Lifeng Lao

Effect of disc degeneration on lumbar segmental mobility analyzed by kinetic magnetic resonance imaging.

Study Design. Retrospective radiographical study. Objective. To define the relationship between the grade of disc degeneration and the motion of the lumbar spine by using kinetic magnetic resonance imaging. Summary of Background Data. Disc degeneration is common after middle age. Lumbar instability has generally been recognized as a potential risk factor of low back pain. However, correlations between the grade of disc degeneration and the motion of the lumbar spine need more investigation. Methods. Kinetic magnetic resonance imaging was performed in 162 patients with symptomatic low back pain without prior history of surgery. The lumbar intervertebral discs were graded by spine surgeons according to the degenerative grading system (grades I–V). Translational motion and angular variation were measured at each segment from L1–L2 through L5–S1. The relationship between the degree of lumbar disc degeneration and extent of lumbar spine mobility was analyzed. Results. The translational motion in discs with grade I through IV increased gradually, but decreased with grade V. Compared with other less degenerative grades, grade V discs had significantly decreased total intervertebral translational motion (P < 0.05). The angular variation in discs with grade I through IV was fairly constant, but decreased with grade V. Compared with other degenerative grades (I–IV), grade V discs had significantly decreased total intervertebral translational motion (P < 0.05). For less degenerative grades I and II discs, the L2–L3 and L3–L4 segmental units contributed the majority of total angular mobility of the spine. However, for the severely degenerated segments, grade V discs, the contributions of the L2–L3 and L3–L4 significantly decreased (P < 0.01). Conclusion. As disc degeneration developed from the normal to an increasingly severe stage, the motion of lumbar spine progressed from the normal stage to an unstable phase with higher mobility and finally to an ankylosed stage where stability was increased. Level of Evidence: 3

Kinematic relationship between missed ligamentum flavum bulge and degenerative factors in the cervical spine

BACKGROUND CONTEXT Bulging of ligamentum flavum can happen with the aging process and can lead to compression of the spinal cord and nerves. However, the distribution and the risk factors associated with a missed ligamentum flavum bulge (LFB) are unknown. PURPOSE The aim was to evaluate the distribution and risk factors associated with missed LFB in the cervical spine. STUDY DESIGN This was a retrospective analysis of kinematic magnetic resonance images (kMRI). PATIENT SAMPLE Patients diagnosed with symptomatic neck pain or radiculopathy between March 2011 and October 2012 were included. OUTCOME MEASURES The outcome measures were missed LFB and degenerative factors. METHODS A total of 200 patients (1,000 cervical segments) underwent upright kMRI in neutral, flexion, and extension postures. The LFB, sagittal cervical angles, disc herniation, disc degeneration, disc height, angular motion, translational motion, age, and gender were recorded. After excluding segments with LFB in neutral and flexion position, Pearson and Spearman correlation coefficients were used to evaluate the relation between the risk factors and missed LFB in the extension position. RESULTS The average depth of LFB was 0.24±0.71 mm at C2-C3, 1.02±1.42 mm at C3-C4, 1.65±1.48 mm at C4-C5, 2.13±1.37 mm at C5-C6, and 1.05±1.54 mm at C6-C7. The distribution of LFB was the most frequent at C5-C6 level (76.58%) followed by C4-C5 (63.06%). Disc herniation, disc degeneration, angular variation, and translational motion were significantly correlated with missed LFB at C4-C5 andC5-C6. Disc degeneration was the only factor significantly correlated with missed LFB at all cervical segments. CONCLUSIONS Occurrence and depth of missed LFB was the highest at C4-C5 and C5-C6 compared with other cervical levels. Disc degeneration, disc herniation, angular variation, and translational motion could play a role in the development of LFB at C4-C5 andC5-C6.

Secreted phosphoprotein 24 kD inhibits nerve root inflammation induced by bone morphogenetic protein-2.

BACKGROUND CONTEXT Bone morphogenetic protein-2 (BMP-2) has been used to successfully promote spine fusion, but side-effects including nerve inflammation have been observed. PURPOSE To investigate the direct neurotoxic effects of BMP-2 and test the hypotheses that the use of BMP binding proteins, such as secreted phosphoprotein 24 kD (Spp24), can reduce or eliminate these effects. STUDY DESIGN In vitro experiments and in vivo analysis in a rodent model. METHODS In vitro, dorsal root ganglion cells were cultured in the presence of BMP-2 with and without Spp24 and calcitonin gene-related peptide and Substance P, markers of neuroinflammation, were measured by immunohistochemistry. In vivo, rats underwent a left-sided laminotomy at L5 to expose the S1 nerve root and were randomized into four different groups according to the intervention at the laminotomy site: collagen sponge only (no BMP-2 or Spp24), BMP-2 in a collagen sponge only, BMP-2 in a collagen sponge+an empty collagen sponge to act as a barrier, and BMP-2 in a collagen sponge+Spp24 in a collagen sponge to act as a barrier. Functional evaluation was done using the Basso, Beattie, and Bresnahan scale and immunohistochemical analyses were performed using calcitonin gene-related peptide and Substance P staining. RESULTS The neuroinflammatory effects of BMP-2 in vitro were ameliorated by the addition of Spp24. Similarly, in vivo, Spp24 reduced the expression of markers on neuroinflammation in animals treated with BMP-2 and also improved the function after BMP-2 administration. CONCLUSIONS These results confirm that BMP binding proteins have great potential as adjuvant therapies to limit BMP-2 related side-effects in spine surgery.

MicroRNA-300 Regulates the Ubiquitination of PTEN through the CRL4BDCAF13 E3 Ligase in Osteosarcoma Cells

Cullins, critical members of the cullin-RING ubiquitin ligases (CRLs), are often aberrantly expressed in different cancers. However, the underlying mechanisms regarding aberrant expression of these cullins and the specific substrates of CRLs in different cancers are mostly unknown. Here, we demonstrate that overexpressed CUL4B in human osteosarcoma cells forms an E3 complex with DNA damage binding protein 1 (DDB1) and DDB1- and CUL4-associated factor 13 (DCAF13). In vitro and in vivo analyses indicated that the CRL4BDCAF13 E3 ligase specifically recognized the tumor suppressor PTEN (phosphatase and tensin homolog deleted on chromosome 10) for degradation, and disruption of this E3 ligase resulted in PTEN accumulation. Further analyses indicated that miR-300 directly targeted the 3′ UTR of CUL4B, and DNA hypermethylation of a CpG island in the miR-300 promoter region contributed to the downregulation of miR-300. Interestingly, ectopic expression of miR-300 or treatment with 5-AZA-2′-deoxycytidine, a DNA methylation inhibitor, decreased the stability of CRL4BDCAF13 E3 ligase and reduced PTEN ubiquitination. By applying in vitro screening to identify small molecules that specifically inhibit CUL4B-DDB1 interaction, we found that TSC01131 could greatly inhibit osteosarcoma cell growth and could disrupt the stability of the CRL4BDCAF13 E3 ligase. Collectively, our findings shed new light on the molecular mechanism of CUL4B function and might also provide a new avenue for osteosarcoma therapy.

Activation of TNF‐α/NF‐κB axis enhances CRL4BDCAF11 E3 ligase activity and regulates cell cycle progression in human osteosarcoma cells

Cullin 4B, a member of the Cullins, which serve as scaffolds to facilitate the assembly of E3 ligase complexes, is aberrantly expressed in many cancers, including osteosarcoma. Recently, we observed that CUL4B forms the CRL4BDCAF11 E3 ligase, which specifically ubiquitinates and degrades the cyclin‐dependent kinase (CDK) inhibitor p21Cip1 in human osteosarcoma cells. However, the underlying mechanisms regarding the aberrant expression of CUL4B and the upstream members of this signaling pathway are mostly unknown. In this study, we demonstrate that nuclear factor kappaB (NF‐κB) is a direct modulator of CUL4B expression. The CUL4B promoter is responsive to several NF‐κB subunits, including RelA, RelB, and c‐Rel, but not to p50 or p52. Additional studies reveal that the tumor necrosis factor alpha (TNF‐α)/NF‐κB axis pathway is activated in human osteosarcoma cells. This activation causes both CUL4B and NF‐κB subunits to become abundant in the nucleus of human osteosarcoma cells. The down‐regulation of individual genes, including TNFR1, RelA, RelB, c‐Rel, and CUL4B, or pairs of them, including TNFR1 + RelA, TNFR1 + RelB, TNFR1 + c‐Rel, and RelA+CUL4B, has similar effects on cell growth inhibition, colony formation, cell invasion, and in vivo tumor formation, whereas the overexpression of CUL4B in these knockdown cells significantly reverses their phenotypes. The inhibition of the TNF‐α/NF‐κB pathway greatly attenuates CRL4BDCAF11 E3 ligase activity and causes the accumulation of p21Cip1, thereby leading to cell cycle arrest at the S phase. Taken together, our results support a model in which the activation of the TNF‐α/NF‐κB axis contributes to an increase in CRL4BDCAF11 activity and a decrease in p21Cip1 protein levels, thereby controlling cell cycle progression in human osteosarcoma cells.

CRL4B DCAF11 E3 ligase targets p21 for degradation to control cell cycle progression in human osteosarcoma cells

Cell cycle progression in mammals is strictly controlled by a number of cyclin-dependent kinases (CDKs) and CDK inhibitors (CKIs), the expression of which is often dysregulated in cancer cells. Our previous work revealed that Cullin 4B (CUL4B), a critical component of the Cullin4B-RING E3 ligase complex (CRL4B), is overexpressed in human osteosarcoma cells through an unknown mechanism. Here, we demonstrated that CUL4B forms an E3 ligase with RBX1 (RING-box 1), DDB1 (DNA damage binding protein 1), and DCAF11 (DDB1 and CUL4 associated factor 11) in human osteosarcoma cells. In vitro and in vivo ubiquitination analyses indicated that CRL4BDCAF11 E3 ligase was able to specifically ubiquitinate a CDK inhibitor—p21Cip1 at K16, K154, K161 and K163 but not at K75 and K141. Knocking down any component of the CRL4BDCAF11 complex, including CUL4B, DDB1 or DCAF11, using short hairpin RNAs (shRNAs) attenuated the ubiquitination level of p21Cip1, inhibited osteosarcoma cell proliferation, led to cell cycle arrest at S phase, and decreased colony formation rate. Taken together, our data suggest that the CRL4BDCAF11 complex represents a unique E3 ligase that promotes the ubiquitination of p21Cip1 and regulates cell cycle progression in human osteosarcoma cells.

Trends Analysis of rhBMP Utilization in Single-Level Posterior Lumbar Interbody Fusion in the United States

Study Design: Retrospective study. Objectives: Recombinant human bone morphogenetic protein-2 (rhBMP-2) has been widely used in spinal fusion surgery, but there is little information on rhBMP-2 utilization in single-level posterior lumbar interbody fusion (PLIF). The purpose of our study was to evaluate the trends and demographics of rhBMP-2 utilization in single-level PLIF. Methods: Patients who underwent single-level PLIF from 2005 to 2011 were identified by searching ICD-9 diagnosis and procedure codes in the PearlDiver Patient Records Database, a national database of orthopedic insurance records. The year of procedure, age, gender, and region of the United States were recorded for each patient. Results were reported for each variable as the incidence of procedures identified per 100 000 patients searched in the database. Results: A total of 2735 patients had single-level PLIF. The average rate of single-level PLIF with rhBMP-2 maintained at a relatively stable level (28% to 31%) from 2005 to 2009, but decreased in 2010 (9.9%) and 2011 (11.8%). The overall incidence of single-level PLIF without rhBMP-2 (0.68 cases per 100 000 patients) was statistically higher (P < .01) compared to single-level PLIF with rhBMP-2 (0.21 cases per 100 000 patients). The average rate of single-level PLIF with rhBMP-2 utilization was the highest in West (30.1%), followed by Midwest (26.9%), South (20.5%), and Northeast (17.8%). The highest incidence of single-level PLIF with rhBMP-2 was observed in the age group <65 years (0.3 per 100 000 patients). Conclusions: To our knowledge, this is the first study to report on the demographics associated with rhBMP-2 use in single-level PLIF. There was a 3-fold increase in the rate of PLIF without rhBMP-2 compared to PLIF with rhBMP-2, with both procedures being mainly done in patients less than 65 years of age.

Secreted phosphoprotein 24 kD (Spp24) inhibits growth of hepatocellular carcinoma in vivo

Several studies have shown that secreted phosphoprotein 24kD (Spp24) inhibits tumor growth. However, the effects of spp24 on hepatocellular carcinoma are not quite clear. In this study, we observed the inhibitory effect of spp24 on hepatocellular carcinoma in vivo. A subcutaneous hepatocellular carcinoma mice model was established by using Hep G2 cells. After sacrifice at day 40, tumor growth was assessed and tumor cell apoptosis and tumor cells proliferation were assessed by TUNEL assay and immunochemical analysis, respectively. BMP2 slightly stimulated the subcutaneous tumor growth compared with the control. Spp24 significantly inhibited the tumor growth and also abolished the BMP2-induced tumor growth (p<0.05). TUNEL assay and immunochemical analysis further showed that spp24 could enhance tumor cell apoptosis and inhibit cell proliferation (p<0.01). Our data show that spp24 can inhibit the growth of hepatocellular carcinoma. Spp24 may have great potential for cancer treatment.

Distribution of Schmorl nodes in the lumbar spine and their relationship with lumbar disk degeneration and range of motion.

Study Design. A kinematic magnetic resonance imaging study. Objective. To investigate the distribution of Schmorl nodes (SNs) in the lumbar spine in healthy adults, and determine the association with lumbar disk degeneration and lumbar spine motion. Summary of Background Data. SNs have been associated with several pathologies of the lumbar spine, although it has been demonstrated that they also occur in the healthy adult population without a clearly identified cause. A thorough understanding of SN distribution may help reveal reasons for their formation. How disk degeneration and lumbar spine motion relate to SNs is poorly understood. Methods. Kinematic magnetic resonance images (0.6 T) were available for 1179 healthy individuals from 15 to 85 years of age. Spine specialists performed computer-based measurements. All parameters were measured and calculated automatically using the eRAD PACS Viewer (eRAD Inc., version 6.2.1.1). Lumbar disk degeneration was documented according to the Pfirrmann classification system. Lumbar spine lordosis was quantified as the angle between the inferior endplate of L1 and superior endplate of S1. The level of significance was defined as P ⩽ 0.05. The distribution of SNs along the lumbar spine and their relationship with age and sex was investigated using the single factor analysis of variance &khgr;2 test. The relationship between SNs, age group, disk location, and overall grades of lumbar disk degeneration were investigated by multiple logistic regression analysis. Lumbar spine motion was compared between patients with and without SNs via independent t test among 585 individuals with qualified kinematic images. Multiple logistic regression analysis was performed on associations of lumbar motion range among the SN population. Results. The prevalence of SNs in our study population was 28.4%, and SNs were observed to be present more frequently in males (34.6%) than in females (20.2%) (&rgr;< 0.01). There was no significant difference in the incidence of SNs between age groups (&rgr;= 0.18). SNs were more common at the L2 and L3 vertebral bodies (14.3% and 14.4%), whereas SNs were least common at S1 vertebral bodies (1.5%). The highest incidences of SNs presentation was on disks with degeneration grade III (41.9%) and grade IV (45.3%). SN occurrence, aging, and disk location were positively correlated with lumbar disk degeneration grade. The lumbar spine range of motion was significantly different between individuals with and without SNs (31.4°vs. 37.9°, &rgr;< 0.01). The frequency of SNs was associated with decreased lumbar range of motion in all age groups except 51 to 60 years and 61 to 70 years. Conclusion. SNs have a high incidence in individuals without persistent lumbar disorders and were found in disks at all degrees of degeneration. SNs occurrence were positively associated with lumbar disk degeneration In addition, the presence of SNs was correlated with decreased overall lumbar motion across all age groups. Level of Evidence: 1

Effect of Asymmetric Tension on Biomechanics and Metabolism of Vertebral Epiphyseal Plate in a Rodent Model of Scoliosis

To investigate the effect of asymmetric tension on idiopathic scoliosis (IS) and to understand its pathogenic mechanism.