Objective To review researches on bone defect repaired by different kinds of bio-derived bone. Methods The recent relevant literatures were extensively investigated. Preparation of bio-derived bone and effect of bone defect repair were reviewed.Results Theallogeneic and xenogeneic bone treated by different physicochemical method werenot only the substitution for bone but also the scaffold material co-cultured with seed cells to reconstruct tissue engineered bone. Conclusion The tissue engineered bioderived bone is a breakthrough for treatment of bone defect.
Objective To investigate the effectiveness of radical debridement, reconstruction with bone allograft, and pedicle screw-rod internal fixation via combined anterior and posterior approach in the treatment of lumbosacral tuberculosis. Methods Between January 2005 and May 2010, 16 patients with lumbosacral tuberculosis were treated. Radical debridement wasperformed via extraperitoneal approach, then tricortical il iac bone allograft was placed and pedicle screw-rod internal fixation was used to reconstruct the spinal column. There were 12 males and 4 females aged 38-65 years (mean, 48 years). The disease duration ranged from 6 to 24 months (mean, 10 months). The main cl inical symptom was persistent pain in lumbosacral area. The involved segments included L4, 5 (3 cases), L5, S1 (8 cases), and L4-S1 (5 cases). The lumbosacral angle was 18-32° (mean, 22°). The erythrocyte sedimentation rate (ESR) was 15-55 mm/1 hour (mean, 25 mm/1 hour). All the patients were given antituberculosis chemotherapy for 12 months after operation. Results The operation time was 120-240 minutes (mean, 180 minutes). The amount of bleeding was 300-600 mL (mean, 420 mL). All wounds healed by first intention, and no relative compl ication occurred. All 16 cases were followed up 12-24 months (mean, 16 months). No recurrence occurred and ESR recovered to normal. Persistent pain in lumbosacral area and radicular pain in lower extremities disappeared. The X-ray films demonstrated that bony fusion was obtained in all patients at 8-12 months postoperatively. The lumbosacral angle was 16-31° (mean, 21°) at last follow-up. Conclusion The extraperitoneal approach can provide direct and safe access to the lesion. The structural il iac bone allograft and posterior instrumentation could reconstruct effectively the stabil ity of the lumbosacral junction.
Objective To investigate tissue engineered spinal cord which was constructed of bone marrow mesenchymal stem cells (BMSCs) seeded on the chitosan-alginate scaffolds bridging the both stumps of hemi-transection spinal cord injury (SCI) in rats to repair the acute SCI. Methods BMSCs were separated and cultured from adult male SD rat. Chitosan-alginate scaffold was produced via freeze drying, of which the structure was observed by scanning electron microscope (SEM) and the toxicity was determined through leaching l iquor test. Tissue engineered spinal cord was constructed by seeding second passage BMSCs on the chitosan-alginate scaffolds (1 × 106/mL) in vitro and its biocompatibil ity was observed under SEM at 1, 3, and 5 days. Moreover, 40 adult female SD rats were made SCI models by hemi-transecting at T9 level, and were randomly divided into 4 groups (each group, n=10). Tissue engineered spinal cord or chitosan-alginate scaffolds or BMSCs were implanted in groups A, B, and C, respectively. Group D was blank control whose spinal dura mater was sutured directly. After 1, 2, 4, and 6 weeks of surgery, the functional recovery of the hindl imbs was evaluated by the Basso-Beattie-Bresnahan (BBB) locomotor rating score. Other indexes were tested by wheat germ agglutinin-horseradish peroxidase (WGA-HRP) retrograde tracing, HE staining and immunofluorescence staining after 6 weeks of surgery. Results Chitosan-alginate scaffold showed three-dimensional porous sponge structure under SEM. The cells adhered to and grew on the surface of scaffold, arranging in a directional manner after 3 days of co-culture. The cytotoxicity of chitosan-alginate scaffold was in grade 0-1. At 2, 4, and 6 weeks after operation, the BBB score was higher in group A than in other groups and was lower in group D than in other groups; showing significant differences (P lt; 0.05). At 4 and 6 weeks, the BBB score was higher in group B than in group C (P lt; 0.05). After 6 weeks of operation, WGA-HRP retrograde tracing indicated that there was no regenerated nerve fiber through the both stumps of SCI in each group. HE and immunofluorescence staining revealed that host spinal cord and tissue engineering spinal cord l inked much compactly, no scar tissue grew, and a large number of neurofilament 200 (NF-200) positive fibers and neuron specitic enolase (NSE) positive cells were detected in the lesioned area in group A. In group B, a small quantity of scar tissue intruded into non-degradative chitosan-alginate scaffold at the lesion area edge, and a few of NSE flourescence or NF-200 flourescence was observed at the junctional zone. The both stumps of SCI in group C or group D were filled with a large number of scar tissue, and NSE positive cells or NF-200 positive cells were not detected. Otherwise, there were obviously porosis at the SCI of group D. Conclusion The tissue engineered spinal cord constructed by multi-channel chitosan-alginate bioscaffolds and BMSCs would repair the acute SCI of rat. It would be widely appl ied as the matrix material in the future.
Objective To investigate the influence of different transplantating times on the survival and immigration of the bone marrow mesenchymal stem cells (BMSCs) in injured spinal cord by subarachnoid administration, and to evaluate the most optimal subarachnoid administration times for BMSCs. Methods Eight adult male rats (weighing 120 g) were used to isolate BMSCs that were cultured, purified and labeled with Hoechst 33342 in vitro. Another 75 adult Wistar rats (weighing 220 g) were made the spinal cord injury (SCI) models at T9,10 level according to the improved Allen’s method and were randomly divided into 5 groups (groups A, B, C, D, and E, n=15). The labeled BMSCs at 1 × 107/mL 0.1 mL were injected into subarachnoid space of the rats via a catheters under the subarachnoid space in groups A (one time at 1 week), B ( two times at 1 and 3 weeks), C (3 times at 1, 3, and 5 weeks) and D (5 times at 1, 3, 5, 7, and 9 weeks) and 0.2 mL phosphate-buffered sal ine (PBS) was injected in group E (5 times at 1, 3, 5, 7, and 9 weeks) as blank control. The neurological functions were evaluated using the Basso-Beattie-Bresnahan (BBB) scale 1, 3, 5, 7, 9, and 12 weeks after transplantation. The migration, survival, differentiation, and histomorphological changes of BMSCs were observed by HE, immunohistochemistry, and fluorescence microscopy. Results At 3 weeks after injury, there were significant differences in the BBB scores between group E and groups A, B, C, D (P lt; 0.01), and between groups A, B and groups C, D (P lt; 0.01). At 7, 9, and 12 weeks, the BBB scores were significantly higher in groups C and D than in groups A and B (P lt; 0.01), and in group B than in group A (P lt; 0.01). There were no significant differences in the BBB scores between groups C and D (P gt; 0.05). The fluorescence microscopy showed that the transplanted BMSCs survived and grew in the injured region at 3 weeks after injury and as time went on, the transplanted cells gradually decreased in group A; in groups B, C, and D, BMSCs count reached the peak values at 5 and 7 weeks and then gradually decreased. At 12 weeks, the survival BMSCs were significantly more in groups C and D than in groups A and B (P lt; 0.01). HE staining showed that the formation of cavity was observed in each group at 3 weeks after injury and the area of cavity gradually decreased in groups A, B, C, and D. At 12 weeks, the area of cavity was the miximal in groups C and D, moderate in groups A and B, and the maximal in group E. The immunohistochemistry staining indicated that the expression of NF-200 was more intense in groups C and D than in groups A and B. The expression of NF-200-positive fibers was more intense in group C. Conclusion Multiple administration of BMSCs promotes the restoration of injured spinal cord and improves neurological functions, and three times for BMSCs transplantation is best
Objective To investigate the effectiveness on the re-fracture of the femur with occult infection by using non-contact locking plate which was placed under the deep fascia. Methods Clinical data of 12 cases of occult infective re-fracture after femoral fracture operation were retrospectively analysed between January 2010 and December 2014. There were 8 males and 4 females with an age of 28-69 years (mean, 42.8 years). Femur re-fractured in 5 cases after 3 days to 4 weeks (mean, 10.6 days) of removal of internal fixation, including 4 cases of plate fixation and 1 case of intramedullary nail fixation; femur in 7 cases re-fractured because of breakage of internal fixator after 7-16 months (mean, 9.3 months) of internal fixation, including 5 cases of plate fixation and 2 cases of intramedullary nail fixation. The tissues near the fracture were collected for bacteria culturing and pathological examining. All the patients were treated by debriding the site of the fracture, bridging with the non-contact locking plate, and transplanting with granulated cancellous bone autograft. Intravenous infusion of antibiotics were used for 2-3 weeks after operation and oral administration for 4 weeks. The X-ray films were taken regularly and the function of the knees were evaluated by the Hospital for Special Surgery (HSS) score system. Results The results of bacteria culturing were positive in 8 patients and negative in 4 patients, and the pathological results of all the patients were confirmed to be chronic bone infection. All the fractures healed with no signs of exudation and ulceration of the incisions. The 12 patients were followed up 18-36 months (mean, 29.6 months). The fracture healed well and no re-fracture occurred. The fracture healing time was 14-22 weeks (mean, 18 weeks). At last follow-up, the function of the knee joint was excellent in 9 cases and good in 3 cases according to HSS score system. Conclusion The treatment of re-fractures after femur fracture operation needs to determine whether there is an occult infection, and non-contact locking plate placed under the deep fascia is an effective way for the re-fracture.
ObjectiveTo investigate the effectiveness on the distal radius deformity and bone defect after trauma by using Ilizarov external fixator.MethodsThe clinical data of 9 patients of post-traumatic distal radius deformity with bone defect treated by Ilizarov technique between January 2012 and December 2016 were retrospectively analyzed. There were 7 males and 2 females with an average age of 25.6 years (range, 11-46 years). Of the 9 cases, 4 were radial baseball hand deformity with large bone defect, 4 were short deformity of distal radius, 1 was distal radius deformity with radial deflection and pronation deformity, all with distal dislocation of the distant radial-ulnar joint. The time from injury to operation was 6 months to 6.2 years (mean, 1.5 years). The bone defect was 1.4-6.8 cm (mean, 3.6 cm). After complete debridement, the forearm was fixed with Ilizarov external fixator. At 7 days after operation, bone transport or bone lengthening was performed at the rate of 0.8-1 mm/d, 4 times a day, the deformity was slowly corrected and the bone defect was repaired. According to the loss of palmar tilt angle and ulnar tilt angle measured before operation, the position of distal radial articular surface was gradually adjusted in the course of moving or prolonging, so as to restore palmar tilt angle and ulnar tilt angle as far as possible.ResultsAll wounds healed by first intention and no leakage or rupture occurred. All the 9 patients were followed up 15-36 months (mean, 23 months). All the radius defects healed and the distal deformity was corrected, the healing time was 92.4-138.6 days (mean, 104.7 days); the external fixation index was 32.6-51.1 days/cm (mean, 40.2 days/cm). After 2 months of external fixator removal, the wrist joint flexion was (42.6±3.1)°, the wrist dorsum extension was (48.5±4.7)°, the palm inclination angle was (11.5±1.3)°, and the ulnar deviation angle was (21.2±3.7)°; the elbow flexion was (128.2±6.4)°, the elbow extension was (3.2±2.1)°, the forearm pronation was (71.5±4.3)°, and the forearm rotation was (38.2±6.5)°; the wrist and elbow joint extension and forearm rotation were significantly improved when compared with preoperative values (P<0.05). At last follow-up, wrist function was assessed according to Gartland-Werley standard, the results were excellent in 3 cases, good in 5 cases, and fair in 1 case. Four cases had pinhole infection, and were cured after anti inflammatory dressing change or replacement of needles; 3 cases did not heal at the bone junction, and were healed after bone grafting; 4 cases deviated from the radial force line, and the deformity was corrected after adjusting the needle.ConclusionIlizarov technique can correct deformity and reconstruct bone defect of the post-traumatic distal radius simultaneously, so it is a good method to treat this kind of disease.
ObjectiveTo clarify the structure and biomechanical characteristics of the dura mater of the cervical, thoracic, and lumbar segments of sheep, in order to provide a theoretical reference for the study of artificial dura mater.MethodsFive adult male white sheep were sacrificed. The dura mater of C5, T10, and L3 planes were obtained. The histological HE staining was used to observe the internal structure and the thickness of dura mater; the inner and outer surfaces morphology of the dura was observed by scanning electron microscopy (SEM); transmission electron microscopy (TEM) was used to observe the internal structure of dura mater and to measure the diameter of collagen fibers in each part of dura mater. The dura mater of C6, C7, T11, T12, L4, and L5 planes were taken for uniaxial biomechanical test, and modulus of elasticity, tensile strength, and elongation at break were measured.ResultsHE staining showed that the thickness of the cervical, thoracic, and lumbar dura mater gradually decreased, and the thickness of the dura mater was (268.19±15.91), (198.16±27.25), (103.74±21.54) μm, respectively, and the differences were significant (P<0.05). SEM observation showed that there were more collagen fibers and fewer cells on the inner surface of the dura mater, while more cells were distributed on the outer surface, and the cells on the inner and outer surface were stretched along the longitudinal axis. TEM observation showed that the collagen fibers in the dura mater were interlaced and arranged in layers. The collagen fibers in the lamina were arranged in the same direction, and the collagen fibers between the lamina were arranged vertically. The diameters of collagen fibers in the cervical, thoracic, and lumbar dura mater were (68.04±21.00), (64.54±20.64), (60.36±19.65) nm, respectively, and the differences were not significant (P>0.05). Uniaxial biomechanical tests results showed that there was no significant difference in modulus of elasticity, tensile strength, and elongation at break between the axial and transverse dura mater of the cervical dura mater (P>0.05); the axial data of thoracic and lumbar segments were significantly larger than the transverse data (P<0.05). The axial modulus of elasticity, tensile strength, and elongation at break of the dura mater of the cervical, thoracic, and lumbar dura mater were significantly different (P<0.05) from the transverse ones, and showing a decreasing trend. Among them, the ratio of axial and transverse modulus of elasticity of cervical and thoracic dura were significantly smaller than that of lumbar segment (P<0.05), and there was no significant difference between cervical segments and thoracic segments (P>0.05).ConclusionThe thickness of dura mater in sheep decreased gradually from head to tail. There are more collagen fibers and fewer cells on the inner surface of dura mater, while the outer surface of dura mater is covered by cells. The collagen fiberboard layers in the dura mater are arranged alternately, and have obvious anisotropic biomechanical characteristics, and the anisotropic biomechanical characteristics get more significant from the head to the tail.
Objective To study the effect of dimethyloxalylglycine (DMOG) on angiogenesis in Choke Ⅱ zone of rats cross-zone perforator flaps and its mechanism. Methods One hundred and twenty-six adult male Sprague Dawley rats were randomly divided into DMOG group, YC-1 group, and control group, with 42 rats in each group. Cross-zone perforator flap model with size of 12 cm×3 cm was made on the back of rats in the three groups. DMOG group was intraperitoneally injected with DMOG (40 mg/kg) at 1 day before operation, 2 hours before operation, and 1, 2, and 3 days after operation; YC-1 group and control group were intraperitoneally injected with YC-1 (10 mg/kg) and the same amount of normal saline at the same time points, respectively. The survival of flap was observed after operation. At 7 days after operation, the survival area of flap in each group was measured and the survival rate of flap was calculated. Flap transmittance test, gelatin-lead oxide angiography, and HE staining were used to observed the angiogenesis in the Choke Ⅱ zone of flaps in each group. Immunohistochemical staining and Western blot were used to detect the expressions of vascular endothelial growth factor (VEGF) and hypoxia-inducible factor 1α (HIF-1α) in Choke Ⅱ zone of flaps in each group. The expressions of VEGF and HIF-1α were also determined by ELISA at 3, 5, and 7 days. Results At 7 days after operation, there was no obvious necrosis at the distal end of the flap in DMOG group, while necrosis occurred in both the control group and YC-1 group, mainly located at the distal end. The flap survival rate of DMOG group was 90.28%±1.37%, which was significantly higher than that of YC-1 group (84.28%±1.45%) and control group (85.83%±1.60%) (P<0.05). DMOG group had more angiogenesis in Choke Ⅱ zone and the vascular structure was clear and complete. In YC-1 group and control group, the vessels in Choke Ⅱ zone was less and the vascular structure was disordered. The number of vessels was (25.56±1.29)/field in the DMOG group, which was significantly higher than that in the YC-1 group [(7.38±0.54)/field] and the control group [(14.48±0.91)/field] (P<0.05). At 3, 5, and 7 days after operation, HIF-1α and VEGF expressions in ChokeⅡzone of DMOG group were significantly higher than those in YC-1 group and control group (P<0.05). ConclusionDMOG can promote angiogenesis in Choke Ⅱ zone, accelerate the early angiogenesis of the flap, improve the microcirculation and blood supply in the potential zone of the flap, reduce the injury of flap ischemia and hypoxia, and increase the survival rate of the flap.