Biomechanical analysis of sandwich vertebrae in osteoporotic patients: finite element analysis.

Objective: The aim of this study was to investigate the biomechanical stress of sandwich vertebrae (SVs) and common adjacent vertebrae in different degrees of spinal mobility in daily life. Materials and methods: A finite element model of the spinal segment of T10-L2 was developed and validated. Simultaneously, T11 and L1 fractures were simulated, and a 6-ml bone cement was constructed in their center. Under the condition of applying a 500-N axial load to the upper surface of T10 and immobilizing the lower surface of L2, moments were applied to the upper surface of T10, T11, T12, L1, and L2 and divided into five groups: M-T10, M-T11, M-T12, M-L1, and M-L2. The maximum von Mises stress of T10, T12, and L2 in different groups was calculated and analyzed. Results: The maximum von Mises stress of T10 in the M-T10 group was 30.68 MPa, 36.13 MPa, 34.27 MPa, 33.43 MPa, 26.86 MPa, and 27.70 MPa greater than the maximum stress value of T10 in the other groups in six directions of load flexion, extension, left and right lateral bending, and left and right rotation, respectively. The T12 stress value in the M-T12 group was 29.62 MPa, 32.63 MPa, 30.03 MPa, 31.25 MPa, 26.38 MPa, and 26.25 MPa greater than the T12 stress value in the other groups in six directions. The maximum stress of L2 in M-T12 in the M-L2 group was 25.48 MPa, 36.38 MPa, 31.99 MPa, 31.07 MPa, 30.36 MPa, and 32.07 MPa, which was greater than the stress value of L2 in the other groups. When the load is on which vertebral body, it is subjected to the greatest stress. Conclusion: We found that SVs did not always experience the highest stress. The most stressed vertebrae vary with the degree of curvature of the spine. Patients should be encouraged to avoid the same spinal curvature posture for a long time in life and work or to wear a spinal brace for protection after surgery, which can avoid long-term overload on a specific spine and disrupt its blood supply, resulting in more severe loss of spinal quality and increasing the possibility of fractures.

Selective versus multi-segmental decompression and fusion for multi-segment lumbar spinal stenosis with single-segment degenerative spondylolisthesis.

BACKGROUND: Lumbar spinal stenosis, often accompanied by degenerative spondylolisthesis, is one of the most common conditions in the elderly. Decompression and fusion is a well-accepted treatment for single-segment lumbar spinal stenosis with degenerative spondylolisthesis; however, the treatment for multi-segment lumbar spinal stenosis with single-segment degenerative spondylolisthesis (MLSS) remains controversial. The objective of this study is to compare the effectiveness of selective decompression and fusion to multi-segmental decompression and fusion for MLSS. METHODS: A total of 42 patients suffering from MLSS who underwent surgery between June 2012 and January 2015 were included in this analysis. Of the 42 patients with minimum 3-year follow-up, 22 underwent selective decompression and fusion, and 20 patients underwent multi-segmental decompression and fusion. Age, gender, symptom duration, operative time, blood loss, the number of decompressed segment and fused segment, and complication were compared between the two groups. The visual analog scale (VAS), Oswestry Disability Index (ODI) and Short Form 36 (SF-36) were used to assess efficacy. RESULTS: Operative time, blood loss, and the number of fused segment in multi-segmental decompression and fusion group were greater than those in selective decompression and fusion group (P < 0.01). The VAS, ODI, and SF-36 scores at 1-year follow-up and 3-year follow-up were significantly improved compared with those preoperatively in both groups (P < 0.01) but were not significantly different between the two groups at each time point (P > 0.05). There was no iatrogenic spinal instability in the decompressed segments in selective decompression and fusion group, while three patients developed postoperative instability at the adjacent segments above the fused segments in multi-segmental decompression and fusion group at 3-year follow-up. CONCLUSIONS: Selective decompression and fusion is a safe and effective method for the treatment of MLSS, with the advantages of shorter operative time, less blood loss, and more preservation of spinal motion segments when compared with multi-segmental decompression and fusion.

miR-148a inhibits pro-inflammatory cytokines released by intervertebral disc cells by regulating the p38/MAPK pathway.

The present study aimed to verify the expression and investigate the role of microRNA (miR)-148a in intervertebral disc degeneration (IDD) and explore the associated underlying mechanisms. Reverse transcription-quantitative polymerase chain reaction (RT-qPCR) was performed to investigate levels of miR-148a in the peripheral blood mononuclear cells (PBMCs) of patients with IDD. To investigate the role of miR-148a in IDD, a stable miR-148a-overexpression/underexpression human nucleus pulposus (NP) cell line was generated by transfection with miR-148a mimic/inhibitor. Then, NP cells were treated with LPS (10 µM) to induce inflammation. The mRNA expression level of miR-148a in NP cells was determined by RT-qPCR and the expression levels of p38 and p-p38 were measured using western blotting. The mRNA expression and supernatant level of pro-inflammatory cytokines, tumor necrosis factor (TNF-α), interleukin (IL)-1ß and IL-6, was evaluated by RT-qPCR and ELISA, respectively. The results indicated that miR-148a was significantly downregulated in the PBMCs of IDD patients compared with healthy controls. In vitro upregulation of miR-148a in LPS-stimulated NP cells, by transfection with miR-148a mimic, resulted in inhibition of p-p38 expression; however, inhibition of miR-148a led to overexpression of p-p38. Meanwhile, the production of pro-inflammatory cytokines (TNF-α, IL-1ß and IL-6) was significantly reduced in miR-148a-overexpressing LPS-stimulated NP cells and significantly increased in miR-148a-underexpressing NP cells. In conclusion, miR-148a inhibits pro-inflammatory cytokines released by intervertebral disc cells via regulation of the p38/mitogen-activated protein kinase pathway.

miR-125b acts as a tumor suppressor in chondrosarcoma cells by the sensitization to doxorubicin through direct targeting the ErbB2-regulated glucose metabolism.

Chondrosarcoma is the second most common type of primary bone malignancy in the United States after osteosarcoma. Surgical resections of these tumors are the only effective treatment to chondrosarcoma patients due to their resistance to conventional chemo- and radiotherapy. In this study, miR-125b was found to perform its tumor-suppressor function to inhibit glucose metabolism via the direct targeting of oncogene, ErbB2. We report miR-125b was downregulated in both chondrosarcoma patient samples and cell lines. The total 20 Asian chondrosarcoma patients showed significantly downregulated miR-125b expression compared with normal tissues. Meanwhile, miR-125 was downregulated in chondrosarcoma cells and doxorubicin resistant cells. Overexpression of miR-125 enhanced the sensitivity of both parental and doxorubicin resistant cells to doxorubicin through direct targeting on the ErbB2-mediated upregulation of glycolysis in chondrosarcoma cells. Moreover, restoration of the expression of ErbB2 and glucose metabolic enzymes in miR-125 pretransfected cells recovered the susceptibility to doxorubicin. Our study will provide a novel aspect on the overcoming chemoresistance in human chondrosarcoma cells and may help in the development of therapeutic strategies for the treatments of patients.

Type-2 cannabinoid receptor regulates proliferation, apoptosis, differentiation, and OPG/RANKL ratio of MC3T3-E1 cells exposed to Titanium particles.

The type-2 cannabinoid receptor (CB2) is expressed in osteoblasts and plays a role in bone metabolism through regulation on bone mass and bone turnover, but the functional importance of CB2 in osteoblasts under Titanium (Ti) stimulation is incompletely understood. This study aimed to investigate the CB2 expression in osteoblasts under Ti stimulation and the effects of CB2 activation on proliferation, apoptosis, differentiation, mineralization, OPG, and RANKL expression of MC3T3-E1 cells exposed to Ti particles. MC3T3-E1 cells were incubated in the presence of Ti particles with or without CB2-specific agonist HU-308 and antagonist SR144528. Ti particles treatment obviously induced the CB2 expression in MC3T3-E1 cells, and reduced the cell survival in a dose- and time-dependent manner (p < 0.05). Addition of HU-308 could dose-dependently alleviate the Ti-induced decrease of cell survival (p < 0.05). The flow cytometry assay showed that comparing with the control group, the apoptosis rate and caspase-3 activity in the Ti group were significantly elevated (p < 0.05), which could be alleviated by HU-308. Moreover, HU-308 effectively attenuated the decrease of cell mineralization capability, alkaline phosphates (ALP) and osteocalcin activity, and increase of OPG/RANKL ratio induced by Ti particles treatment (p < 0.05). These effects were partially counteracted by combined treatment of CB2 antagonist SR144528 (p < 0.05). In conclusion, CB2 activation has a favorable inhibitory effect on Ti-induced reactions in MC3T3-E1 cell through modulating proliferation, apoptosis, differentiation, and RANKL expression. These findings suggest that activation of CB2 might be an effective therapeutic strategy to promote bone formation and reduce bone dissolution.

BDNF-mediated modulation of glycine transmission on rat spinal motoneurons.

BDNF has a widespread distribution in the central and peripheral nervous systems, suggesting that BDNF may play a role in the regulation of motor control. However, the direct actions of BDNF on the motoneurons and their underlying mechanisms are still largely unknown to date. Therefore, by using whole-cell patch clamp recordings, quantitative RT-PCR and immunocytochemistry, the present study was designed to investigate the effects of BDNF on electrical activity and glycinergic transmission on the motoneurons and the underlying receptor mechanism. The results reveal: (i) BDNF did not produce a direct excitatory or inhibitory effect on the motoneurons; (ii) BDNF dose-dependently increased the glycinergic transmission on the motoneurons; (iii) glycinergic transmission on motoneurons was a direct postsynaptic effect; (iv) BDNF-induced enhancement of the glycinergic transmission was mediated by the activation of TrkB receptors; and (v) BDNF and its receptors TrkB had an extensive expression in the motoneurons. These results suggest that BDNF is directly involved in the regulation of glycinergic transmission on the motoneurons through postsynaptic TrkB receptors. Considering that the glycinergic synaptic transmission of motoneurons mainly comes from Renshaw cells, the important inhibitory interneurons of spinal cord, we speculate that BDNF may play an important role in the information integration in the spinal cord and participate in the sensitivity of motoneurons.

Repair of radiation damage of U2OS osteosarcoma cells is related to DNA-dependent protein kinase catalytic subunit (DNA-PKcs) activity.

The present study was to investigate the effects of DNA-PKcs deficiencies on radiation sensitivity of human osteosarcoma U2OS cells to γ-ray and to explore the underlying molecular mechanism. In vitro, U2OS cells were transfected with different DNA-PKcs siRNAs or control siRNAs to establish stably siRNA-transfected cell lines U2OS-Si and U2OS-Sc, respectively. Cell viability and apoptosis after irradiation were analyzed using cell counting kit (CCK-8) and flow cytometric assay, respectively. Expressions of apoptosis-related and oxidative stress-responded proteins were assessed using Western blot. The tumorigenesis activity was examined in nude mice xenograft osteosarcoma mode. Results showed that DNA-PKcs siRNA significantly could inhibit U2OS viability and cell proliferation after exposure to irradiation. Compared with the U2OS and U2OS-Sc cells, the U2OS-Si cells induced higher apoptosis rate and loss of mitochondrial membrane potentials, accompanying with more reactive oxygen species (ROS) and malondialdehyde (MDA) production, increased DNA double-strand breaks (DSBs) induced by irradiation. Protein levels of the anti-apoptotic Bcl-2 were downregulated most obviously in U2OS-Si cells after irradiation, while pro-apoptotic factor Bax and caspase-3 upregulated. Moreover, the antioxidants protein expression levels of Nuclear factor-erythroid 2-related factor 2 (Nrf2) and its target heme oxygenase-1 (HO-1) were also significantly reduced in parallel to DNA-PKcs inhibition in U2OS-Si cells. In nude mice xenograft model, DNA-PKcs siRNA remarkably inhibited tumor growth and dissemination. In conclusion, DNA-PKcs siRNA might have a potential for osteosarcoma treatment by sensitizing osteosarcoma cells to γ-ray through modulation on oxidative stress-mediated DNA DSBs repair and mitochondrial pathway apoptosis.

MiR-27a regulates apoptosis in nucleus pulposus cells by targeting PI3K.

The precise role of apoptosis in the pathogenesis of intervertebral disc degeneration (IDD) remains to be elucidated. We analyzed degenerative nucleus pulposus (NP) cells and found that the expression of miR-27a was increased. The overexpression of miR-27a was further verified using real-time RT-PCR. Bioinformatics target prediction identified phosphoinositide-3 kinases (PI3K) as putative targets of miR-27a. Furthermore, miR-27a inhibited PI3K expression by directly targeting their 3'-UTRs, and this inhibition was abolished by mutation of the miR-27a binding sites. Various cellular processes including cell growth, proliferation, migration and adhesion are regulated by activation of the PI3K/AKT signaling pathway, and nucleus pulposus cells are known to strongly express the phosphorylated survival protein AKT. Our results identify PI3K as a novel target of miR-27a. Upregulation of miR-27a thus targets PI3K, initiating apoptosis of nucleus pulposus cells. This present study revealed that downregulated miR-27a might develop a novel intervention for IDD treatment through the prevention of apoptosis in Nucleus pulposus Cells.

Obesity and risk of hip fracture in adults: a meta-analysis of prospective cohort studies.

BACKGROUND: Many observational studies assessed the association between obesity and risk of hip fracture in adults, but reported controversial results. Our goal was to evaluate the association between obesity and risk of hip fracture in adults by conducting a meta-analysis of prospective cohort studies. METHODS: Three databases, PubMed, Embase and Web of Science, were searched through May 2012 to identify eligible cohort studies. Either a fixed- or a random-effects model was used to calculate the pooled relative risk (RR) with its 95% confidence interval (95%CI). RESULTS: Fifteen prospective cohort studies involving a total 3,126,313 participants were finally included into this meta-analysis. Overall, adults with obesity compared with the normal weight group had a significantly decreased risk of hip fracture (RR: 0.66, 95% CI 0.57 to 0.77, P<0.001). Meta-analyses by the adjusted status of RRs also suggested adults with obesity compared with the reference group had a significantly decreased risk of hip fracture (adjusted RR: 0.48, 95% CI 0.39 to 0.58, P<0.001; unadjusted RR: 0.66, 95% CI 0.56 to 0.78, P<0.001). Subgroup analyses by gender suggested individuals with obesity had a significantly decreased risk for developing hip fracture compared with the reference group in both men (RR 0.54, 95% CI 0.48 to 0.60, P<0.001) and women (RR 0.70, 95% CI 0.58 to 0.84, P<0.001). No evidence of publication bias was observed in this meta-analysis. CONCLUSIONS: This meta-analysis of prospective cohort studies suggests that obesity significantly decreases the risk of hip fracture in adults, and obesity is probably a protective factor of hip fracture in adults.