MP) on traction injury of spinal cord, 48 rabbits were divided into four groups randomly. According to decreasing amount of the amplitude of P1-wave, 50% reduction lasted for 5 min and 10 min with MP as experimental group, and 50% 5 min and 10 min with NS as control, the changes of amplitudes were monitored by, and the function of the spinal cord was assessed. The amounts of MDA and SOD of the spinal cord tissue were determined and the pathomorphological changes of the spinal cord were observed. The results showed that in the experimental groups, the recovery of P1-wave was quicker, the Tarlov and Molt value were decreased, the density of gray matter of the anterior horn and the myelinated nerve fiber of white matter of 100 microns diameter were higher, the SOD and MDA was decreased and the degenerative and necrotic degree of neuron and nerve fiber were milder. Where in the control groups all the above items were just on the opposite. The conclusions list as follows: the application of MP before operation of spinal deformity might prevent traction injury of the spinal cord during operative correction of spinal deformity, and could also minimized the secondary damage to spinal cord from traction injury if MP was used in time. The action to MP were summarized as improving the microcirculation, inhibiting the hyperoxidation of lipid and accelerating the recovery of SCEP.
Objective To transplant intravenously human brain-derived neurotrophic factor (hBDNF) genemodified bone marrow mesenchymal stem cells (BMSCs) marked with enhanced green fluorescent protein (EGFP) to injured spinal cord of adult rats, then to observe the viabil ity of the cells and the expressions of the gene in spinal cord, as well as theinfluence of neurological morphological repairing and functional reconstruction. Methods Ninety-six male SD rats weighing (250 ± 20) g were randomly divided into 4 groups: hBDNF-EGFP-BMSCs transplantation group (group A, n=24), Ad5-EGFPBMSCs transplantation group (group B, n=24), control group (group C, n=24), and sham operation group (group D, n=24). In groups A, B, and C, the spinal cord injury models were prepared according to the modified Allen method at the level of T10 segment, and after 3 days, 1 mL hBDNF-EGFP-BMSCs suspension, 1 mL Ad5-EGFP-BMSCs suspension and 1 mL 0.1 mol/L phosphate buffered sal ine (PBS) were injected into tail vein, respectively; in group D, the spinal cord was exposed without injury and injection. At 24 hours after injury and 1, 3, 5 weeks after intravenous transplantation, the structure and neurological function of rats were evaluated by the Basso-Beattie-Bresnahan (BBB) score, cortical somatosensory evoked potential (CSEP) and transmission electron microscope. The viabil ity and distribution of BMSCs in the spinal cord were observed by fluorescent inverted phase contrast microscope and the level of hBDNF protein expression in the spinal cord was observed and analyzed with Western blot. Meanwhile, the expressions of neurofilament 200 (NF-200) and synaptophysin I was analyzed with immunohi stochemistry. Results After intravenous transplantation, the neurological function was significantly improved in group A. The BBB scores and CSEP in group A were significantly higher than those in groups B and C (P lt; 0.05) at 3 and 5 weeks. The green fluorescence expressions were observed at the site of injured spinal cord in groups A and B at 1, 3, and 5 weeks. The hBDNF proteinexpression was detected after 1, 3, and 5 weeks of intravenous transplantation in group A, while it could not be detected in groups B, C, and D by Western blot. The expressions of NF-200 and synaptophysin I were ber and ber with transplanting time in groups A, B, and C. The expressions of NF-200 and synaptophysin I were best at 5 weeks, and the expressions in group A were ber than those in groups B and C (P lt; 0.05). And the expressions of NF-200 in groups A, B, and C were significantly ber than those in group D (P lt; 0.05), whereas the expressions of synaptophysin I in groups A, B, and C were significantly weaker than those in group D (P lt; 0.05). Ultramicrostructure of spinal cords in group A was almost normal. Conclusion Transplanted hBDNF-EGFP-BMSCs can survive and assemble at the injured area of spinal cord, and express hBDNF. Intravenous implantation of hBDNF-EGFP-BMSCs could promote the restoration of injured spinal cord and improve neurological functions.
Objective To investigate the effect of chondroitinase ABC (ChABC) on the expression of growth associated protein 43 (GAP-43) and gl ial fibrillary acidic protein (GFAP) after spinal cord injury (SCI) in rats. Methods A total of 150 adult female SD rats, weighing 250-300 g, were randomly divided into ChABC treatment group (group A), sal ine treatment group (group B), and sham operation group (group C) with 50 rats in each group. In groups A and B, the rats were made the SCI models and were treated by subarachnoid injection of ChABC and sal ine; in group C, the rats were not treated as a control. At 1, 3, 7, 14, and 21 days after operation, the Basso, Beattie, and Bresnahan (BBB) score system was used toevaluate the motion function, and immunofluorescent histochemical staining was used to observe the expressions of GAP-43 and GFAP. Results At different time points, the BBB scores of groups A and B were significantly lower than those of group C (P lt; 0.05); there was no significant difference in BBB score between groups A and B after 1, 3, and 7 days of operation (P gt; 0.05), but the BBB score of group A was significantly higher than that of group B after 14 and 21 days of operation (P lt; 0.01). At different time points, the GAP-43 and GFAP positive neurons of groups A and B were significantly higher than those of group C (P lt; 0.05). After 14 and 21 days of operation, the GAP-43 positive neurons of group A were more than those of group B (P lt; 0.01). After 7, 14, and 21 days of operation, the GFAP positive neurons of group A were significantly less than those of group B (P lt; 0.01). Conclusion ChABC can degrade gl ial scar, improve the microenvironment of the injured region and enhance the expression of GAP-43, which promotes axonal growth and extension.
Objective
To discuss the clinical value of whole spine magnetic resonance imaging (WSMRI) in practice of neurosurgical spinal surgery.
Method
A total of 70 cases examined using WSMRI between January 2015 and December 2016 were collected and analyzed retrospectively.
Results
All patients got clear images of WSMRI. Eighteen cases got important information, including spinal variation (1 case), multiple lesions (3 cases), combined lesions (6 cases) and large range multi-segmental lesions (8 cases), which were missed by single-segment MRI .
Conclusions
WSMRI can show all the spine, spinal cord and surrounding tissue in one image at one time. It has high clinical value because of its accurate positioning, comprehensiveness, time saving, and low rate of misdiagnosis and missed diagnosis.
The capacity of embryonic spinal cord tissue in the repair of injured structure of spinal cord has been noted for years. In order to investigate the embryonic spinal cord graft in the repair of motor function of injured spinal cord, the embryonic spinal cord tissue was transplanted to the hemisection cavity in spinal cord in adult rat. One hundred adult Wistar Rats were used to simulate the hemisectional injury of spinal cord by drilling 2-3 mm cavity in lumbar enlargement. Sixty rats were treated with rat embryonic spinal cord tissue grafting while the other forty were chosen as control. The outcome was evaluated according the combined behavioural score (CBS) and motor evoked potential (MEP) in the 1, 2, 4 and 12 weeks. The grafting group was superior to the control as assessed by CBS (P lt; 0.05), especially within 4 weeks. (P lt; 0.01). The restoration of the latent peak of early wave(P1, N1) was better in the grafting group, too. This suggested that embryonic spinal cord graft could improve the recovery of motor function of injured spinal cord in adult rat. The effect of the embryonic spinal cord tissue graft might be concerned with its secretion of several kinds of neurotrophic factors, nerve growth factor, nerve transmitted factor, or adjustment of hormone.
Objective To investigate the feasibil ity of establ ishment of physiological micturition reflex arc by simultaneously reconstructing the sensory and the motorial nerve of atonic bladder after spinal cord injury. Methods Eight 1-year-old Beegle male canine were selected, weighing 7-12 kg. The left side was the experimental side, while the right side wasthe control side. Epidural microanastomosis of vertebral canal of the left L7 ventral root to S2 ventral root and L7 dorsal root to S2 dorsal root was performed to reconstruct the sensory and the motorial function of atomic bladder. The right side was used as a control without treatment. The new motor-to-motor, and sensory-to-sensory physiological bladder reflex pathway were establ ished after 12 months of axonal regeneration. Then S1-4 segmental spinal cord was destroyed for preparation of complete paraplegia. The electrophysiological examination and the bladder pressure were detected before and after paraplegia. The canine micturition was observed for 3 months after paraplegia. Nurohistological observation was performed after canine sacrifice. Results Of 8 canine, 7 canine survived. After paraplegia, canines displayed urinary incontinence and frequent micturition at first, nocturnal continence was achieved gradually without frequent micturition after 1 month. Urinary infection at different degrees occurred in 3 canines and was controlled after Norfloxacin was administered orally. The bladder pressure increased to (1.00 ± 0.13) kPa, (0.90 ± 0.12) kPa after trains of stimulation (300 mV, 0.3 ms, 20 Hz, 5 seconds) of S2 dorsal root at the experimental side before and after paraplegia respectively, showing no significant difference (P gt; 0.05). It increased to (1.90 ± 0.10) kPa after the same train of stimulation of S2 dorsal root at control side. There was significant difference between the experimental side and the control side (P lt; 0.01). Single stimulation (300 mV, 0.3 ms) of the S2 dorsal root at the experimental side resulted in evoked potentials recorded from the left S2 ventral root before and after paraplegia. Before and after paraplegia, the ampl itudes of the evoked potentials were (0.68 ± 0.11) mV and (0.60 ± 0.08) mV respectively, showing no significant difference (P gt; 0.05). It was (1.21 ± 0.13) mV while stimulating at the control side. There was significant difference between the experimental side and the control side (P lt; 0.01). Neurofibra of L7 dorsal and ventral root grew into S2 dorsal and ventral root on tissue sl ice under l ight microscope. Conclusion Reconstruction of the bladder physiological micturition reflex arc is feasible by anastomosis of sacral dorsal and ventral root below injured spinal plane with the suprasacral survival dorsal and ventral root above the plane respectively for restoration of atonic bladder after spinal cord injury.
