Objective To explore the influence factors for the functional improvement after the fetal olfactory ensheathing cell (OEC) transplantation for chronic spinal cord injury(SCI). Methods The olfactory bulbs were harvested and trypsinized down to single fetal OEC. They were cultured for 12-17 days prepared for use. From November 2001 to December 2003, a total of 300 patients volunteered for the fetal OEC transplantation, among whom 222 suffered from complete chronicSCI and 78 suffered from incomplete chronic SCI. The procedures were performed on the patients with a disease course ranging from 6 months to 31 years (average 3.1 years) after their injuries. The fetal OEC was transplanted by the form of injections into the spinal cord at the upper and lower ends of the injury site. All the patients were assessed by the ASIA standard before the transplantation and 2-8 weeks after the transplantation. The influence factors including age, sex, duration after the injury, and injury degreesand levels were compared with those in the functional improvement after fetal OEC transplantation. Results The partially-improved neurological functions assessed by the ASIA standard were indicated by the motor scores increasing from 39.1±20.6 to 45.9±20.3 (Plt;0.001), the light touch scores from 51.7±24.9 to 63.4±23.0 (Plt;0.001), and the pin prick scores from 53.0±24.2 to 65.3±22.7(Plt;0.001). There was no significant difference in the functional improvement of the motor, light touch, and pin brick when compared with the age, sex, duration after theinjury, and the injury degrees and levels. The motor scores and light touch scores at the cervical level were higher than the scores at the thoracic level. Conclusion The fetal OEC transplantation can partially improve the neurological functions quickly in treatment of the chronic spinal cord injury. All the influence factors except the motor scores and light touch scores, which were higher at the cervical level than at thoracic level, have no impact on the functional improvement after the fetal OEC transplantation.
OBJECTIVE: To investigate the etiology, pathological mechanism and treatment of cervical fracture-dislocation without spinal cord injury. METHODS: Nine patients with cervical fracture-dislocation without spinal cord injury were male and aged 22 to 63 years. Based on the clinical symptoms and roentgenographic changes, the injury mechanism was analyzed; and the pathological characteristics and treatment principle were put forward. RESULTS: Anterior reduction was employed in all 9 cases. Eight cases were reduced completely while 1 case was reduced partially. After following up 1 to 3 years, 7 cases recovered completely and the other 2 cases relieved their symptoms obviously. No nervous symptoms aggravated during the following-up period. CONCLUSION: Fracture-dislocation of the cervical spine without spinal cord injury has special pathological mechanism. The surgical intervention is needed for solid fixation and complete decompression without any delayed neurosymptoms.
Objective To review the advances in repair of spinal cord injury by transplantation of marrow mesenchymal stem cells(MSCs). Methods The related articles in recent years were extensively reviewed,the biological characteristic of MSCs,the experimental and clinical studies on repair of spinal cord injury by transplantation of MSCs,the machanisms of immigration and therapy and the problems were discussed and analysed. Results The experimental and clinical studies demonstrated that the great advances was made in repair of spinal cord injury by transplantation of MSCs. After transplantation, MSCs could immigrate to the position of spinal cord injury, and differentiate into nervelike cells and secrete neurotrophic factors.So it could promote repair of injuryed spinal cord and recovery of neurologicalfunction. Conclusion Transplantation of MSCs was one of effective ways in repair of spinal cord injury, but many problems remain to be resolved.
Objective To investigate the division, prol iferation and differentiation abil ities of nestin+/GFAP+cell after spinal cord injury and to identify whether it has the characteristic of neural stem cells (NSCs). Methods Twelvemale SD rats, aged 8 weeks and weighing 200-250 g, were randomized into 2 groups (n=6 per group): model group inwhich the spinal cord injury model was establ ished by aneurysm cl ip compression method, and control group in which no processing was conducted. At 5 days after model ing, T8 spinal cord segment of rats in each group were obtained and the gray and the white substance of spinal cord outside the ependymal region around central tube were isolated to prepare single cellsuspension. Serum-free NSCs culture medium was adopted to culture and serum NSCs culture medium was appl ied to induce differentiation. Immunohistochemistry detection and flow cytometry were appl ied to observe and analyze the type of cells and their capabil ity of division, prol iferation and differentiation. Results At 3-7 days after injury, the model group witnessed a plenty of nestin+/GFAP+ cells in the single cell suspension, while the control group witnessed few. Cell count of the model and the control group was 5.15 ± 0.71 and 1.12 ± 0.38, respectively, indicating there was a significant difference between two groups (P lt; 0.01). Concerning cell cycle, the proportion of S-phase cell and prol iferation index of the model group (15.49% ± 3.04%, 15.88% ± 2.56%) were obviously higher than those of the control group (5.84% ± 0.28%, 6.47% ± 0.61%), indicating there were significant differences between two groups (P lt; 0.01). In the model group, primary cells gradually formed threedimensional cell clone spheres, which were small in size, smooth in margin, protruding in center and positive for nestin immunofluorescence staining, and large amounts of cell clone spheres were harvested after multi ple passages. While in the control group, no obvious cell clone spheres was observed in the primary and passage culture of single cell suspension. At 5 days after induced differentiation of cloned spheres in the model group, immunofluorescence staining showed there were a number of galactocerebroside (GaLC) -nestin+ cells; at 5-7 days, there were abundance of β-tubul in III-nestin+ and GFAP-nestin+ cells; and at 5-14 days, GaLC+ ol igodendrocyte, β-tubul in II+ neuron and GalC+ cell body and protruding were observed. Conclusion Nestin+/GFAP+ cells obtained by isolating the gray and the white substance of spinal cord outside the ependymal region around central tube after compressive spinal cord injury in adult rat has the abil ity of self-renewal and the potential of multi-polarization and may be a renewable source of NSCs in the central nervous system.
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.
Objective To explore the factors to affect severity of hyperextension injury of the cervical spinal cord (HEICSC). Methods Forty-five patients with HEICSC, 35 males and 10 females, aged 27-67 years old (mean 48.2 years old), were retrospectively analyzed. The disease course was 30 minutes to 16 days. According to modified Frankel grading, there were 6 cases of grade A, 8 cases of grade B, 16 cases of grade C and 15 cases of grade D. Spinal cord injuries (SCI) segments were determined according to SCI plane and high signal change (HSC) in spinal cord on MR images. The whole or large part of HSC segments were supposed to be main injured spinal cord segments (MISCSs) and the staccato or patchy HSC ones were supposed to be common injured spinal cord segments (CISCSs). When the external force acting on head or face suffered was larger, the force produced during high-speed movement or forehead and/or face had severe contused and/or) lacerated wound, the force was defined severe traumatic strength, whereas the reverse was true for sl ight traumatic strength. According to signal magnitude of the cervical discs on T2-weighted MR images, degeneration of cervical discs and cervical vertebras were classified into 5 grades: grade 0-4. Cervical spinal stenosis were graded to 5 grades according to the width of anterior or posterior cerebrospinal fluid layer to spinal cord on T2-weighted MR images and compressed degree of spinal cord on T1-weighted MR images. The influence of traumatic strength, cervical spinal degeneration or cervical spinal stenosis on SCI were explored. Results Among the 45 cases, 12 cases were caused by sl ight traumatic strength, 33 cases were caused by severe one. The cervical spinal cord was injuried more sl ightly and the patients were older in the sl ight traumatic strength cases than in the severe ones (P lt; 0.05). The number of MISCSs were 45 in 40 cases and the 25 segments were located at C3, 4 level. The number of CISCSs were 39 in 21 cases. All the cervical vertebraes of the 45 patients had degenerated. The most were in grade 3 in 22 patients and the severest degenerative segments were mostly located in C5,6 discs in 35 ones. The number of the MISCSs in different degenerative grades of discs was 0 in grade 0, 9 in grade 1, 20 in grade 2, 14 in grade 3, and 2 in grade 4. The ratios of the segment number of injuried spinal cord to the segment number of spinal stenosis in every grade of stenosis were 1/62 in grade 0, 2/11 in grade 1, 27/52 in grade 2, 33/33 in grade 3, 21/22 in grade 4. Conclusion Three main factors including the magnitude of traumatic strength, the degree of instabil ity of cervical vertebrae and the degree of cervical stenosis contribute to development and progress of HEICSC.
Objective To establish the artificial bladder reflex arc by the normal body reflex pathway above the horizon of spinal cord injury to reinnervate the flaccid bladder and restore bladder micturition function. Methods An intradural microanastomosis was performed on the L6 ventral root tothe S2 ventral root. After axonal regeneration,the “patellar ligament-spinal cord center-bladder” reflex pathway was reestablished. A longterm function of the reflex arc was observed in the nerve electrophysiological experiment, detrusor electromyography experiment, and urodynamic testing 8 months after anastomosis. Results Trains of the stimuli(200 μV,5 ms) in the left L6 dorsal root and the nerve at the anastomosizedsite resulted in motor evoked potential from the disal to the anastomosized site before and after the spinal cord was destroyed horizontally between S1 and S4 segment levels in 2 Beegle dogs.The figure and amplitude of the evoked potential were similar to those of the control and general stability which showed anoninterventional wave. The urodynamic test revealed a rapid increase of the bladder pressure and a minor increase in the abdominal pressure. This showed that the bladder detrusor mainly resulted in the pressure increase.The bladder pressure increased to 60% of the normal on average compared with the controls when resulted in the left L6 dorsal root and the nerve anastomosized site were stinulated. Conclusion The long-term observation by the nerveelectrophysiological experiment, detrusor electromyography experiment, and urodynamic test indicate that the new artificial reflex arc can be established successfully. The somatic motor axons can regenerate into the parasympathetic endoneurial tubes of the autonomic nerve.
Objective To investigate the diagnostic value of MR imaging in cervical spinal canal stenosis combined with spinal cord injury. Methods From August 1998 to May 2008, 41 patients with cervical spinal canal stenosis and spinal cord injury were treated, including 34 males and 7 females aged 32-71 years (average 53.4 years, 27 patients being older than 60 years). Patients’ MRI data were retrospectively analyzed. Injury was caused by fall ing from height in 8 cases, traffic accidentin 19 cases, crush due to heavy objects in 3 cases and other reasons in 11 cases. The time from injury to operation ranged from 2 hours to 3 years. There were 12 cases of anterior spinal cord injury syndrome, 23 of central spinal cord syndrome and 6 of Brown-Sequard syndrome. JOA score of spinal cord function was 3-11 points (average 6.6 points). Results MR imaging diagnosis before operation showed abnormal signal changes within the spinal cord in 37 cases (41 sites), anterior and posterior longitudinal l igaments and discs (APLLD) injury in 28 cases (30 sites) and signal of edema and hematoma signals in anterior surface of cervical spines (EBC) in 34 cases (36 sites). Diagnosis during operation revealed edemas braises, contusions tears of posterior soft tissue in 18 cases (20 sites), appendix fracture in 6 cases (7 sites), formation of EBC in 20 cases (23 sites), APLLD injury in 34 cases (44 sites), intervertebral instabil ity without the rupture of l igament and intervertebral disc in 7 cases (10 sites). Significant difference was evident between the MRI diagnosis before operation and the intraoperative discoveries (P lt; 0.05). Conclusion The MR imaging diagnosis before operation do not correspond to the intraoperative discoveries, indicating that MRI diagnosis fails to make a relatively comprehensive and accurate diagnosis. So it is advisable to make a diagnosis based on cl inical symptoms.
Objective To explore the construction of a canine model of vascularized allogeneic spinal cord transplantation (vASCT) and preliminarily evaluate its therapeutic efficacy for spinal cord injury (SCI). Methods Sixteen female Beagle dogs aged 8-12 months were randomly selected, with 8 dogs serving as donors for the harvesting of spinal cord tissue with a vascular pedicle [dorsal intercostal artery (DIA) at the T10 level and accompanying vein]. The remaining 8 dogs underwent a 1.5-cm-length spinal cord defect at the T10 level, followed by transplantation of the donor spinal cord tissue for repair. Polyethylene glycol (PEG) was applied to both ends to spinal cord graft; then, using a random number table method, the dogs were divided into an experimental group (n=4) and a control group (n=4). The experimental group received immunosuppressive intervention with oral tacrolimus [0.1 mg/(kg?d)] postoperatively, while the control group received no treatment. The operation time and ischemia-reperfusion time of two groups were recorded. The recovery of hind limb function was estimated by Olby score within 2 months after operation; the motor evoked potentials (MEP) was measured through neuroelectrophysiological examination, and the spinal cord integrity was observed through MRI. ResultsThere was no significant difference in the operation time and ischemia-reperfusion time between the two groups (P>0.05). All dogs survived until the completion of the experiment. Within 2 months after operation, all dogs in the control group failed to regain the movement function of hind limbs, and Olby scores were all 0. In the experimental group, the movement and weight-bearing, as well as walking abilities of the hind limbs gradually recovered, and the Olby scores also showed a gradually increasing trend. There was a significant difference between the two groups from 3 to 8 weeks after operation (P<0.05). Neuroelectrophysiological examination indicated that the electrical signals of the experimental group passed through the transplanted area, and the latency was shortened compared to that at 1 month after operation (P<0.05), showing continuous improvement, but the amplitude did not show significant improvement (P>0.05). The control group was unable to detect any MEP changes after operation. MRI examination showed that the transplanted spinal cord in the experimental group survived and had good continuity with normal spinal cord tissue, while no relevant change was observed in the control group. ConclusionThe vASCT model of dogs was successfully constructed. This surgical procedure can restore the continuity of the spinal cord. The combination of tacrolimus anti-immunity is a key factor for the success of transplantation.
ObjectiveTo investigate the expression changes of the receptor activator of nuclear factor-κB ligand (RANKL) in the peripheral blood of patients with aseptic loosening of the implant after total hip arthroplasty (THA) by comparing with that of patients with femoral neck fracture and to analyze the correlation between RANKL expression and aseptic loosening. MethodsBetween January 2008 and January 2013,the peripheral blood were harvested from 58 patients with aseptic loosening of the implant after THA (trial group) and 63 patients with femoral neck fracture (control group).The 2 groups were well matched,with no significant differences in age and gender (P>0.05).The expressions of the RANKL mRNA and RANKL protein were evaluated by quantitative real-time PCR and Western blot respectively.At the same time,the concentration of RANKL was also measured by ELISA. ResultsThe expression of the RANKL mRNA in the trial group was 18.30±1.09,which was significantly higher than that of control group (1.00±0.05)(t=125.390,P=0.000).The relative RANKL protein expression values in trial group and control group were 0.856±0.254 and 0.404±0.102 respectively,showing significant difference (t=13.032,P=0.000).The results of ELISA showed that the concentration of RANKL in trial group [(3.553 5±0.129 7) ng/mL] was significantly higher than that of control group [(1.912 3±0.126 2) ng/mL] (t=18.124,P=0.000). ConclusionThe high RANKL expression in peripheral blood is probably correlated with aseptic loosening of the implant in patients undergoing THA,which possibly is the prognostic factor of aseptic loosening of the implant.