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.
ObjectiveTo explore the effectiveness of nano-hydroxyapatite/polyamide-66 (n-HA/PA66) Cage in interbody fusion for degenerative lumbar scoliosis.MethodsA retrospective analysis was designed and conducted for 43 patients, who underwent posterior decompression and n-HA/PA66 Cage interbody fusion with correction of deformity between January 2013 and June 2016. Eighteen cases were single-level fusion (single-level group) and 25 cases were double-level fusion (double-level group). There was no significant difference in gender, age, body mass index, direction of convex, degree of apical rotation, fusion level, the number of osteoporotic patients, pre-operative intervertebral height of fusion segments, coronal Cobb angle, visual analogue score (VAS), and modified Oswestry Disability Index (ODI) between 2 groups (P>0.05). The operation time, intraoperative blood loss, postoperative drainage, hospital stay, and complications of the operation were recorded. Modified ODI, VAS score, and MacNab criteria were adopted to assess clinical outcomes. Radiographic indexes, including intervertebral height of fusion segments, coronal Cobb angle, disc insertion depth, and the bone graft fusion rate, were also evaluated.ResultsThere was no significant difference in operation time, intraoperative blood loss, postoperative drainage, and hospital stay between 2 groups (P>0.05). All patients were followed up 18-62 months (mean, 30.9 months). Wound complications, postoperative delirium, and Cage retropulsion occurred in 4 cases (2 cases in single-level group, 2 cases in double-level group), 1 case of single-level group, and 1 case of double-level group, respectively. The intervertebral height of fusion segments after operation significantly improved compared with preoperative ones in both groups (P<0.05). At last follow-up, the intervertebral height in double-level group was superior to which in single-level group (P<0.05). The coronal Cobb angles after operation significantly improved compared with preoperative ones (P<0.05), and no significant difference was found between 2 groups at each time point (P>0.05). The disc insertion depth showed no significant difference between different time points after operation in 2 groups (P>0.05) and between 2 groups at each time point after operation (P>0.05). Bony fusion was obtained in all patients at last follow-up. The VAS score and modified ODI after operation in both groups were superior to those before operation (P<0.05). The VAS score in double-level group was higher than that in single-level group (P<0.05) at last follow-up, and no significant difference was found in VAS score and modified ODI between 2 groups at other time points (P>0.05). According to the MacNab criteria, the excellent and good rates at last follow-up were 94.4% and 84.0% in single-level group and double-level group, respectively.ConclusionThe n-HA/PA66 Cage can effectively restore and maintain the disc height of fusion segment, normal sequence, and biomechanical stability of the spine, and gain favorable effectivenss for degenerative lumbar scoliosis. And double-level fusion is superior to single-level fusion in maintaining disc height of fusion segment.
Objective To summarize the progress of percutaneous endoscopic lumbar interbody fusion in the treatment of lumbar degenerative diseases. Methods The relevant literature about percutaneous endoscopic lumbar interbody fusion at home and abroad in recent years was reviewed, the approaches, technical characteristics, short- and long-term effectiveness, and complications of different surgical procedures were summarized. Results Percutaneous endoscopic lumbar interbody fusion is a safe and reliable treatment. At present, the main surgical methods in clinical application can be roughly summarized as percutaneous endoscopic posterior transforaminal lumbar interbody fusion (Endo-PTLIF), percutaneous endoscopic transforaminal lumbar interbody fusion (Endo-TLIF), percutaneous endoscopic oblique lumbar interbody fusion (Endo-OLIF), percutaneous endoscopic lumbar interbody fusion/Z’s percutaneous endoscopic lumbar interbody fusion (Endo-LIF/ZELIF), and unilateral biportal endoscopic transforaminal lumbar interbody fusion (UBE-TLIF). Each surgical method has its own technical characteristics and development. Conclusion Percutaneous endoscopic lumbar interbody fusion is a kind of combined technology based on the individualization of the patient’s anatomical structure and the technical differentiation of the surgeon. Surgical experience, choosing adaptive indication and operative way reasonably are the key for the success.
OBJECTIVE: To observe the early clinical results with degenerative lumbar instability treated with transpedical screw fixation and intertransverse process autogenous bone grafting. METHODS: From September 2000 to February 2002, 19 patients (5 males and 14 females) of degenerative lumbar spine instability were treated with decompression for spinal canal stenosis, transpedical screw fixation and intertransverse process autogenous bone grafting. The locations of degenerative lumbar spine instability were between L4 and L5 in 10 patients, between L3, L4 and L5 in 4 cases, between L3 and L4 in 3 cases, between L5 and S1 in 2 cases. The results were evaluated after operation. The preoperative clinical symptoms disappeared completely as excellent results, relieved obviously as good results, improved as fair results and unrelieved or worsened as poor results. RESULTS: Seventeen patients were followed up for 4-18 months with an average of 8.1 months. The results of the treatment were excellent in 12 patients, good in 4 patients and fair in 1 patient. The excellent and good rate was 94.1%. Intertransverse process arthrodesis was obtained after 6 months of operation in all cases. No loosened and broken instruments occurred. CONCLUSION: The advantages of degenerative lumbar spine instability treated with transpedical screw fixation and intertransverse are reliable fixation, high successful rate of fusion and less influence on spinal canal. The above results show satisfactory clinical outcome.
Objective To summarize the research progress of microRNA (miRNA) and its non-viral vector in intervertebral disc degeneration (IDD) and to investigate the potential of non-viral vector delivery of miRNA in clinical application. Methods The related literature about the role of miRNA in IDD and its non-viral delivery system was reviewed and analyzed. Results MiRNA can regulate the related gene expression level and further participate in the pathophysiologic process in degenerated intervertebral disc, miRNA delivered by various non-viral vectors has obtained an ideal effect in some diseases. Conclusion MiRNA plays a great role in the cellular and molecular mechanisms of IDD, as a safe and effective strategy for gene therapy, non-viral vector provides new possibilities for IDD treated with miRNA.
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.
ObjectiveTo investigate the feasibility of establishing intervertebral disc degeneration (IDD) model by using minimally invasive acupuncture and rotary-cutting. MethodsForty New Zealand white rabbits [male or female, (2.9±0.3) kg in weight] were randomly divided into control group (n=20) and experimental group (n=20). No treatment was done in the control group; percutaneous puncture was performed on L4, 5 and L5, 6 intervertebral disc by using 18G needle under C-arm X-ray monitoring for rotary-cutting of nucleus pulposus to promote degeneration of the disc in the experimental group. At 4, 8, 12, and 16 weeks after operation, general observation and MRI observation were done, and intervertebral disc degeneration was accessed based on Pfirrmann grade; the specimens were harvested for Masson staining and Safranine O staining. ResultsThe nucleus pulposus showed dark colors and reduced elasticity in the experimental group when compared with the control group. T2-weighted MRI images indicated that the disc signal intensity of control group had no obvious change at early stage, and weakened slightly at late stage; disc signal intensity of the experimental group decreased with time. According to Pfirrmann grade for disc degeneration, disc degeneration degree was significantly aggravated with time in 2 groups (P < 0.05); degeneration was significantly more severe in the experimental group than the control group at the other time points (P < 0.05) except 4 weeks (P > 0.05). Masson staining results showed that irregular arrangement of annulus with integrate structure was observed in the control group with time; the annulus of the experimental group arranged in disorder, or even disc fibrous circle rupture appeared with time. Safranin O staining showed that the nucleus pulposus cells reduced significantly in the experimental group, but did not in the control group. ConclusionMinimally invasive acupuncture and rotary-cutting could successfully establish the IDD model in rabbits.
Objective To discuss the main points of technique and the range of fusion in posterior operation of spinal stenosis associated with lumbar degenerative kyphosis (LDK). Methods The cl inical data were retrospectively analysedfrom 20 cases of spinal stenosis associated with LDK which were performed posterior operation from February 2001 to February 2008. There were 1 male and 19 females, aged 52-81 years old with an average of 64 years old. The course of disease was 6-10 years. All patients had severe low back pain. According to Frankel’s neurologic function classification, there were 18 cases of grade E and 2 cases of grade D before operation. The apex of LDK included L1 in 3 cases, L2 in 10 and L3 in 7. The operational method was decided according to different characteristics of LDK. All patients were divided into three groups. Group 1 included 6 cases of sciatica and intermittent claudication with worse physical status, the segmental decompression of spinal canal, posterior intervertebral fusion and short transpedical instrument fixation were performed. Group 2 included 8 cases whose Cobb angle of LDK was less than 20°, the segmental decompression of spinal canal, posterior intervertebral fusion and one-level or multilevel lamina osteotomy were performed, instrumentation-assisted correction was used. Group 3 included 6 cases whose Cobb angle of LDK was more than 20°, the canal decompression and one-level transvertebral wedge osteotomy were performed, instrumentation-assisted correction, intervertebral fusion and posterior-lateral fusion were used. Results Incision healedby first intention in all patients. One patient suffered from superior mesenteric artery syndrome at 6 hours after operationand healed after symptomatic management. The neurologic function was improved to grade E at 2 weeks after opeartion. All patients were followed-up 24-54 months (average 26 months). At last follow-up，the Oswestry Disabil ity Index of all patients was 30.5% ± 9.6%; showing significant difference when compared with preoperation (55.9% ± 11.8%, P lt; 0.05). The back pain scoring and leg pain scoring were 2.8 ± 1.6 and 2.4 ± 1.6, respectively according to the Numeric Rating Scale score; showing significant differences when compared with preoperation (7.5 ± 0.5 and 7.3 ± 0.7, P lt; 0.05). The Numeric Rating Scale score and Oswestry Disabil ity Index in all patients were improved obviously when compared with before operation (P lt; 0.05). During the follow-up period, there was no instrumentation failure or correction loss and the fusion rate was up to 100%. Conclusion For spinal stenosis associated with LDK patients, the most important therapic purpose is to improve cl inical symptom through reconstruction lumbar stabil ization and spinal biomechanics l ine in sagittal plane. Overall estimate of the cl inical appearance and imageology character is necessary when making decision of which segments needed to be fixation and fusion. Individual ized treatment strategy may be the best choice.
ObjectiveTo review the disc degeneration models and the current treatment status of calcitonin. MethodsRecent literature concerning the disc degeneration models and the calcitonin treatment of disc degeneration was extensively reviewed and summarized. ResultsDifferent models of disc degeneration have advantages and disadvantages; calcitonin can relieve disc degeneration at different degrees in vitro and in vivo studies. ConclusionDisc degeneration model for the study of disc degeneration mechanism offers a variety of ideas,and the results of calcitonin treatment in the disc degeneration model can provide the basis for future experiments.
ObjectiveTo observe the difference between crenel lateral interbody fusion (CLIF) and transforaminal lumbar interbody fusion (TLIF) in the treatment of degenerative lumbar spondylolisthesis (DLS) combined with lumbar spinal stenosis (LSS).MethodsThe clinical data of DLS combined with LSS patients meeting the selection criteria admitted between May 2018 and May 2019 were retrospectively analyzed. According to different surgical methods, the patients were divided into CLIF group (33 cases) and TLIF group (32 cases). There were no significant differences (P>0.05) between the two groups in gender, age, disease duration, lesion segments, lumbar bone mineral density, degree of lumbar spondylolisthesis, and preoperative visual analogue scale (VAS) score, Oswestry disability index (ODI), intervertebral space height, intervertebral foramen height, lumbar lordosis (LL), and segmental lordosis (SL). The operation time, intraoperative blood loss, and perioperative complications were recorded and compared between the two groups. Lumbar CT scan was performed at last follow-up to compare the intervertebral fusion rate between the two groups. Intervertebral space height, intervertebral foramen height, LL, and SL were measured before operation, at 2 weeks, 3 months after operation, and at last follow-up. VAS score and ODI were used to evaluate the pain and improvement of the quality of life of the patients.ResultsThere were no neurological and vascular complications in the two groups. The operation time and intraoperative blood loss in CLIF group were significantly less than those in TLIF group (P<0.05). Patients in both groups were followed up for a median time of 18 months. All the incisions healed by first intention except 1 incision in TLIF group because of poor blood glucose control. No complications such as bedsore, falling pneumonia, and deep venous thrombosis were found in both groups. At last follow-up, the intervertebral fusion rates in CLIF and TLIF group were 90.91% (30/33) and 93.75% (30/32), respectively, showing no significant difference (χ2=0.185, P=0.667). The VAS score, ODI, intervertebral space height, intervertebral foramen height, LL, and SL were significantly improved in both groups at each time point after operation (P<0.05). Except that VAS score in CLIF group was significantly lower than that in TLIF group at 2 weeks after operation (Z=−4.303, P=0.000), there were no significant differences in VAS score and ODI between the two groups at other time points (P>0.05). The intervertebral space height, intervertebral foramen height, LL, and SL in CLIF group were significantly higher than those in TLIF group at each time point after operation, and the differences were significant (P<0.05).ConclusionCLIF in the treatment of DLS combined with LSS can achieve the similar effectiveness with traditional TLIF, and has such advantages as minimal invasion and faster recovery.