Enhanced pyroptosis induction with pore-forming gene delivery for osteosarcoma microenvironment reshaping.

Osteosarcoma is the most common malignant bone tumor without efficient management for improving 5-year event-free survival. Immunotherapy is also limited due to its highly immunosuppressive tumor microenvironment (TME). Pore-forming gasdermins (GSDMs)-mediated pyroptosis has gained increasing concern in reshaping TME, however, the expressions and relationships of GSDMs with osteosarcoma remain unclear. Herein, gasdermin E (GSDME) expression is found to be positively correlated with the prognosis and immune infiltration of osteosarcoma patients, and low GSDME expression was observed. A vector termed as LPAD contains abundant hydroxyl groups for hydrating layer formation was then prepared to deliver the GSDME gene to upregulate protein expression in osteosarcoma for efficient TME reshaping via enhanced pyroptosis induction. Atomistic molecular dynamics simulations analysis proved that the hydroxyl groups increased LPAD hydration abilities by enhancing coulombic interaction. The upregulated GSDME expression together with cleaved caspase-3 provided impressive pyroptosis induction. The pyroptosis further initiated proinflammatory cytokines release, increased immune cell infiltration, activated adaptive immune responses and create a favorable immunogenic hot TME. The study not only confirms the role of GSDME in the immune infiltration and prognosis of osteosarcoma, but also provides a promising strategy for the inhibition of osteosarcoma by pore-forming GSDME gene delivery induced enhanced pyroptosis to reshape the TME of osteosarcoma.

Satisfactory immediate spontaneous correction may not mean satisfactory final results for moderate TL/L curves after selective thoracic fusion in AIS patients.

BACKGROUND: Few studies have focused on the chronic spontaneous behavior of the unfused TL/L curve during follow-up. The purpose of the present study was to explore the behavior of the unfused TL/L curve during a long-term follow-up to identify the risk factors for correction loss. METHODS: Sixty-four age-matched female AIS patients undergoing selective thoracic fusion were enrolled. Patients were divided into 2 groups according to whether there was correction loss. Risk factors for correction loss of the unfused TL/L curves were analyzed. The relationship and difference between the immediate postoperative thoracic and TL/L Cobb angles were explored. RESULTS: The TL/L Cobb angle was 28.17° before surgery, 8.60° after surgery, and 10.74° at the final follow-up, with a correction loss of 2.14°. Each subgroup contained 32 cases. A smaller postoperative TL/L Cobb angle was the only risk factor that was independently associated with TL/L correction loss. In the LOSS group, there was a significant difference and no correlation between the immediate postoperative TL/L and the thoracic Cobb angle. In the NO-LOSS group, there was a moderate correlation and no difference between them. CONCLUSION: A smaller immediate postoperative TL/L Cobb angle may have been associated with TL/L correction loss during the long-term follow-up. Thus, good immediate postoperative spontaneous correction may not mean a satisfactory outcome at the final follow-up after STF. Mismatch between thoracic and TL/L Cobb angles immediately after surgery may also be related to correction loss of the unfused TL/L curves. Close attention should be paid in case of deterioration.

Clinical and Radiographic Comparison of Oblique Lateral Lumbar Interbody Fusion and Minimally Invasive Transforaminal Lumbar Interbody Fusion in Patients with L4/5 grade-1 Degenerative Spondylolisthesis.

OBJECTIVES: To compare the clinical and radiographic outcomes of oblique lateral lumbar interbody fusion and minimally invasive transforaminal lumbar interbody fusion in patients with grade-1 L4/5 degenerative spondylolisthesis. METHODS: Based on the inclusion and exclusion criteria, the comparative analysis included consecutive patients with grade-1 degenerative spondylolisthesis who underwent oblique LIF (OLIF, n = 36) or minimally invasive transforaminal LIF (MI-TLIF, n = 45) at the Department of Spine Surgery, Beijing Jishuitan Hospital from January 2016 to August 2017. Patient satisfaction Japanese Orthopaedic Association score, visual analog scale (VAS) scores for back and leg pain, Oswestry disability index (ODI), radiographic outcomes including anterior/posterior disc heights (ADH/PDH), foraminal height (FH), foraminal width (FW), cage subsidence, cage retropulsion, and fusion rate were assessed during a 2-year follow-up. Continuous data are presented as mean ± standard deviation and were compared between groups using the independent sample t-test. Categorical data are presented as n (%) and were compared between groups using the Pearson chi-squared test or Fisher's exact test. Repetitive measurement and analysis of variance was employed in the analysis of ODI, back pain VAS score, and leg pain VAS score. Statistical significance was defined as p < 0.05. RESULTS: The OLIF and MI-TLIF groups comprised 36 patients (age, 52.1 ± 7.2 years; 27 women) and 45 patients (age, 48.4 ± 14.4 years; 24 women), respectively. Satisfaction rates at 2 years post procedure exceeded 90% in both groups. The OLIF group had less intraoperative blood loss (140 ± 36 vs 233 ± 62 mL), lower back pain VAS score (2.42 ± 0.81 vs 3.38 ± 0.47), and ODI score (20.47 ± 2.53 vs 27.31 ± 3.71) at 3 months follow-up (with trends toward lower values at 2 years follow-up), but higher leg pain VAS scores at all postoperative time points than the MI-TLIF group (all p < 0.001). ADH, PDH, FD, and FW improved in both groups post-surgery. At the 2 year follow-up, the OLIF group had a higher rate of Bridwell grade-I fusion (100% vs 88.9%, p = 0.046) and lower incidences of cage subsidence (8.33% vs 46.67%, p < 0.001) and retropulsion (0% vs 6.67%, p = 0.046) than the MI-TLIF group. CONCLUSIONS: In patients with grade-I spondylolisthesis, OLIF was associated with lower blood loss and greater improvements in VAS for back pain and ODI and radiologic outcomes than MI-TLIF. The OLIF is more suitable for these patients with low back pain as the main symptoms are accompanied by mild or no leg symptoms before operation.

Diabetes and intervertebral disc degeneration: A Mendelian randomization study.

Introduction: Intervertebral disc degeneration (IVDD) is an important contributor of low back pain, which represents one of the most disabling symptoms within the adult population. Recently, increasing evidence suggests the potential association between Type 2 diabetes mellitus (T2DM) and IVDD. However, the causal relationship between these two common diseases remains unclear. Methods: We conducted a two-sample Mendelian randomization (MR) analysis to assess the causal association between T2DM and IVDD. Sensitivity analysis was performed to test for heterogeneity and horizontal pleiotropy. Multivariable MR was also conducted to adjust for the effect of BMI on IVDD. Results: A total of 128 independent single-nucleotide polymorphisms (SNPs) that were significantly associated with T2DM were selected as instrumental variables in univariable MR analysis. Our results showed that patients with T2DM had a higher risk of developing IVDD (OR, 1.069; 95% CI, 1.026-1.115; p = 0.002). The relationship remained stable in sensitive analysis including multivariable MR, which implicated the direct causal effect of T2DM on IVDD (OR, 1.080; 95% CI, 1.041-1.121; p < 0.001) after adjusting for BMI. Conclusions: MR analysis indicated a causal effect of T2DM on IVDD, and the effect persisted even when we accounted for the impact of BMI.

Where should Scoliometer and EOS Imaging be Applied when Evaluating Spinal Rotation in Adolescent Idiopathic Scoliosis -A Preliminary Study with Reference to CT Images.

STUDY DESIGN: Retrospective study. OBJECTIVE: Our purpose was to evaluate spinal rotation measurement by scoliometer or EOS Imagings with reference to that by CT images, and to clarify their applicability in clinical practice. METHODS: Patients with adolescent idiopathic scoliosis (AIS) who were indicated for surgery were enrolled and the informed consents were obtained. The angle of trunk rotation (ATR) was measured by the scoliometer. Apical vertebral rotation (AVR) was measured with EOS Imaging and CT images. Paired T tests were used to compare the measurements between ATR or AVR-EOS and AVR-CT. Pearson correlation analysis was performed to explore the relationship between ATR or AVR-EOS and AVR-CT. Then subgroup analysis was performed. RESULTS: Forty-seven consecutive AIS patients with 62 curves were identified. In the whole group, the ATR, as well as AVR-EOS, was significantly smaller than the AVR-CT. Both ATR and AVR-EOS correlated with AVR-CT, although AVR-EOS correlated better. In thoracic group, there was no significant difference between ATR and AVR-CT (P = .236). A significant correlation was found between ATR and AVR-CT(r = .574, P < .001). In TL/L group, no significant difference was noted between AVR-EOS and AVR-CT (P = .414), and a significant correlation was found between AVR-EOS and AVR-CT(r = .824, P < .001). CONCLUSION: ATR by scoliometer is numerically similar to AVR by CT and may evaluate the spinal rotation more appropriately in thoracic spine. AVR by EOS is numerically similar to AVR by CT and may be more applicable in TL/L spine. Appropriate methods could be selected according to the location of the curve.

Longitudinal Spinous-Splitting Laminoplasty with Coral Bone for the Treatment of Cervical Adjacent Segment Degenerative Disease: A 5-Year Follow-up Study.

This study was designed to analyze the causes of cervical adjacent segment degenerative disease (ASDis), evaluate the surgical outcomes of longitudinal spinous-splitting laminoplasty with coral bone (SLAC) during cervical reoperation, and accumulate data on reoperation with SLAC in a primary hospital. Based on the inclusion and exclusion criteria, we conducted a retrospective study involving 52 patients who underwent cervical reoperation for ASDis using SLAC at the spinal surgery department of the Beijing Jishuitan Hospital from 1998 to 2014. Among them, 39 were treated with anterior cervical fusion and internal fixation during the first operation (anterior cervical corpectomy with fusion [ACCF], n = 24; anterior cervical discectomy and fusion [ACDF], n = 11; and cervical disc arthroplasty [CDA], n = 4). Outcomes were the Japanese Orthopaedic Association (JOA) score, neck disability index (NDI) score, upper limb/neck and shoulder evaluated using a visual analogue scale (VAS), and rates of ASDis. In patients who underwent an anterior cervical approach in the first instance, the incidence of ASDis was significantly higher in the C3/4 gap than in the other gaps. In the ACCF group, the lateral radiograph of the cervical spine revealed that the distance between the anterior cervical plate and the adjacent segment disc was <5 mm in 15 (62.5%) cases and five (12.8%) cases, respectively, the internal fixation screws broke into the annulus of the adjacent segment. After the first SLAC, ASDis developed at C2/3 and C3/4 in four (30.8%) and eight (61.5%) cases, respectively. After reoperation, all cases were followed up for >5 (average, 6.2) years. The pre-reoperation and last follow-up values were as follows: mean Japanese Orthopaedic Association score, 10.2 ± 1.5 vs 15.5 ± 0.7 (P = 0.03); neck disability index, 26.2 vs 13.6 points (P = 0.01); upper-limb visual analog scale (VAS) score, 6.1 vs 2.6 points (P = 0.04); and neck and shoulder VAS score, 6.6 vs 2.1 points (P = 0.03). SLAC is a simple technique in which the local anatomy is clearly visible and satisfactory clinical outcomes are obtained.

Comparison of Outcomes between Robot-Assisted Minimally Invasive Transforaminal Lumbar Interbody Fusion and Oblique Lumbar Interbody Fusion in Single-Level Lumbar Spondylolisthesis.

OBJECTIVE: To compare the safety and effectiveness of robot-assisted minimally invasive transforaminal lumbar interbody fusion (Mis-TLIF) and oblique lumbar interbody fusion (OLIF) for the treatment of single-level lumbar degenerative spondylolisthesis (LDS). METHODS: This is a retrospective study. Between April 2018 and April 2020, a total of 61 patients with single-level lumbar degenerative spondylolisthesis and treated with robot-assisted OLIF (28 cases, 16 females, 12 males, mean age 50.4 years) or robot-assisted Mis-TLIF (33 cases, 18 females, 15 males, mean age 53.6 years) were enrolled and evaluated. All the pedicle screws were implanted percutaneously assisted by the TiRobot system. Surgical data included the operation time, blood loss, and length of postoperative hospital stay. The clinical and functional outcomes included Oswestry Disability Index (ODI), Visual Analog scores (VAS) for back and leg pain, complication, and patient's satisfaction. Radiographic outcomes include pedicle screw accuracy, fusion status, and disc height. These data were collected before surgery, at 1 week, 3 months, 6 months, and 12 months postoperatively. RESULTS: There were no significantly different results in preoperative measurement between the two groups. There was significantly less blood loss (142.4 ± 89.4 vs 291.5 ± 72.3 mL, P < 0.01), shorter hospital stays (3.2 ± 1.8 vs 4.2 ± 2.5 days, P < 0.01), and longer operative time (164.9 ± 56.0 vs 121.5 ± 48.2 min, P < 0.01) in OLIF group compared with Mis-TLIF group. The postoperative VAS scores and ODI scores in both groups were significantly improved compared with preoperative data (P < 0.05). VAS scores for back pain were significantly lower in OLIF group than Mis-TLIF group at 1 week (2.8 ± 1.2 vs 3.5 ± 1.6, P < 0.05) and 3 months postoperatively (1.6 ± 1.0 vs 2.1 ± 1.1, P < 0.05), but there was no significant difference at further follow-ups. ODI score was also significantly lower in OLIF group than Mis-TLIF group at 3 months postoperatively (22.3 ± 10.0 vs 26.1 ± 12.8, P < 0.05). There was no significant difference in the proportion of clinically acceptable screws between the two groups (97.3% vs 96.2%, P = 0.90). At 1 year, the OLIF group had a higher interbody fusion rate compared with Mis-TLIF group (96.0% vs 87%, P < 0.01). Disc height was significantly higher in the OLIF group than Mis-TLIF group (12.4 ± 3.2 vs 11.2 ± 1.3 mm, P < 0.01). Satisfaction rates at 1 year exceeded 90% in both groups and there was no significant difference (92.6% for OLIF vs 91.2% for Mis-TLIF, P = 0.263). CONCLUSION: Robot-assisted OLIF and Mis-TLIF both have similar good clinical outcomes, but OLIF has the additional benefits of less blood loss, less postoperative hospital stays, higher disc height, and higher fusion rates. Robots are an effective tool for minimally invasive spine surgery.

Biomechanical Evaluation of the Transcortical and Transpedicular Trajectories for Pedicle Screw Insertion in Thoracolumbar Fracture Fixation for Ankylosing Spondylitis.

Background: Ankylosing spondylitis (AS) is a chronic disorder characterized by an imbalance between bone formation and resorption. Spinal fractures often occur after minor trauma in patients with AS. For thoracolumbar fractures, transpedicular screw (TPS) fixation through the posterior approach has been suggested. The cortical bone trajectory (CBT) technique has also been used to prevent screw pull-out in patients with poor bone quality. The aim of current study was to assess the biomechanical characteristics of the TPS and CBT technique in thoracolumbar AS fracture fixation by finite element analysis. Methods: The three-dimensional finite element models of the AS spine were created. The CBT and TPS methods of screw insertion were used in AS spinal fracture models. An intact AS spine model was considered the control. An axial force and torsion in rotation, flexion/extension and lateral flexion were applied in all models in CBT, TPS, and control groups. Results: The AS spine showed similar construct stiffness after posterior fixation by CBT and TPS techniques under axial, rotational, and flexion/extension loading conditions. The TPS technique showed better intact stability under all loading conditions. Similarly, the TPS technique provided superior fracture regional stability against axial and rotational loads than did the CBT technique. The maximum von Mises stresses were 1714.4 ± 129.8 MPa and 1208.7 ± 107.3 MPa (p < 0.001), which occurred in the CBT and TPS groups under compressive loading. Conclusions: The TPS technique provides better biomechanical strength under axial, rotational, flexion/extension, and lateral flexion loading than does the CBT technique. Compared with CBT, TPS is more effective in maintaining the stability of AS thoracolumbar fractures from a finite element analysis perspective.

Standalone oblique lateral interbody fusion vs. combined with percutaneous pedicle screw in spondylolisthesis.

BACKGROUND: To compare standalone oblique lateral interbody fusion (OLIF) vs. OLIF combined with posterior bilateral percutaneous pedicle screw fixation (OLIF combined) for the treatment of lumbar spondylolisthesis. METHODS: This was a retrospective study of patients who underwent standalone OLIF or combined OLIF between 07/2014 and 08/2017 at two hospitals in China. Direct decompressions were not performed. Visual analog scale (VAS), Oswestry Disability Index (ODI), satisfaction rate, anterior/posterior disc heights (DH), foraminal height (FH), foraminal width (FW), cage subsidence, cage retropulsion, fusion rate, and complications were analyzed. All imaging examinations were read independently by two physicians and the mean measurements were used for analysis. RESULTS: A total of 73 patients were included: 32 with standalone OLIF and 41 with combined OLIF. The total complication rate was 25.0% with standalone OLIF and 26.8% with combined OLIF. There were no differences in VAS and ODI scores by 2 years of follow-up, but the scores were better with standalone OLIF at 1 week and 3 months (P < 0.05). PDH and FW was smaller in the combined OLIF group compared with the standalone OLIF group before and after surgery (all P < 0.05). There were significant differences in FH before surgery and at 1 week and 3 months between the two groups (all P < 0.05), but the difference disappeared by 2 years (P = 0.111). Cage subsidence occurred in 7.3% (3/41) and 7.3% (3/41) of the patients at 3 and 24 months, respectively, in the combined OLIF group, compared with 6.3% (2/32) and 15.6% (5/32), respectively, in the standalone OLIF group at the same time points (P = 0.287). There was no cage retropulsion in both groups at 2 years. The fusion rate was 85.4%(35/41) in the combined OLIF group and 84.4% (27/32) in the standalone OLIF group at 3 months(P = 0.669). At 24 months, the fusion rate was 100.0% in the combined OLIF group and 93.8% (30/32) in the standalone OLIF group (P = 0.066). CONCLUSION: Standalone OLIF may achieve equivalent clinical and radiological outcomes than OLIF combined with fixation for spondylolisthesis. The rate of complications was similar between the two groups. Patients who are osteoporotic might be better undergoing combined rather than standalone OLIF. The possibilty of proof lies within a future prospective study, preferably an RCT.

Quantitative analysis of paraspinal muscle atrophy after oblique lateral interbody fusion alone vs. combined with percutaneous pedicle screw fixation in patients with spondylolisthesis.

BACKGROUND: There is no available literature for comparison on muscle atrophy between the "stand-alone" oblique lateral interbody fusion (OLIF) and regular OLIF (i.e., combined with percutaneous pedicle screws fixation (PPSF) in patients with spondylolisthesis). This study aimed to identify changes in back muscle atrophy between the two surgeries. METHODS: This was a retrospective cohort study of patients who underwent OLIF or OLIF+PPSF at Beijing Jishuitan Hospital and Shanghai ChangZheng Hospital between 07/2014 and 10/2017. Computed tomography (CT) was used to measure functional cross-sectional area (FCSA) and fat infiltration percentage (FIP) of the multifidus and erector spinae before and 24 months after surgery. RESULT: There were no differences in FCSA and FIP between OLIF (n = 32) and OLIF+PPSF (n = 41) groups before surgery. In the OLIF group, the multifidus and erector spinae FCSA and FIP did not change at 24 months (FCSA: multifidus: from 8.59 ± 1.76 to 9.39 ± 1.74 cm2, P = 0.072; erector spinae: from 13.32 ± 1.59 to 13.55 ± 1.31 cm2, P = 0.533) (FIP: multifidus: from 15.91 ± 5.30% to 14.38 ± 3.21%, P = 0.721; erector spinae: from 11.63 ± 3.05% to 11.22 ± 3.12%, P = 0.578). In the OLIF+PPSF group, the multifidus and erector spinae FCSA decreased (multifidus: from 7.72 ± 2.69 to 5.67 ± 1.71 cm2, P < 0.001; erector spinae: from 12.60 ± 2.04 to 10.15 ± 1.82 cm2, P < 0.001), while the FIP increased (multifidus: from 16.13 ± 7.01% to 49.38 ± 20.54%, P < 0.001; erector spinae: from 11.93 ± 3.22% to 22.60 ± 4.99%, P < 0.001). The differences of FCSA and FIP between the two groups at 24 months were significant (all P < 0.001). The patients in the standalone OLIF group had better VAS back pain, and JOA scores than the patients in the OLIF combined group (all P < 0.05) at 1 week and 3 months after surgery. There were two cases (4.9%) of adjacent segment degeneration in the OLIF combined group, while there was no case in the OLIF alone group. CONCLUSIONS: Standalone OLIF had better clinical outcomes at 1 week and 3 months than OLIF+PPSF in patients with spondylolisthesis. OLIF may not result in paraspinal muscle atrophy at 24 months after surgery.

Secreted Factors From Intervertebral Disc Cells and Infiltrating Macrophages Promote Degenerated Intervertebral Disc Catabolism.

STUDY DESIGN: Rat nucleus pulposus (NP) cells or annulus fibrosus (AF) cells were stimulated with conditioned media of RAW 264.7 macrophages and vice versa under healthy culture conditions and in the presence of pro-inflammatory mediators. The gene expression of pro-inflammatory mediators, extracellular matrix (ECM)-modifying enzymes, and chemokines, which play important roles in intervertebral disc degeneration (IDD), was determined. OBJECTIVE: To test whether the interaction between native disc cells and infiltrating macrophages accelerates inflammation state, disrupts matrix homeostasis, and promotes inflammatory cells infiltration. SUMMARY OF BACKGROUND DATA: With macrophages infiltration, the disc resident cells would be inevitably exposed to macrophages. Macrophages have been shown to play pro-inflammatory role in the cellular interactions with disc cells under healthy culture conditions. However, the biologic interactions between macrophages and disc cells under degenerated disc inflammatory environment remain unknown. METHODS: Murine Macrophages RAW 264.7 were cultured in the conditioned media of Rat AF or NP cells culture in the presence or absence of IL-1ß stimulation. Similarly, Rat AF or NP cells were also cultured in the conditioned media of Murine Macrophages RAW 264.7 culture in the presence or absence of IFN-γ stimulation. The mRNA levels difference of pro-inflammatory genes, catabolic genes and chemokines genes for AF cells, NP cells and Macrophages RAW 264.7 were analyzed by qRT-PCR, respectively. RESULTS: Compared with serum-free media exposure, RAW 264.7 macrophages exposed to AF or NP cells conditioned media selectively modestly upregulated mRNA levels of the aforementioned cytokines. Exposure of RAW 264.7 macrophages to conditioned media from AF or NP cells with IL-1ß stimulation dramatically increased mRNA levels of all the investigated cytokines. Similarly, compared with serum-free media exposure, AF or NP cells exposed to RAW 264.7 macrophages conditioned media selectively modestly upregulated mRNA levels of the aforementioned cytokines. Exposure of AF or NP cells to conditioned media from RAW 264.7 macrophages with IFN-γ stimulation dramatically increased mRNA levels of all the investigated cytokines. CONCLUSION: The biologic interactions between infiltrating macrophages and native disc cells under degenerated disc inflammatory environment lead to an increasingly severe inflammatory conditions, which may be a self-stimulated process from the macrophages infiltration occurrenceLevel of Evidence: 5.

[Cause analysis of back pain after vertebroplasty or kyphoplasty].

OBJECTIVE: To analyze the causes of back pain after vertebroplasty or kyphoplasty. METHODS: 1 863 patients with back pain who underwent vertebroplasty or kyphoplasty from Jan. 2004 to Dec. 2014 in Department of Spine Surgery, Beijing Jishuitan Hospital, were followed up for least 3 months after surgery with their back pain evaluated by VAS scores. Related information including preoperative, operative and postoperative factors was gathered. The VAS at last follow-up over 5 or the symptom relief less than 50% was considered as non-relieved, the correlations between the symptom relief and potential influencing factors were analyzed by the Spearman correlation. RESULTS: Symptom relief was only correlated with postoperative complications (P<0.001). The back pain in 283 cases (15.2%) had not been relieved. Among them, 87 cases got fractures in other vertebrae (adjacent vertebrae in 67 cases, non-adjacent vertebrae in 20 cases); 3 cases got nonunion in the operated vertebrae; 1 case got postoperative infection; 6 cases got misdiagnosis and the other 186 cases had no known causes for the unrelieved pain. CONCLUSIONS: Postoperative back pain after vertebroplasty or kyphoplasty may have variable causes, and some of which have not been entirely understood. If the symptoms are not alleviated or even aggravate after active symptomatic treatment and anti-osteoporotic therapy, underlying causes such as re-fracture, nonunion, infection, pathologic fracture should be sought out.

COX2 is involved in hypoxia-induced TNF-α expression in osteoblast.

Bone regeneration involves a series of events in a coordinated manner, including recruitment of mesenchymal stem cells, induction of immune response, inflammatory activity and vascular ingrowth. The microenvironment of bone regeneration is hypoxic. Low oxygen tension (hypoxia) promotes the upregulation of several signaling molecules. The primary mediating factor is the hypoxia-inducible factor-1 (HIF-1). Hypoxia stimulates the expression of a variety of cytokines from inflammatory cells, fibroblasts, endothelial cells, and osteoblasts. TNF-α is a key proinflammatory cytokine. The molecular events involved in osteoblast dysfunction under hypoxia are not fully understood. This study determined the effects of hypoxia on TNF-α in osteoblasts, and molecular mechanisms were explored. We observed that hypoxia induced TNF-α expression in a time-dependent manner in osteoblasts. Experiments using a potent HIF-1α activator DFO demonstrated that hypoxia-induced TNF-α was mediated by HIF-1-α. In addition, this study showed that hypoxia activated cyclooxygenase-2 (COX2) expression along with TNF-α. Inhibition experiments using COX2 inhibitor N398 indicated that COX2 was involved in hypoxia-mediated TNF-α expression, and this observation was further confirmed by Small interfering RNA against COX2. On the other hand, TNF-α didn't lead to the activation of COX2 expression. We conclude that COX2 is involved in hypoxia-induced TNF-α expression in osteoblast.

The effect of lateral wall perforation on screw pull-out strength: a cadaveric study.

BACKGROUND: Lateral pedicle wall perforations occur frequently during pedicle screw insertion. Although it is known that such an occurrence decreases the screw pull-out strength, the effect has not been quantified biomechanically. MATERIALS AND METHODS: Twenty fresh cadaveric lumbar vertebrae were harvested, and the bone mineral density (BMD) of each was evaluated with dual-energy radiography absorptiometry (DEXA). Twenty matched, 6.5-mm pedicle screws were inserted in two different manners in two groups, the control group and the experimental group. In the control group, the pedicle screw was inserted in a standard fashion taking adequate precaution to ensure there was no perforation of the wall. In the experimental group, the pedicle screw was inserted such that its trajectory perforated the lateral wall. Group assignments were done randomly, and the maximal fixation strength was recorded for each screw pull-out test with a material-testing system (MTS 858 II). RESULTS: The average BMD for both groups was 0.850 g/cm(2) (0.788-0.912 g/cm(2)). The average (and standard deviation) maximal pull-out forces were 1,015.8 ± 249.40 N for the experimental group and 1,326.0 ± 320.50 N for the control group. According to a paired t-test, the difference between the two groups was statistically significant (P < 0.001). CONCLUSION: The results of this study confirm that the maximal pull-out strength of pedicle screws decreases by approximately 23.4% when the lateral wall is perforated.

Clinical evaluation of the polymethylmethacrylate-augmented thoracic and lumbar pedicle screw fixation guided by the three-dimensional navigation for the osteoporosis patients.

PURPOSE: To evaluate the safety and efficacy of three-dimensional (3D) navigation-guided polymethylmethacrylate (PMMA)-augmented thoracic and lumbar pedicle screw fixation for the osteoporotic patients. METHODS: 27 consecutive osteoporosis patients with a variety of spinal disorders who underwent 3D navigation-guided PMMA-augmented pedicle screw fixation were evaluated clinically and radiologically in the perioperative and 1-year follow-up period. The improvement of Japanese Orthopaedic Association (JOA) scores was analyzed. PMMA leakage and other complications were inspected intraoperatively and postoperatively. Screw loosening and bone fusion were evaluated radiographically during follow-up. RESULTS: 8 patients had thoracic and lumbar fractures; 18 patients had degenerative spinal disorders; one patient had revision surgery. One patient died of postoperative pneumonia. Each of the other 26 patients was followed up regularly at 3, 6, 12 and 18 months postoperatively. The mean therapeutic improvement rate is 39.6% evaluated by JOA scores. 2 patients (7.4%) had leakage into the spinal canal in front of the posterior longitudinal ligament and two patients (7.4%) had leakage into the prevertebral soft tissue inspected by the postoperative CT scans. No pedicle cortex breach and cement leakage surrounding pedicle cortex were observed. None of patients complained of dyspnoea and showed evidence of pulmonary embolism. Bone fusions were found in 20 patients (bony fusion rate 76.9%) at the 12-month follow-up and no screw loosening occurs. CONCLUSION: The results show favorable outcome using 3D navigation-guided PMMA-augmented thoracic and lumbar pedicle screw fixation for the osteoporosis patients both clinically and radiologically.

HIF-1α inhibits Wnt signaling pathway by activating Sost expression in osteoblasts.

The nature of the cellular and molecular mechanisms for the transition of avascular cartilage replacement with bone during endochondral ossification remains poorly understood. One of the driving forces is hypoxia. As a master regulator of hypoxia, hypoxia-inducible factor-1α (HIF-1α) has been reported to couple angiogenesis to osteogenesis. Our recent study has demonstrated that osteoblast growth is inhibited under hypoxia and that HIF-1α cooperates with Osterix (Osx) to inhibit Wnt pathway. However, molecular mechanisms for inhibitory effects of HIF-1α on Wnt pathway are not well understood. In this study, our quantitative RT-PCR results revealed that the expression of a Wnt antagonist Sclerostin (Sost) was upregulated in osteoblasts during hypoxia while HIF-1α was upregulated. Treatment of desferrioxamine (DFO), a HIF-1α activator, led to further increase of Sost expression, suggesting that HIF-1α may activate Sost expression. The regulation of Sost gene expression by HIF-1α was then investigated. We performed loss-of-function experiments to examine Sost expression by using siRNA approach against HIF-1α, and found that the inhibition of HIF-1α by siRNA in osteoblasts led to the decrease of Sost expression. To address transcriptional regulation of Sost gene by HIF-1α, transient transfection assay was performed and showed that HIF-1α activated Sost-1 kb promoter reporter activity in a dose-dependent manner. To narrow down the minimal region of Sost promoter activated by HIF-1α, we generated a series of deletion mutants of Sost constructs. It was demonstrated that Sost-260 was the minimal region of Sost promoter for HIF-1α activation and that Sost-106 construct, which lack hypoxia response element, abolished HIF-1α-mediated Sost reporter activation. Gel shift assay showed that HIF-1 bound to the promoter sequence of Sost directly. These findings support our hypothesis that HIF-1α activates Sost expression. This study provides a novel molecular mechanism through which HIF-1α inhibits Wnt signaling in osteoblasts.

Occipital-C2 transarticular fixation for occipitocervical instability associated with occipitalization of the atlas in patients with klippel-feil syndrome, using intraoperative 3-dimensional navigation system.

STUDY DESIGN: Retrospective case series. OBJECTIVE: The aim of this study was to describe the clinical outcomes of cervical reduction and occipital-C2 transarticular (OCTA) fixation with an assistance of intraoperative 3-dimensional navigation system (ITNS) during the treatment of reducible occipitocervical instability (OCI) in patients with Klippel-Feil syndrome (KFS) with occipitalization of the atlas and fusion of C2-C3. SUMMARY OF BACKGROUND DATA: Patients with KFS have congenital fusions of at least 2 cervical segments and may gradually develop symptoms at the hypermobile articulations adjacent to the cervical synostosis. This is particularly common in patients with KFS with occipitalization of the atlas and C2-C3 fusion. These patients may be at risk for instability and neurological complications of the occipitocervical junction that require occipitocervical reconstruction and fusion. Numerous treatment techniques are available for this pathological condition. However, there has yet to be a study of reducible OCI, showing successful treatment with intraoperative reduction and posterior OCTA fixation using ITNS. METHODS: From 2006 to 2011, 9 patients with KFS with reducible OCI attributed to occipitalization of the atlas and C2-C3 fusion were surgically treated. After a limited foramen magnum decompression, reduction of the OCI was conducted by intraoperative cervical traction and extension, followed by OCTA fixation using a direct posterior approach and with the assistance of ITNS. The follow-up period ranged from 6 to 60 months (mean, 31 mo). RESULTS: Good decompression and bone fusion were achieved in all the patients. The clinical symptoms had improved for all patients. There were no intraoperative or postoperative complications. CONCLUSION: In patients with KFS with occipitalization of the atlas and C2-C3 fusion, manual cervical traction in tandem with cervical extension, followed by posterior OCTA fixation and fusion provides a safe, effective treatment of OCI and ventral brainstem impingement. IFTN is a feasible tool for monitoring cervical reduction and OCTA screw insertion in patients with KFS with this pathological condition.

Synergistic inhibition of Wnt pathway by HIF-1α and osteoblast-specific transcription factor osterix (Osx) in osteoblasts.

Osterix (Osx) is an osteoblast-specific transcription factor required for osteoblast differentiation. Inhibition of Wnt pathway by Osx highlights the potential for feedback control mechanisms involved in bone formation. Hypoxia-inducible factor-1α (HIF-1α) is a master regulator of hypoxia. HIF-1α has been reported to couple angiogenesis to osteogenesis. Our recent study has demonstrated that Osx and HIF-1α cooperatively regulate VEGF expression in osteoblasts. Effects of hypoxia/HIF-1α on osteoblast proliferation and related mechanisms are not well understood. In this study, osteoblast growth under hypoxia was examined. We observed that osteoblast growth was inhibited under hypoxia. To explore possible mechanisms for hypoxia/HIF-1α to inhibit osteoblast proliferation, we tested the effect of hypoxia/HIF-1α on Wnt pathway. Quantitative RT-PCR results revealed that Wnt target genes such as cyclin D1 and c-Myc were downregulated under hypoxia while HIF-1α was upregulated. Treatment of desferrioxamine, a HIF-1α activator, led to further downregulation of expressions of cyclin D1 and c-Myc in osteoblasts. On the contrary, the inhibition of HIF-1α by siRNA in osteoblasts led to the expression increase of cyclin D1 and c-Myc. These data suggest that HIF-1α inhibits Wnt pathway in osteoblasts. To examine the effect of HIF-1α on Wnt pathway, HIF-1α was cotransfected with ß-catenin along with Topflash reporter in transient transfection assay. Our results showed that HIF-1α inhibited ß-catenin-induced Topflash reporter activity. Interestingly, a synergistic interplay was observed between Osx and HIF-1α in the inhibition of ß-catenin-induced Topflash expression. Our findings indicate that Osx and HIF-1α cooperatively inhibit Wnt pathway. This study revealed additional new information of the cooperation between HIF-1α and Osx in osteoblasts.

[One-stage posterior approach spinal reconstruction for severe lumbar burst fracture].

OBJECTIVE: To evaluate the clinical therapeutic outcomes for severe burst fracture of lumbar vertebrae via a posterior approach for one-stage spinal "three-column" reconstruction. METHODS: An retrospective analysis of data was performed for 21 patients with severe burst fracture of lumbar vertebrae undergoing a posterior approach for one-stage spinal "three-column" reconstruction from 2005 to 2010. It was compared with previous 15 cases undergoing a staged anteroposterior approach. The operative duration, volume of blood loss, pre- and post-operative imaging measurements of kyphosis and vertebral height and nerve function recovery were evaluated. RESULTS: The values of operative duration and volume of blood loss in the one-stage posterior approach group were significantly less than those of the two-stage anteroposterior approach group [(263 ± 72) min vs (439 ± 75) min, t = -5.303, P < 0.01; (1143 ± 296) ml vs (1471 ± 399) ml, t = -2.169, P = 0.042)]. Statistical significance existed in postoperative kyphosis between two groups [(0.5 ± 2.0)° vs (3.9 ± 2.6)°, t = -3.336, P = 0.003]. Vertebral height had no statistical significance pre- and post-operatively between two groups while restoration of vertebral height did [(0.47 ± 0.19) mm vs (0.26 ± 0.15) mm, t = 2.669, P = 0.015]. CONCLUSION: Posterior approach for one-stage vertebral resection, mesh implantation, pedicle screws and rod internal fixation for reconstructing spinal "three-column" structures offers excellent feasibility and safety. And it may avoid complications associated with an anteroposterior approach for two-stage procedures. The median length of hospital stay is also reduced.