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BACKGROUND: Human urine-derived stem cells are newly discovered adult stem cells, characterized by rich sources, simple extraction, good proliferative ability and multi-directional differentiation potential. In recent years, human urine-derived stem cells have been used for the repair of neurological functions in urinary diseases, such as stress urinary incontinence and vesicoureteral reflux. OBJECTIVE: To explore the biological characteristics of human urine-derived stem cells and to study their repairing effect in a rat model of spinal cord injury. METHODS: Cell phenotypes of human urine-derived stem cells were detected using flow cytometry, and the immunohistochemical staining was used to identify neuron-like cells differentiated from human urine-derived mesenchymal stem cells. Then, an animal model of spinal cord injury at T9 segment was made by Allen method, and after modeling 24 Sprague-Dawley rats were assigned into spinal cord injury group or cell treatment group (n=12/group). In the cell treatment group, the model rats were injected 2 μL of 1.0×1011/L human urine-derived stem cells, while in the spinal cord injury group, the rats were administered the same volume of L-DMEM containing 10% fetal bovine serum. Basso, Beattie and Bresnahan scores were valued at 1, 10, 20, and 30 days after modeling. Spinal cord samples from all the rats were taken out at 30 days after modeling, and Luxol Fast Blue staining, microglia/macrophages staining and glial fibrillary acidic protein staining were used to value the injured area of the spinal cord and the fluorescence intensity of glial fibrillary acidic protein. RESULTS AND CONCLUSION: (1) Flow cytometry showed high expression on CD29 and CD90, and low expression on CD45 in human urine-derived mesenchymal stem cells. Moreover, human urine-derived mesenchymal stem cells could be induced to differentiating into neuron-like cells in vitro. (2) Basso, Beattie and Bresnahan scores showed no significant difference between the two groups at 1 and 10 days after modeling (P > 0.05), while, at 20 and 30 days after modeling, the scores in the cell treatment group were significantly higher than those in the spinal cord injury group (P < 0.05). (3) Luxol Fast Blue staining showed that the injured area of the spinal cord in the cell treatment group was markedly less than that in the spinal cord injury group (P < 0.05), and the glial fibrillary acidic protein showed lower fluorescence intensity in the cell treatment group than the spinal cord injury group (P < 0.05). To conclude, human urine-derived stem cells can differentiate into neuron-like cells and have therapeutic effects in the rat model of spinal cord injury.
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Objective To compare the therapeutic effects of orthotopic injection and tail vein injection of human amniotic mesenchymal stem cells(hAMSCs)on histological restoration and neurological functions of rats with spinal cord injury. Methods Transected spinal cord injury model in rats was established by transplanting DAPI prelabelled hAMSCs one week after injury.BBB scores were used to evaluate the hindlimb movement of rats. The histological patterns.and morphology of medullary sheath of spinal cord were observed. Results BBB scores in the orthotopic injection group and tail vein injection group were increased gradually from one to six week after hAM-SCs transplantation and reached 6.5 ± 0.5 and 7.12 ± 1.61 respectively 6 weeks after cell transplantation,higher than that of the control group(both P < 0.01). However,there was no statistical significance between the two groups.Histological results indicated that the repair of injured tissue in the orthotopic injection group and tail vein injection group were both better than that in the control group,and there were more vesica and loosened layers forming in the injured spinal cord of rats in the PBS control group as compared with the orthotopic and tail vein transplantation group. Conclusion hAMSCs transplantation through tail vein injection could promote histological restoration and neurological regeneration of rats with spinal cord injury,which has the similar therapeutic effects with hAMSCs orthotopic transplantation.
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Objective To observe the effect of epidural injection with different dose of butorphanol on the rats' neurological function.Methods A PE-530 catheter was inserted into the epidural space of all the SpragueDawley rats (male, weighting 180 ～210 g) at L1-2 level.After three days, a total of 32 rats without any motor dysfunction were randomly divided into 4 groups as follows saline(NS) group (group C, n= 8 )and butorphanol injection (B) group( B1∶ n=8;B2∶ n=8;B3∶ n=8).Rats in group C were epidurally injected NS 30 μl each ,and rats in group B1, B2 and B3 were respectively epidurally injected Butorphanol 60 μg/30μl, 120 μg/30 μl,240 μg/30 μl (all diluted with NS) ,and 1 time per day for5 days.The neurological function of rats was recorded before injection (T0) and 6h after injection on day 1 ～4(T1 ～T 4) and 6h,24h and 72h after injection on day 5 (T5 ～T7) by BBB (BASSO,BEATTIE and BRESNAHAN ) Score and the inclined plane test .Results Compared with group C ,the BBB score and the inclined plane test of group B1 showed no significant difference throughout the experimental period(P＞ 0.05 ).There was also no significant difference at T0 ～ T3 of group B2 and group B3 compared with group C (P ＞ 0.05 ), while at T4, the BBB score ( ( 18.50 ± 2.00 ) points, ( 16.38 ± 2.33 ) points) and the inclined plane test( (58.75 ± 5.17 )°, (59.38 ± 3.20) ° ) of the two groups were both obviously decreased when compared with group C( (21.00 ±0.00) points, (65.00 ±3.78)°, P＜0.05) ,and the same significant differences appeared at T5,T6 and T7 (P ＜ 0.05 ).Conclusion Repeated epidural injection of butorphanol 60 μg have no effect on neurological function of rats,while repeated epidural injection of butorphanol 120 μg and 240 μg could impaire the neurological function.
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Objective:To observe neuroprotective effects of insulin on traumatic spinal cord injury in rat models and discuss its potential mechanisms. Methods:60 SD rats were randomly divided into A group(control group) and B group(insulin group), with 30 rats in each group,The animal SCI model was established according to method Allen's,A group and B group were respectively infused with isotonic Na chloride or 1Iu/kg insulin through abdominal cavity,three times a day,for 7 days. apoptosis neurons were observed on 8 hours,3 days,7 days,14 days,and 28 days after SCI,and tigroid body,neurons,GAP-43 positive cells,and BBB score were measured on 1 day,3 days,7 days,14 days,and 28 days after SCI. Results:Neuron number,GAP-43 positive cells,and BBB score in multitude time points were significantly higher in B group than in A group(P