Objective To explore changes in the height and width of the cervical intervertebral foramina of C6,7 before and after the C5,6 discetomy, the replacement or the anterior intervertebral fusion so as to provide the theoretical basis for the clinical practice. Methods Eleven fresh cervical spinal specimenswere obtained from young adult cadavers. The specimens of C5,6 were divided into the integrity group, the discectomy group, the artificial disc replacement group, and the intervertebral fusion group. The range of variety (ROV) of the C6,7 intervertebral foramen dimensions (height, width) before and after the loading tests (0.75, 1.50 Nm) were measured in the 4 groups. Results The C6,7 intervetebral foramen height and width increased significantly during flexion (Plt;0.01) but decreased significantly during extension (Plt;0.01). There was a significantdifference between the two test conditions in each of the 4 groups (Plt;0.01). However, in the two test conditions there was no significant difference in ROV of the C6,7 intervetebral foramen height and width during flexion and extension betweenthe integrity group, the discectomy, and the artificial disc replacement group（Pgt;0.05）, but a significant difference in the above changes existed in the intervertebral fusion group when compared with the other 3 groups (Plt;0.05). In the same group and under the same conditions, the ROV of the C6,7 intervetebral foramen height and width was significantly different in the two test conditions (Plt;0.01). Conclusion The results have indicated thatartificial disc replacement can meet the requirements of the normal cervical vitodynamics. The adjacent inferior cervical intervetebral foramen increases during flexion but decreases during extension. The intervertebral fusion is probably one of the causes for the cervical degeneration or the accelerated degeneration and for the cervical spondylotic radiculopathy and the brachial plexus compression.
OBJECTIVE: To investigate the expression and distribution of xenoantigen in intervertebral disk of Chinese banna minipig inbred line, and to study the availability of xenograft transplantation of intervertebral disk. METHODS: Samples of intervertebral disk were collected from six Banna pigs of 8 to 11-month-old. The fixation, embedment and slice were performed. α-Gal specific binding lection (BSI-B4) were used as affinity reagents and affinity-immunohistochemistry assays (SABC methods and DAB stain) were conducted to detect the expression and distribution of xenoantigen (α-Gal). RESULTS: alpha-Gal was found in chondrocyte cell and chondrocyte-like cell in intervertebral disk which have the positive yellow-stained particulate aggradation. There was no stain in the matrix, elastic fiber and collagen fiber. CONCLUSIONS: The distribution of xenoantigen is locally in the tissue of intervertebral disk and its expression is weak. This suggests that the intervertebral disk of Banna pig may be alternative donor for xenotransplantation.
Objective To investigate diagnosis and treatment of farlateral lumbar disc herniations. Methods The clinical data from 16 patients with far-lateral lumbar disc herniations from January 1999 to January 2004 were retrospectively analyzed. The CT scanning showed that the shadow density of the CT scanning values in the corresponding intra-foramen, extraforamen and all-foramen was as the same as that of the intervertebral disc. Of the 16 patients, 10 were operated on by the interlaminar approach, 3 were operatedon by the laterolaminar approach, 3 were operated on by the combined interlaminal and laterolaminal approach.Results According to the follow-up for 6 monthsto 5 years, excellent results were obtained in 8 patients, good results in 5, and fair results in 3. The postoperative CT examination showed that the space occupying in the foramen or extraforamen of the corresponding segment vanished and the nerve root compression of the identical segment also vanished. Conclusion The lamellar highresolution CT is a better way to diagnose lumbar disc herniation. The operative approach should be chosen according to the position of the intervertebral disc protrusion, pathologic type, and presence or absence of the lesions in the vertebral canal.
Objective To observe ultrastructural changes of the intervertebraldisk in the corresponding area after internal fixation of spinal column. Methods Twenty-four Japanese big ear rabbits were divided into internal fixation of spinal column group (n=12) and control group (n=12). The internal fixation model was made as follows: The spinous processes and erector spinal muscle were exposed and the T10L3 spinous processes and the relevant two-side articular processes under the periosteumwere isolated. With the help of L-shaped Kirschner wires, the steel wire was threaded through the articular of T11,T12,L1 and L2, and were connected with L-shaped Kirschner wries. After 6 months of operation, the following intervertebral disk tissues were observed with transmission electeon microscope: nucleus pulposus, internal annlus fibrosus and external anulus fibrosus of L1 intervertebraldisk. The T12and L2 intervertebal disk surface structure was observedhorizontally and longitudinally with scanning electron microscope, respectively. Results After internal fixation of spinal column, the structural changes of cells in nucleus pulposus and internal annulus fibrosus occurred earlier than that in the external annulus fibrosus. Proteoglycan and special structure were found in nucleus pulposus and matix of annulus fibrosus. However, the forms of special structure in nucleus pulposus and internal layer of annulus fibrosus were different. In the degeneration matrix of intervertebral disc, the proteoglycan particles and special structure were obviously decreased. Conclusion Abnormal stress environment can result in the degeneration of intervertebral disk. There is a regular distribution of the special structure in nucleus pulposus and matrix of annulus fibrosus, which is related to biology behaviour of proteoglycan particles in the degeneration of intervertebral disk.
OBJECTIVE To investigate the therapeutic effect of percutaneous lumbar discectomy (PLD) combined with sodium hyaluronate (SH) injection in the treatment of lumbar intervertebral disc herniation. METHODS Forty-eight patients suffered from lumbar disc herniation were divided into two groups and treated by PLD combined with SH injection into epidural cavity (treatment group) or single PLD (control group) respectively. All patients were followed up for 24 months. The therapeutic effects in both groups were assessed and compared according to Macnab’s criterion. RESULTS The patients in the treatment group got much more significant improvement than those in the control group, with shorter therapeutic course and more safety. CONCLUSION PLD combined with SH injection into epidural cavity is more effective and safety in the treatment of lumbar disc herniation than of pure PLD.
ObjectiveTo investigate the influence of ISOBAR TTL dynamic internal fixation system on degeneration of adjacent intervertebral disc by MRI measurement of lumbar nucleus pulposus volume in treating lumbar degenerative disease after operation. MethodsBetween March 2010 and October 2011, 34 patients with lumbar intervertebral disc herniation (23 cases of paracentral type and 11 cases of lateral type) underwent operation with ISOBAR TTL dynamic internal fixation system for fixation of single segment, and the clinical data were analyzed retrospectively. There were 20 males and 14 females, aged 39-62 years (mean, 47.5 years). The disease duration was 6-18 months (mean, 14 months). Involved segments included L4, 5 in 21 cases and L5, S1 in 13 cases. The X-ray films and MRI images were taken at 6, 12, 18, 24, 36, and 48 months after surgery. Based on X-ray films, the height of intervertebral space was measured using angle bisectrix method. The nucleus pulposus volume was measured based on the MRI scan. The postoperative change of nucleus pulposus volume and intervertebral disc height were used to evaluate the influence of ISOBAR TTL system on degeneration of adjacent intervertebral disc nucleus pulposus. ResultsThirty patients were followed up 48 months. The height of intervertebral space showed no significant difference between at pre-and post-operation (P>0.05). The nucleus pulposus volume increased after operation, showing no significant difference at 6, 12, and 18 months when compared with preoperative value (P>0.05), but significant difference was found at 24, 36, and 48 months when compared with preoperative value (P < 0.05). The height of nucleus pulposus increased after operation but the width was decreased; the values showed no significant difference at 6, 12, and 18 months when compared with preoperative ones, but showed significant difference at 24, 36, and 48 months when compared with preoperative ones (P < 0.05). The diameter of nucleus pulposus at 18, 24, 36, and 48 months after operation was significantly langer than that at preoperation (P < 0.05). ConclusionISOBAR TTL dynamic internal fixation system can prevent or delay the degeneration of intervertebral discs.
Objective To review the study progress of mesenchymal stem cells induced to differentiate intervertebral disc cells Methods The recent related literature was reviewed. The theorical and experimental studies were summarized. Results MSCs had the potential of multidirectional differentiation.International experimental studies indicated the potential of MSCs induced to differentiate intervertebral disc cells. Conclusion MSCs induced to differentiate intervertebral disc cells has the fine prospect.
OBJECTIVE: To review research progress of the relation between growth factor and repair of intervertebral disc. METHODS: The recent articles on growth factor and repair of intervertebral disc were extensively reviewed. The expression of growth factor in intervertebral disc and the effect of growth factor on disc cells were investigated. RESULTS: Some growth factors play roles in the development and degeneration of intervertebral disc. Exogenous growth factor can increase proliferation of disc cells and production of proteoglycans and collagens. Gene of growth factor can be transferred to intervertebral disc cell by adenovirus. CONCLUSION: Growth factor plays an important role in the regulation of development and degeneration of interertebral disc. The above results show that the feasibility of usage of growth factor in the treatment of disc degeneration and in repair and reconstruction of disc.
Objective To evaluate the cell biological features and the effect of transplantation of transforming growth factor β3 (TGF-β3) gene-modified nucleus pulposus (NP) cells on the degeneration of lumbar intervertebral discs in vitro. Methods NP cells at passage 2 were infected by recombinant adenovirus carrying TGF-β3 (Ad-TGF-β3) gene (Ad-TGF-β3 group), and then the cell biological features were observed by cell vital ity assay, the expression of the TGF-β3 protein was determined by Western blot, the expression of collagen type II in logarithmic growth phase was determined by immunocytochemistry. The cells with adenovirus-transfected (Adv group) and the un-transfected cells (blank group) were used as controls. The model of lumbar disc degeneration was establ ished by needl ing L3, 4, L4, 5, and L5, 6 in 30 New Zealand rabbits (weighing 3.2-3.5 kg, male or female). Then Ad-TGF-β3-transfected rabbit degenerative nucleus pulposus cells (100 μL, 1 × 105/ mL, group A, n=12), no gene-modified nucleus pulposus cells (100 μL, 1 × 105/mL, group B, n=12), and phosphatebuffered sal ine (PBS, 100 μL, group C, n=6) were injected into degenerative lumbar intervertebral discs, respectively. L3, 4, L4, 5, and L5, 6 disc were harvested from the rabbits (4 in groups A and B, 2 in group C) at 6, 10, and 14 weeks respectively to perform histological observation and detect the expression of collagen type II and proteoglycan by RT-PCR. Results The viabil ity of nucleus pulposus cells was obviously improved after transfected by recombinant Ad-TGF-β3 gene. At 3, 7, and 14 days after transfected, TGF-β3 expression gradually increased in nucleus pulposus cells. The positive staining of collagen type II was seen in Ad-TGF-β3 group, and the positive rate was significantly higher than that of Adv group and blank group (P lt; 0.05). The disc degeneration in group A was sl ighter than that in groups B and C. The expressions of collagen type II mRNA and proteoglycan mRNA in group A were significantly higher than those in groups B and C at 6, 10, and 14 weeks (P lt; 0.05). Conclusion TGF-β3 can improve the biological activity of NP cells and promote the biosynthesis of collagen type II and proteoglycan in intervertebral discs, alleviate the degeneration of intervertebral discs after transplantation.
Objective Bone marrow mesenchymal stem cells (BMSCs) transplantation can potentially regenerate the degenerated intervertebral disc, with the underlying regenerating mechanism remaining largely unknown. To investigate the potential of human BMSCs protecting nucleus pulposus cells (NPCs) from oxidative stress-induced apoptosis in a coculturesystem, and to illustrate the possible mechanisms of BMSCs transplantation for intervertebral disc regeneration. Methods BMSCs collected by density gradient centrifugation in Percoll solution were cultured and sub-cultured till passage 3, and the surface molecules of CD34, CD45, and CD13 were identified. NPCs were isolated by collagenase digestion and the chondrocyte l ike phenotype was confirmed by morphologic observation after HE staining, inverted phase contrast microscope, proteoglycan, and collagen type II expression after toluidine blue and immunocytochemistry staining. The 3rd passage BMSCs and the 1st passage NPCs were divided into four groups: group A, NPCs (1 × 106 cells) were cultured alone without apoptosis inducing (negative control); group B, NPCs (1 × 106 cells) were co-cultured with BMSCs (1 × 106 cells) with apoptosis inducing; group C, NPCs (1 × 106 cells) were co-cultured with BMSCs (3 × 105 cells) with apoptosis inducing; group D, NPCs (1 × 106 cells) were cultured alone with apoptosis inducing (positive control). After 3 or 7 days of culture or co-culture, the NPCs in groups B, C, and D were exposed to 0.1 mmol hydrogen peroxide for 20 minutes to induce apoptosis. With DAPI staining cellular nucleus, Annexin-V/propidium iodide staining cellular membrane for flow cytometry analysis, the apoptosis of NPCs in each group was studied both qual itatively and quantitatively. Besides, the changes in Bax/Bcl-2 gene transcription and Caspase-3 protein content, were analyzed with semi-quantitative RT-PCR and Western blot. Results BMSCs were successfully isolated and CD34-, CD45-, and CD13+ were demonstrated; after isolated from degenerated intervertebral discs and sub-cultured, the spindle-shaped 1st passage NPCs maintained chondrocyte phenotype with the constructive expressions of proteoglycan and collagen type II in cytoplasm. DAPI staining showed the nucleus shrinkage of apoptosis NPCs. Co-cultured with BMSCs for 3 days and 7 days, the apoptosis rates of NPCs in groups B (29.26% ± 8.90% and 18.03% ± 2.25%) and C (37.10% ± 3.28% and 13.93% ± 1.25%) were lower than that in group D (54.90% ± 5.97% and 26.97% ± 3.10%), but higher than that of groupA (15.67% ± 1.74% and 8.87% ± 0.15%); all showing significant differences (P lt; 0.05). Besides, semi-quantitative RT-PCR showed Bcl-2 gene transcription up-regulated (P lt; 0.05) and no significant change of Bax (P gt; 0.05); Western blot result showed that the Caspase-3 protein expression of groups B and C was lower than that of group D, and was higher than that of group A; all showing significant differences (P lt; 0.05). Conclusion In a co-culture system without direct cellular interactions, the oxidative stress-induced apoptosis of human NPCs was amel iorated by BMSCs. The enhanced anti-apoptosis abil ity of NPCs preconditioned by co-culturing with BMSCs might come from the decreased Bax/Bcl-2 gene transcription ratio.