Scutellarin-mediated autophagy activates exosome release of rat nucleus pulposus cells by positively regulating Rab8a via the PI3K/PTEN/Akt pathway.

To provide a basis for promising exosome-based therapies against intervertebral disc degeneration (IDD), our present research aimed to identify a mechanism underlying the vesicle release from nucleus pulposus cells (NPCs). Scutellarin (SC) is a natural chemotherapeutic agent isolated from Erigeron breviscapus with a variety of biological activities. Here, we observed the significantly elevated autophagy levels in rat NPCs under the stimulation of SC, leading to a concomitant enhancement of intracellular vesicle release, which could be attributed to the inactivation of the phosphoinositide 3-kinase (PI3K)/phosphatase and tensin homolog (PTEN)/protein kinase B (Akt) pathway. To ensure that exosome release was driven by SC via the autophagic pathway, we implemented gain-of-function and loss-of-function studies by additionally using insulin-like growth factor-1 (IGF-1) and small-interfering RNA of autophagy-related gene 5 (ATG5), and the exosome secretion decreased in the case of attenuated autophagy. Evidently, the treatment with SC exerted the remarkable upregulation of Rab8a through the overexpression of ATG5. After the respective knockdown of ATG5 and Rab8a, the increased release of exosomes induced by SC was reversed, whereas the number of intracellular vesicles was restored. Overall, it can be concluded that SC contributes to the autophagy activation in NPCs by acting on the PI3K/PTEN/Akt pathway, which upregulates the expression of Rab8a and promotes the release of exosomes, inspiring novel therapeutic strategies in preventing IDD that might be fruitfully investigated.

Recurrent adamantinoma in the thoracolumbar spine successfully treated by three-level total en bloc spondylectomy by a single posterior approach.

PURPOSE: Adamantinoma is a low-grade primary malignant bone tumour with slow growth and local recurrence. Its occurrence in the spine is extremely rare, particularly with multilevel involvement. This paper wants to present the first case involving a patient with recurrent thoracolumbar spinal adamantinoma, who underwent a successful three-level spondylectomy for en bloc resection. METHODS: A 24-year-old man with osteolytic masses of T11 and T12 vertebral bodies was performed curettage by a posterior approach in 2008. The pathology report showed the excised neoplasm was a rare adamantinoma. This patient underwent a tumorectomy again because of its local recurrence nearly 3 years later. In 2012, it was unfortunately revealed that the excised tumour had relapsed and had spread to the L1 vertebral body. Due to its repeated recurrence and aggressive lesion, total en bloc spondylectomy (TES) for this malignant tumour was thought to be the best option for preventing repeated recurrence and possible cure. TES for T11-L1 thoracolumbar spine was performed and spinal reconstruction was completed with instrumentation and a titanium mesh cage through a one-stage single posterior approach. RESULTS: After three-level TES, neurological deficits of the patient demonstrated good recovery and no evidence of adamantinoma recurrence or deformity was found at 2-year follow-up. CONCLUSIONS: This is the first case involving multilevel thoracolumbar spinal adamantinoma with repeated recurrence to be successfully treated by three-level TES by a single posterior approach.

Molecular actions of ovarian cancer G protein-coupled receptor 1 caused by extracellular acidification in bone.

Extracellular acidification occurs under physiologic and pathologic conditions, such as exercise, ischemia, and inflammation. It has been shown that acidosis has various adverse effects on bone. In recent years there has been increasing evidence which indicates that ovarian cancer G protein-coupled receptor 1 (OGR1) is a pH-sensing receptor and mediates a variety of extracellular acidification-induced actions on bone cells and other cell types. Recent studies have shown that OGR1 is involved in the regulation of osteoclast differentiation, survival, and function, as well as osteoblast differentiation and bone formation. Moreover, OGR1 also regulates acid-induced apoptosis of endplate chondrocytes in intervertebral discs. These observations demonstrate the importance of OGR1 in skeletal development and metabolism. Here, we provide an overview of OGR1 regulation ofosteoclasts, osteoblasts, and chondrocytes, and the molecular actions of OGR1 induced by extracellular acidification in the maintenance of bone health.

Skip-level anterior cervical discectomy and fusion with self-locking stand-alone PEEK cages for the treatment of 2 noncontiguous levels of cervical spondylosis.

STUDY DESIGN: Retrospectively study. OBJECTIVES: To evaluate clinical and radiologic outcomes of skip-level anterior cervical discectomy and fusion (ACDF) with self-locking stand-alone polyetheretherketone (PEEK) cages for the treatment of 2 noncontiguous levels of cervical disk degenerative disease (CDDD). SUMMARY OF BACKGROUND DATA: The use of stand-alone PEEK cages in ACDF has been proved to be safe and effective to treat CDDD. For 2 noncontiguous levels of CDDD, skip-level ACDF with self-locking stand-alone PEEK cages, which fuses only the involved levels without anterior plates, may be the optimal treatment choice. METHODS: Sixteen consecutive patients with 2 noncontiguous levels of CDDD underwent skip-level ACDF with self-locking stand-alone PEEK cages. Clinical outcomes were evaluated using Japanese Orthopaedic Association scores and Odom criteria. Fusion rate and time, cages subsidence, spinal curvature, intervertebral height at the operated level, and adjacent disk degeneration were assessed. RESULTS: Patients were followed up for average 43.6 months (range, 24-78 mo). The mean operative time was 113 minutes (range, 98-134 min) with an average blood loss of 62 mL (range, 47-76 mL). The Japanese Orthopaedic Association score, degree of spinal curvature, and intervertebral height were significantly increased at the final follow-up examination compared with preoperatively (P<0.05). Fifteen patients (93.8%) achieved solid fusion in an average time of 5.1 months. Three cages (9.38%) in 2 patients subsided. Radiologic evidence of adjacent segment degeneration was observed in 3 segments (6.25%; 2 infra-adjacent segments and 1 intermediate segment). No case had neurological deterioration postoperatively. No implant failure or migration was observed during follow-up. CONCLUSIONS: Treatment of 2 noncontiguous levels of CDDD with skip-level ACDF with self-locking stand-alone cages achieved good clinical and radiologic outcomes including a high fusion rate, low complication rate, and excellent maintenance of spinal curvature and intervertebral height.

The relation between zoledronic acid infusion and interbody fusion in patients undergoing transforaminal lumbar interbody fusion surgery.

BACKGROUND: Zoledronic acid (ZOL) has been shown to significantly increase bone mineral density and to decrease the incidence of osteoporotic fractures. However, its safety when used after lumbar interbody fusion surgery remains unclear. We sought to determine whether ZOL infusion 3 days after transforaminal lumbar interbody fusion (TLIF) affects the risk of nonunion. METHODS: This was a randomized, double-blind, placebo-controlled trial involving subjects who underwent TLIF surgery. Eighty-two subjects (≥50 years of age) were randomly assigned to receive either 5 mg intravenous ZOL (N=41) or placebo (N=41) 3 days after surgery. Each patient received a lumbar computed tomography scan 6 months and 12 months postoperatively. We evaluated interbody fusion using the multiplanar reconstruction technique. Clinical outcome was evaluated with the Oswestry Disability Index. Bone turnover markers (amino terminal propeptides of type I collagen and C-telopeptide of type I collagen) were measured to investigate the biological effects of ZOL on spinal fusion. RESULTS: In the ZOL group, 7 levels (11.5%) exhibited non-union; in the placebo group, 9 levels (14.5%) exhibited nonunion at 12 months postoperatively. This difference was not statistically significant (P=0.82). The difference in ODI scores between two groups was not statistically significant at any of the follow-up times. However, ZOL decreased bone turnover markers significantly. CONCLUSIONS: There was no association between ZOL treatment and nonunion of the lumbar spinal bone. Thus, undergoing lumbar interbody fusion surgery is not a valid reason to suspend or avoid treatment with ZOL.

[Comparison of percutaneous versus open monosegment instrumentation in the treatment of incomplete thoracolumbar burst fracture].

OBJECTIVE: To compare the safety and efficacies of traditional open versus percutaneous monosegmental pedicle screw fixation in the treatment of incomplete thoracolumbar spinal fracture. METHODS: A retrospective analysis was conducted for 44 inpatients with a diagnosis of incomplete thoracolumbar spinal fracture (AO classification: A3.1 and A3.2) undergoing monosegmental pedicle instrumentation (MSPI) from September 2008 to January 2011. There were 24 cases in percutaneous group and 20 cases in traditional open group. The mean operative duration, blood loss, blood drainage, visual analogue scale/score (VAS) and vertebral kyphotic angle at pre- and post-operation were evaluated. RESULTS: No significant differences existed in operative durations between two groups (P > 0.05). Significant differences between two groups were observed in terms of intra-operative blood loss and VAS scores at Week 1 postoperation (P < 0.05). There were no significant differences in VAS score preoperation, 1 year postoperation or pre-and post-operative vertebral kyphotic angle (P > 0.05). No complications of iatrogenic neurological injury or hardware failure occurred. CONCLUSION: The application of percutaneous monosegment pedicle instrumentation in the treatment of thoracolumbar fractures in type of A 3.1 and A 3.2 is both feasible and safe. Its postoperative therapeutic effect is comparable to that of traditional open monosegmental fixation.

Excessive mechanical strain accelerates intervertebral disc degeneration by disrupting intrinsic circadian rhythm.

Night shift workers with disordered rhythmic mechanical loading are more prone to intervertebral disc degeneration (IDD). Our results showed that circadian rhythm (CR) was dampened in degenerated and aged NP cells. Long-term environmental CR disruption promoted IDD in rats. Excessive mechanical strain disrupted the CR and inhibited the expression of core clock proteins. The inhibitory effect of mechanical loading on the expression of extracellular matrix genes could be reversed by BMAL1 overexpression in NP cells. The Rho/ROCK pathway was demonstrated to mediate the effect of mechanical stimulation on CR. Prolonged mechanical loading for 12 months affected intrinsic CR genes and induced IDD in a model of upright posture in a normal environment. Unexpectedly, mechanical loading further accelerated the IDD in an Light-Dark (LD) cycle-disrupted environment. These results indicated that intrinsic CR disruption might be a mechanism involved in overloading-induced IDD and a potential drug target for night shift workers.

TNF-α-stimulated nucleus pulposus cells induce cell apoptosis through the release of exosomal miR-16 targeting IGF-1 and IGF-1R in rats.

BACKGROUND: Exosomes may contain excess cellular components released by cells in response to harmful external stimuli to maintain cellular homeostasis. Inflammatory cytokines, such as tumor necrosis factor-alpha (TNF-α), can induce cell apoptosis, alter cellular component expression levels, and stimulate exosome release. In this study, we examined whether exosomes released from nucleus pulposus cells (NPCs) under inflammatory conditions could induce normal NP cell apoptosis in rats and its underlining mechanism. METHODS: Exosomes were isolated from TNF-α-treated NPCs and used to treat normal NPCs. The effects were assessed by flow cytometry and western blot analysis. Anti-apoptotic insulin-like growth factor-1 (IGF-1) expression in NPCs was assessed by western blot analysis. Given the exosomal miRNAs might be the key factors of exosomes, bioinformatics approaches and quantitative real-time polymerase chain reaction (qRT-PCR) were used to identify IGF-1-regulating micro RNAs (miRNAs), including miR-16. Luciferase reporter assay assessed miR-16 regulation of IGF-1 and IGF-1 receptor (IGF-1R). NPCs were transfected with miR-16 mimic, and exosomes were applied to normal NPCs. NPCs were pretreated with 10 ng/mL TNF-α, transfected with miR-16 inhibitors, and the exosomes were isolated. Cell and exosome miR-16 levels were detected by qRT-PCR. Western blot analysis determined IGF-1, IGF-1R, and apoptotic marker levels in exosome-treated NPCs. RESULTS: Exosomes from TNF-α-treated NPCs induced apoptosis in normal NPCs and repressed IGF-1 expression. Exosomal miR-16 regulated IGF-1 and induced NPC apoptosis. The dual-luciferase reporter assay revealed that miR-16 binds the 3' untranslated regions (3'-UTRs) of IGF-1 and IGF-1R. Exosomal miR-16 repressed IGF-1 and the IGF-1R/phosphoinositide 3-kinase (PI3K)/protein kinase B (Akt) pathway which therefore induced NPC apoptosis. Rescue experiments using miR-16 inhibitors further validated these findings. CONCLUSIONS: The inflammatory factor TNF-α stimulated exosome release from NPCs, which induced the apoptosis of normal NPCs through the actions of exosomal miR-16. Exosomal miR-16 directly repressed the anti-apoptotic IGF-1/IGF-1R pathway, increasing the apoptosis of NPCs.

Decorin inhibits nucleus pulposus apoptosis by matrix-induced autophagy via the mTOR pathway.

Decorin (Dcn) is a member of the class I small leucine-rich proteoglycans, whose expression in the nucleus pulposus (NP) of intervertebral discs (IVDs) has been shown to increase with aging in humans and sheep. Dcn induces autophagy in endothelial cells; however, its precise role in NP and IVD degeneration during aging is not well understood. We addressed this question in the present study by treating rat nucleus pulposus cells (NPCs) with different concentrations of Dcn. The Western blot analysis and terminal deoxynucleotidyl transferase dUTP nick end labeling assay results showed that Dcn treatment induced autophagy and decreased apoptosis caused by interleukin (IL)-1ß application. This effect was dependent on the protein kinase B/mechanistic target of rapamycin (mTOR)/p70 S6 kinase signaling. Dcn treatment also decreased the expression of matrix metalloproteinase-3 and -13 and decreased the IL-1ß-induced attenuation of collagen type II and aggrecan levels. The role of Dcn in stimulating autophagy was further supported by the fact that the observed effects were abrogated by knocking down autophagy-related protein 7 with Atg7 small interfering RNA. Thus, Dcn protects NPCs in IVDs from IL-1ß-induced apoptosis and degeneration by promoting autophagy through mTOR signaling.

Hsa_circ_0006571 promotes spinal metastasis through sponging microRNA-138 to regulate sirtuin 1 expression in lung adenocarcinoma.

BACKGROUND: Circular RNAs (circRNAs) are known to participate in lung cancer. However, their role in spinal metastasis (SM) of lung adenocarcinoma remains elusive. In this study, we determined that hsa_circ_0006571 serves as a sponge for miR-138, which targets sirtuin 1 (Sirt1) in the development of SM. METHODS: A human circRNA microarray was performed to compare SM and lung adenocarcinoma samples. The expression of hsa_circ_0006571 and miR-138 was determined using quantitative polymerase chain reaction (qPCR) in vitro and in vivo. Cell proliferation was performed by Cell Counting Kit-8 (CCK-8) and apoptosis was analyzed by Annexin V/PI staining. RNA-pulldown and RNA immunoprecipitation (RIP) were used to analyze the interaction between hsa_circ_0006571. Tumor metastasis was determined through a xenograft experiment in vivo. RESULTS: Hsa_circ_0006571 was observed to be significantly upregulated in SM tissues through circRNA microarray and qPCR. We detected a lower expression of miR-138 in SM tissues compared with lung adenocarcinoma. Hsa_circ_0006571 silencing suppressed lung cancer cell proliferation and migration while promoting apoptosis. Hsa_circ_0006571 interacted with miR-138 to promote expression of Sirt1, leading to activation of epithelial-mesenchymal transition (EMT). Xenograft experiments showed that downregulation of hsa_circ_0006571 delayed the SM of lung adenocarcinoma cells via the miR-138-Sirt1 axis. CONCLUSIONS: Hsa_circ_0006571 promoted tumor cell migration and invasion via the miR-138/Sirt1 pathway. Our observations indicate that circRNAs are possible novel therapeutic targets for SM of lung adenocarcinoma.

Degenerative Spondylolisthesis in the Fifth Lumbar Vertebra and Radiographic Parameters: A Correlation Analysis.

STUDY DESIGN: A retrospective study. OBJECTIVE: This study aimed to analyze the relationships between degenerative spondylolisthesis in the fifth lumbar vertebra (L5-DS) and radiographic parameters and to further determine the radiographic predictors of the development of L5-DS. SUMMARY OF BACKGROUND DATA: Degenerative lumbar spondylolisthesis (DLS) is a common degenerative disease of the spine; however, the correlations between L5-DS and radiographic parameters remain controversial. PATIENTS AND METHODS: This retrospective case-control study was conducted in our hospital. Between 2011 and 2014, a total of 84 subjects with L5-DS were enrolled in the DLS group, and 56 healthy volunteers were recruited to the control group. A series of radiographic parameters, including the bone mineral density, disk degenerative index, disk height, L5 vertebral size (L5-VS), lumbar lordosis angle (LL), sacral slope angle (SS), pelvic incidence (PI), facet joint angulations (FJA) of the cephalad and caudad portions, and asymmetry of the FJA, were measured in both groups by 3 examiners. RESULTS: The bone mineral density, disk degenerative index, disk height, L5-VS, LL, SS, PI, and FJA exhibited significant differences (P=0.014-0.045) between the DLS and control groups. Significant changes in the FJA of the cephalad and caudad portions in the L4-L5 and L5-S1 segments were observed between the 2 groups (P=0.00, 0.00), whereas no significant differences in the asymmetries of FJA were observed in the L4-L5 or L5-S1 segments (P=0.605-0.972). Among all of the parameters, the L5-VS (P=0.025), SS (P=0.020), LL (P=0.031), PI (P=0.014), and FJA (P=0.022) were identified as being associated with the DLS group by multiple logistic regression analysis. CONCLUSIONS: In this study, SS, LL, PI, and a more sagittal FJA were proven to be risk factors for L5-DS, whereas L5-VS was found to be a likely protective factor against L5-DS. These parameters should be considered predictors of L5-DS.

Bioabsorbable self-retaining PLA/nano-sized ß-TCP cervical spine interbody fusion cage in goat models: an in vivo study.

STUDY DESIGN: This is an experimental animal study. OBJECTIVE: The objective of this study was to compare an anterior cervical discectomy and interbody fusion of a novel polylactide/nano-sized ß-tricalcium phosphate (PLA/nß-TCP) bioabsorbable self-retaining cervical fusion cage (BCFC) with an autologous bone graft and polyetheretherketone (PEEK) cages. BACKGROUND: Although PLA cervical cages have potential advantages compared with traditional materials, they are not currently routinely used in spine surgery because of undesirable effects such as the lack of osteoconductivity and osteolysis around the implant. This study involved the manufacturing of a bioabsorbable cage from PLA/nß-TCP that was then used as a device for anterior cervical discectomy and fusion (ACDF) on a goat cervical spine fusion model. MATERIALS AND METHODS: Eighteen goats underwent C3/C4 discectomy and were randomly divided into three groups based on the following methods: Group A (n=6), an autologous bone graft; Group B (n=6), PEEK cage filled with an autologous graft; and Group C (n=6), BCFC filled with an autologous iliac bone. Radiography was performed preoperatively and postoperatively and at 1, 4, 8, and 12 weeks after the operation. Disc space height (DSH) was measured at the same time. After 12 weeks, the fused segments were harvested and evaluated with functional radiographic views, biomechanical testing, and histological analyses. RESULTS: Over a 12-week period, the BCFC and PEEK cage groups exhibited significantly higher DSH values than the bone graft group. Additionally, the BCFC group yielded a significantly lower range of motion in axial rotation than both the autologous bone graft and PEEK cage groups. A histologic evaluation revealed an increased intervertebral bone volume/total volume ratio and better interbody fusion in the BCFC group than in the other groups. CONCLUSION: The BCFC device exhibited better results than the autologous bone graft and PEEK cages in single-level ACDF models in vivo. This device may be a potential alternative to the current PEEK cages.

Minimum 5 year follow-up of multi-segmental lumbar degenerative disease treated with discectomy and the Wallis interspinous device.

We evaluate the clinical effects and radiological findings of the Wallis interspinous device (Zimmer, Warsaw, IN, USA) for the treatment of multi-segmental lumbar degenerative disease after a minimum 5 year follow-up period. A total of 26 adult patients underwent a primary discectomy followed by fixation of the segment with the Wallis interspinous device between December 2007 and August 2008. Twelve men and 14 women with an age range of 43 to 56 years (average: 47.6) were included. The visual analogue scale (VAS) for low back and leg pain, Oswestry Disability Index (ODI), foraminal height (FH), anterior disc height (aDH) and posterior disc height (pDH), range of motion (ROM) and Pfirrmann grades were obtained and compared before and after surgery. The VAS and ODI significantly decreased postoperatively (p < 0.05). The postoperative FH and pDH values increased significantly compared with the preoperative levels (p < 0.01) and the increase in the FH and pDH values remained statistically significant during the follow-up period. There were no statistically significant changes in the aDH values before and after surgery (p > 0.05). Also, there were no statistically significant changes in the ROM and Pfirrmann grade at the instrumented level and at the cephalad-adjacent segment (p>0.05). In our study, no patient underwent further surgery because of a re-prolapse or progression of index level degeneration or adjacent segment disease. The Wallis interspinous device was a useful alternative for treating multi-segmental lumbar degenerative disease and it offered a significant minimum 5 year symptom control.

Establishment of intervertebral disc degeneration model induced by ischemic sub-endplate in rat tail.

BACKGROUND CONTEXT: Microcirculatory dysfunction of the sub-endplate is considered to reduce nutrient supply to the intervertebral disc (IVD); however, direct interruption or destruction of blood vessels in the bone marrow of the vertebrae body adjacent to the endplate has not yet been described, especially with regard to the calcification and ossification of the cartilaginous endplate occurring during IVD degeneration. PURPOSE: The purpose of the study was to evaluate the causal relationship between IVD degeneration and blocking of the main blood supply gateway through the endplate. STUDY DESIGN/SETTING: The study describes a new IVD degeneration model induced by ischemic sub-endplate. PATIENT SAMPLE: A total of 40 Sprague-Dawley rats were included in the study group. OUTCOME MEASURES: To assess disc height, a radiograph was taken each month for 4 months. Changes in endplate, nucleus pulposus (NP), and annulus fibrosus (AF) were evaluated by histochemical and immunohistochemical staining to detect IVD degeneration. METHODS: Injection of 30 µL absolute ethanol into the IVD of rat tail at Co7/Co8 was used to induce injury. Controls were injected with 30 µL of phosphate-buffered saline into the IVD at Co8/Co9. RESULTS: In the ethanol-injected group, disc height gradually decreased and bone sclerosis developed in the endplate. In the NP, cell transformation occurred, changing from predominantly vacuolar cells to chondrogenic cells and eventually fibrocartilaginous cells, along with fibrosis of the NP. As degeneration progressed, the AF developed disordered morphology and rough lamellae, and eventually ruptures and fibrosis. The extent of degeneration increased gradually over time, while the wavy tidemark of the growth plate regressed, and eventually disappeared. Initially positive collagen type II staining gradually decreased on the ischemic side of the sub-endplate. Except at the 3-month time point, expression of collagen type II, aggrecan, and Sox-9 in NP decreased gradually as degeneration progressed, compared with the control group. CONCLUSIONS: This model successfully reproduced IVD degeneration, which could be used for etiological studies on IVD degeneration and investigation of nutrient supply disturbance, and may provide a theoretical foundation for clinical intervention and therapy for IVD degeneration in the future.

Polypyrrole coating on poly-(lactide/glycolide)-ß-tricalcium phosphate screws enhances new bone formation in rabbits.

Polypyrrole (PPy) has gained interest as an implant material due to its multifunctional properties and its high compatibility with several cell and tissue types. For the first time, the biocompatibility and osteointegration of PPy coating, incorporated with chondroitin sulfate (CS), were studied in vivo by implanting PPy-coated bioabsorbable bone fixation composite screws of poly-(lactide/glycolide) copolymer (PLGA) and ß-tricalcium phosphate (TCP) into New Zealand white rabbits. Uncoated bioabsorbable polymer composite screws and commercially available stainless steel cortical screws were used as reference implants. The rabbits were euthanized 12 and 26 weeks after the implantation. The systemic effects were evaluated from food and water consumption, body weight, body temperature, clinical signs, blood samples, internal organ weights, and histological examination. Local effects were studied from bone tissue and surrounding soft tissue histology. New bone formation was evaluated by micro-computed tomography, tetracycline labeling and torsion tests. Torsion tests were performed in order to capture the peak value of the torsion force during the course of the screw's loosening. The coated screws induced significantly more bone formation than the uncoated screws. In addition, none of the implants induced any systemic or local toxicity. The results suggest that PPy is biocompatible with bone tissue and is a potential coating for enhancing osteointegration in orthopedic implants.

Ovarian cancer G protein-coupled receptor 1 is involved in acid-induced apoptosis of endplate chondrocytes in intervertebral discs.

Ovarian cancer G protein-coupled receptor 1 (OGR1) has been shown to be a receptor for protons. We investigated the role of proton-sensing G protein-coupled receptors in the apoptosis of endplate chondrocytes induced by extracellular acid. The expression of proton-sensing G protein-coupled receptors was examined in rat lumbar endplate chondrocytes. Knockdown of OGR1 was achieved by transfecting chondrocytes with specific short hairpin RNA (shRNA) for OGR1. Apoptotic changes were evaluated by DNA fragmentation ELISA, electron microscopy, and flow cytometry. Intracellular calcium ([Ca(2+) ]i) was analyzed with laser scanning confocal microscopy. The mechanism of OGR1 in acid-induced apoptosis of endplate chondrocytes was also investigated. We found that OGR1 was predominantly expressed in rat endplate chondrocytes, and its expression was highly upregulated in response to acidosis. Knocking down OGR1 with shRNAs effectively attenuated acid-induced apoptosis of endplate chondrocytes and increased [Ca(2+) ]i. Blocking OGR1-mediated [Ca(2+) ]i elevation inhibited acid-induced calcium-sensitive proteases such as calpain and calcineurin, and also inhibited the activation of Bid, Bad, and Caspase 3 and cleavage of poly (ADP-ribose) polymerase (PARP). OGR1-mediated [Ca(2+) ]i elevation has a crucial role in apoptosis of endplate chondrocytes by regulating activation of calcium-sensitive proteases and their downstream signaling.

Biomechanical stability of a bioabsorbable self-retaining polylactic acid/nano-sized ß-tricalcium phosphate cervical spine interbody fusion device in single-level anterior cervical discectomy and fusion sheep models.

PURPOSE: The aim of this study was to investigate the biomechanical stability provided by a novel, polylactic acid/nano-sized, ß-tricalcium phosphate, bioabsorbable, self-retaining cervical fusion cage (BCFC). METHODS: Quasistatic nonconstraining torques (maximum 1.5 NM) induced flexion, extension, lateral bending (±1.5 NM), and axial rotation (±1.5 NM) on 32 sheep cervical spines (C2-C5). The motion segment C3-C4 was first tested intact; the following groups were tested after complete discectomy: autologous tricortical iliac crest bone graft, Medtronic-Wego polyetheretherketone (PEEK) cage, Solis PEEK cage, and BCFC. The autologous bone graft group was tested with an anterior plate. The mean range of motion (ROM) was calculated from the load-displacement curves. RESULTS: BCFC significantly decreased ROM in lateral bending and axial rotation compared to other implants, and no significant difference in ROM between two types of PEEK cages and BCFC could be observed in flexion and extension. Anterior cervical plate (ACP) significantly decreased ROM in flexion and extension, but no significant difference in ROM between BCFC and bone graft plus ACP could be determined in lateral bending and axial rotation. CONCLUSION: The BCFC device showed better stability to autologous tricortical iliac crest bone graft and PEEK cages in single-level anterior cervical discectomy and fusion models and thus may be a potential alternative to the current PEEK cages.

Degradation and osteogenic potential of a novel poly(lactic acid)/nano-sized ß-tricalcium phosphate scaffold.

The purpose of this study was to investigate the influence of nano-sized ß-tricalcium phosphate (ß-TCP) on the biological performance of poly (lactic acid) (PLA) composite scaffolds by using in vitro degradation and an in vivo model of heterotopic bone formation. Nano-sized ß-TCP (nß-TCP) was prepared with a wet grinding method from micro-sized ß-TCP (mß-TCP), and composite scaffolds containing 0, 10, 30, or 50 wt% nß-TCP or 30 wt% mß-TCP were generated using a freeze-drying method. Degradation was assessed by monitoring changes in microstructure, pH, weight, and compressive strength over a 26-week period of hydrolysis. Composite scaffolds were processed into blocks, and implanted into muscular pockets of rabbits after loading with recombinant human bone morphogenetic protein-2 (rhBMP-2). New bone formation was evaluated based on histological and immunohistochemical analysis 2, 4, and 8 weeks after implantation. The in vitro results indicated that the buffering effect of nß-TCP was stronger than mß-TCP, which was positively correlated with the content of nß-TCP. The in vivo findings demonstrated that nß-TCP enhanced the osteoconductivity of the scaffolds. Although composite scaffolds containing 30% nß-TCP exhibited similar osteoconductivity to 50% nß-TCP, they had better mechanical properties than the 50% nß-TCP scaffolds. This study supports the potential application of a composite scaffold containing 30% nß-TCP as a promising scaffold for bone regeneration.