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 study the effectiveness of long segment fixation combined with vertebroplasty (LSF-VP) for severe osteoporotic thoracolumbar compressive fractures with kyphosis deformity. Methods Between March 2006 and May 2012, a retrospective analysis was made on the clinical data of 48 cases of severe osteoporotic thoracolumbar compressive fractures with more than 50% collapse of the anterior vertebral body or more than 40 ° of sagittal angulation, which were treated by LSF-VP in 27 cases (LSF-VP group) or percutaneous kyphoplasty (PKP) in 21 cases (PKP group). All patients suffered from single thoracolumbar vertebral compressive fracture at T11 to L2. There was no significant difference in gender, age, spinal segment, and T values of bone mineral density between 2 groups (P gt; 0.05). The effectiveness of the treatment was appraised by visual analogue scale (VAS), Cobb angle of thoracolumbar kyphosis, height of anterior/posterior vertebral body, and compressive ratio of vertebrae before and after operations. Results The LSF-VP group had longer operation time, hospitalization days, and more bone cement injection volume than the PKP group, showing significant differences (P lt; 0.05). Intraoperative blood loss in LSF-VP group ranged from 220 to 1 050 mL (mean, 517 mL). No pulmonaryor cerebral embolism or cerebrospinal fluid leakage was found in both groups. Asymptomatic bone cement leakage was found in 3 cases of LSF-VP group and 2 cases of PKP group. The patients were followed up for 16-78 months (mean, 41.1 months) in LSF-VP group, and 12-71 months (mean, 42.1 months) in PKP group. No fixation failure such as loosened or broken pedicle screw was found in LSF-VP group during the follow-up, and no re-fracture or adjacent vertebral body fracture was found. Two cases in PKP group at 39 and 56 months after operation respectively were found to have poor maintenance of vertebral height and loss of rectification (Cobb angle was more than 40º) with recurrence of pain, which were treated by second surgery of LSF-VP; another case had compressive fracture of the adjacent segment and thoracolumbar kyphosis at 16 months after operation, which was treated by second surgery of LSF-VP. There were significant differences in the other indexes between each pair of the three time points (P lt; 0.05), except the Cobb angle of thoracolumbar kyphosis, and the height of posterior vertebral body between discharge and last follow-up in LSF-VP group, and except the Cobb angle of thoracolumbar kyphosis and compressive ratio of bertebrae between discharge and last follow-up in PKP group (P gt; 0.05). After operation, the other indexes of LSF-VP group were significantly better than those of PKP group at each time point (P lt; 0.05), except the VAS score and the height of posterior vertebral body at discharge (P gt; 0.05). Conclusion The effectiveness of LSF-VP is satisfactory in treating severe osteoporotic thoracolumbar compressive fractures with kyphosis deformity. LSF-VP can acquire better rectification of kyphosis and recovery of vertebral body height than PKP.
Objective To study the osteogenic effects of a new type of peptides anchored aminated-poly-D, L-lactide acid (PA/PDLLA) scaffold in repairing femoral defect in rats. Methods The PDLLA scaffolds were treated by ammonia plasma and subsequent anchor of Gly-Arg-Gly-Asp-Ser (GRGDS) peptides via amide linkage formation. Thus PA/PDLLA scaffolds were prepared. The bone marrow was harvested from the femur and tibia of 4 4-week-old Sprague Dawley (SD) rats, and bone marrow mesenchymal stem cells (BMSCs) were isolated and cultured by whole bone marrow adherence method. BMSCs-scaffold composites were prepared by seeding osteogenic-induced BMSCs at passages 3-6 on the PA/PDLLA and PDLLA scaffolds. The right femoral defects of 8 mm in length were prepared in 45 adult male SD rats (weighing, 350-500 g) and the rats were divided into 3 groups (n=15) randomly. BMSCs-PA/PDLLA (PA/PDLLA group) or BMSCs-PDLLA (PDLLA group) composites were used to repair defects respectively, while defects were not treated as blank control (blank control group). General state of the rats after operation was observed. At 4, 8, and 12 weeks after operation, general, radiological, histological, micro-CT observations and real-time fluorescent quantitative PCR were performed. Results Two rats died after operation, which was added; the other rats survived to the end of the experiment. At each time point after operation, general and radiological observations showed more quick and obvious restoration in PA/PDLLA group than in PDLLA group; no bone repair was observed in blank control group. The X-ray scores were the highest in PA/PDLLA group, higher in PDLLA group, and the lowest in blank control group; showing significant difference in multiple comparison at the other time (P lt; 0.05) except between blank control group and PDLLA group at 4 weeks (P gt; 0.05). The X-ray scores showed an increasing trend in PDLLA group and PA/PDLLA group with time (P lt; 0.05). Histological and micro-CT observations showed the best osteogenesis in PA/PDLLA group, better in PDLLA group, and worst in blank control group. Comparison between groups had significant differences (P lt; 0.05) in bone mineral density, bone volume/total volume of range of interest, trabecular number, and structure model index. Significant differences (P lt; 0.05) were found in the expression levels of osteogenesis-related genes, such as osteocalcin, alkaline phosphatase, collagen type I, bone morphogenetic protein 2, and osteopontin when compared PA/PDLLA group with the other groups by real-time fluorescent quantitative PCR analysis. Conclusion The PA/PDLLA scaffolds can accelerate the repair of femoral defects in rats.