Manipulation of osteogenic and adipogenic differentiation of human degenerative disc and ligamentum flavum derived progenitor cells using IL-1ß, IL-19, and IL-20.

PURPOSE: To elucidate whether pro-inflammatory cytokines might influence the commitment of intervertebral disc (IVD)- and ligamentum flavum (LF)-derived progenitor cells toward either osteogenesis or adipogenesis, specifically Interleukin-1ß (IL-1ß), IL-19, and IL-20. METHODS: Sixty patients with degenerative spondylolisthesis and lumbar or lumbosacral spinal stenosis were included in the study. Injuries to the spine, infections, and benign or malignant tumors were excluded. From nine patient samples, IVD- and LF-derived cells were isolated after primary culture, and two clinical samples were excluded due to mycoplasma infection. The effects of IL-1ß, IL-19, as well as IL-20 in regulating osteogenic and adipogenic differentiation in vitro were investigated. RESULTS: Primary IVD- and LF-derived cells were found to have a similar cell morphology and profile of surface markers (CD44, CD90, and CD105) as placenta-derived mesenchymal stem cells (MSCs). Primary IVD/LF cells have a high capacity to differentiate into osteocytes and adipocytes. IL-19 had a tendency to promote adipogenesis. IL-20 inhibited osteogenesis and promoted adipogenesis; IL-1ß promoted osteogenesis but inhibited adipogenesis. CONCLUSION: IL-1ß, IL-19, and IL-20 impact the adipogenic and osteogenic differentiation of IVD-derived and LF-derived cells. Modulating the expression of IL-1ß, IL-19, and IL-20 provides a potential avenue for controlling cell differentiation of IVD- and LF-derived cells, which might have beneficial effect for degenerative spondylolisthesis and spinal stenosis.

Biomechanical feasibility of semi-rigid stabilization and semi-rigid lumbar interbody fusion: a finite element study.

BACKGROUND: Semi-rigid lumbar fusion offers a compromise between pedicle screw-based rigid fixation and non-instrumented lumbar fusion. However, the use of semi-rigid interspinous stabilization (SIS) with interspinous spacer and ligamentoplasty and semi-rigid posterior instrumentation (SPI) to assist interbody cage as fusion constructs remained controversial. The purpose of this study is to investigate the biomechanical properties of semi-rigidly stabilized lumbar fusion using SIS or SPI and their effect on adjacent levels using finite element (FE) method. METHOD: Eight FE models were constructed to simulate the lumbosacral spine. In the non-fusion constructs, semi-rigid stabilization with (i) semi-rigid interspinous spacer and artificial ligaments (PD-SIS), and (ii) PI with semi-rigid rods were simulated (PD + SPI). For fusion constructs, the spinal models were implanted with (iii) PEEK cage only (Cage), (iv) PEEK cage and SIS (Cage+SIS), (v) PEEK cage and SPI (Cage+SPI), (vi) PEEK cage and rigid PI (Cage+PI). RESULT: The comparison of flexion-extension range of motion (ROM) in the operated level showed the difference between Cage+SIS, Cage+SPI, and Cage+PI was less than 0.05 degree. In axial rotation, ROM of Cage+SIS were greater than Cage+PI by 0.81 degree. In the infrajacent level, while Cage+PI increased the ROM by 24.1, 27,7, 25.9, and 10.3% and Cage+SPI increased the ROM by 26.1, 30.0, 27.1, and 10.8% in flexion, extension, lateral bending and axial rotation respectively, Cage+SIS only increased the ROM by 3.6, 2.8, and 11.2% in flexion, extension, and lateral bending and reduced the ROM by 1.5% in axial rotation. The comparison of the von Mises stress showed that SIS reduced the adjacent IVD stress by 9.0%. The simulation of the strain energy showed a difference between constructs less than 7.9%, but all constructs increased the strain energy in the infradjacent level. CONCLUSION: FE simulation showed semi-rigid fusion constructs including Cage+SIS and Cage+SPI can provide sufficient stabilization and flexion-extension ROM reduction at the fusion level. In addition, SIS-assisted fusion resulted in less hypermobility and less von Mises stress in the adjacent levels. However, SIS-assisted fusion had a disadvantage of less ROM reduction in lateral bending and axial rotation. Further clinical studies are warranted to investigate the clinical efficacy and safety of semi-rigid fusions.

Mechanical Stretch-Induced NLRP3 Inflammasome Expression on Human Annulus Fibrosus Cells Modulated by Endoplasmic Reticulum Stress.

Excessive mechanical loading is a major cause of spinal degeneration, typically originating from a tear in the annulus fibrosus (AF). Endoplasmic reticulum (ER) stress and NLRP3 (NOD-, LRR- and pyrin domain-containing protein 3) inflammasome have been implicated in the pathogenesis of intervertebral disc (IVD) degeneration. However, the causal relationship between the mechanical stretching of AF cells and the NLRP3 inflammasome response associated with ER stress remains scarce. To elucidate the pathogenesis and regulatory mechanisms of mechanical stretch-induced IVD degeneration, human AF cell lines were subjected to different degrees of cyclic stretching to simulate daily spinal movements. Our results indicated that 15% high cyclic stretch (HCS) induced the expression of NLRP3 and interleukin-1 beta (IL-1ß) and was also responsible for the increased expression of NADPH (nicotinamide adenine dinucleotide phosphate) oxidase 2 (NOX2) and reactive oxygen species (ROS) in human AF cells. In addition, HCS increased the expression of glucose-regulated protein 78 (GRP78), an ER stress chaperone, which was neutralized with tauroursodeoxycholic acid (TUDCA), an ER stress inhibitor. In addition, HCS was found to induce thioredoxin-interacting protein (TXNIP) expression and NLRP3 inflammasome activation, which can be suppressed by si-NOX2 or the NOX2 inhibitor GSK2795039. Consequently, HCS upregulated ER stress and ROS production, leading to increased NLRP3 and IL-1ß expression in human AF cells, and may further accelerate IVD degeneration.

Comparison of Posterior Fixation Strategies for Thoracolumbar Burst Fracture: A Finite Element Study.

The management of thoracolumbar (TL) burst fractures remained challenging. Due to the complex nature of the fractured vertebrae and the lack of clinical and biomechanical evidence, currently, there was still no guideline to select the optimal posterior fixation strategy for TL burst fracture. We utilized a T10-L3 TL finite element model to simulate L1 burst fracture and four surgical constructs with one- or two-level suprajacent and infrajacent instrumentation (U1L1, U1L2, U2L1, and U2L2). This study was aimed to compare the biomechanical properties and find an optimal fixation strategy for TL burst fracture in order to minimize motion in the fractured level without exerting significant burden in the construct. Our result showed that two-level infrajacent fixation (U1L2 and U2L2) resulted in greater global motion reduction ranging from 66.0 to 87.3% compared to 32.0 to 47.3% in one-level infrajacent fixation (U1L1 and U2L1). Flexion produced the largest pathological motion in the fractured level but the differences between the constructs were small, all within 0.26 deg. Comparisons in implant stress showed that U2L1 and U2L2 had an average 25.3 and 24.8% less von Mises stress in the pedicle screws compared to U1L1 and U1L2, respectively. The construct of U2L1 had better preservation of the physiological spinal motion while providing sufficient range of motion reduction at the fractured level. We suggested that U2L1 is a good alternative to the standard long-segment fixation with better preservation of physiological motion and without an increased risk of implant failure.

MLN4924, a Protein Neddylation Inhibitor, Suppresses the Growth of Human Chondrosarcoma through Inhibiting Cell Proliferation and Inducing Endoplasmic Reticulum Stress-Related Apoptosis.

Chondrosarcoma, a heterogeneous malignant bone tumor, commonly produces cartilage matrix, which generally has no response to conventional therapies. Studies have reported that MLN4924, a NEDD8-activating enzyme inhibitor, achieves antitumor effects against numerous malignancies. In this study, the suppressive effects of MLN4924 on human chondrosarcoma cell lines were investigated using in vitro and in vivo assays, which involved measuring cell viability, cytotoxicity, apoptosis, proliferation, cell cycles, molecule-associated cell cycles, apoptosis, endoplasmic reticulum (ER) stress, and tumor growth in a xenograft mouse model. Our results demonstrated that MLN4924 significantly suppressed cell viability, exhibited cytotoxicity, and stimulated apoptosis through the activation of caspase-3 and caspase-7 in chondrosarcoma cell lines. Furthermore, MLN4924 significantly inhibited cell proliferation by diminishing the phosphorylation of histone H3 to cause G2/M cell cycle arrest. In addition, MLN4924 activated ER stress⁻related apoptosis by upregulating the phosphorylation of c-Jun N-terminal kinase (JNK), enhancing the expression of GRP78 and CCAAT-enhancer-binding protein homologous protein (CHOP, an inducer of endoplasmic ER stress⁻related apoptosis) and activating the cleavage of caspase-4. Moreover, MLN4924 considerably inhibited the growth of chondrosarcoma tumors in a xenograft mouse model. Finally, MLN4924-mediated antichondrosarcoma properties can be accompanied by the stimulation of ER stress⁻related apoptosis, implying that targeting neddylation by MLN4924 is a novel therapeutic strategy for treating chondrosarcoma.

Evaluation of the effect of different sitting assistive devices in reclining wheelchair on interface pressure.

BACKGROUND: Reclining wheelchair users often add one or more sitting assistive devices to their wheelchairs, but the effect of these additional sitting assistive devices on the risk of pressure ulcers has rarely been investigated. This study examined the four modes of reclining wheelchair without and with different sitting assistive devices, namely the back reclined mode, the lumbar support with back reclined mode, the femur upward with back reclined mode, and the lumbar support with femur upward with back reclined mode, in terms of their effects on human-wheelchair interface pressure. METHODS: This study recruited 16 healthy participants to undergo the aforementioned four modes in random order and have their human-wheelchair interface pressure measured. The initial setting of experimental reclining wheelchair backrest was pushed backward to reach a 150° recline. The data on interface pressure were collected for 5 s while the participant maintained a stable sitting position. The contact area, average pressure, and peak pressure on the back area, ischial area, and femur area were recorded and calculated. RESULTS: Among all tested modes, the lumbar support with femur upward with back reclined mode provided the most significant reduction in stress load on the ischial area (P ≤ 0.010) and shifted part of the load to the femur area (P ≤ 0.009). CONCLUSIONS: This study quantified the effects of and differences between various reclining wheelchair-sitting assistive device combination modes. These findings are useful for the decision-making processes of rehabilitation physicians, wheelchair users, and manufacturers.

Reactive oxygen species mediate Terbufos-induced apoptosis in mouse testicular cell lines via the modulation of cell cycle and pro-apoptotic proteins.

Terbufos (S-t-butylthiomethyl-O,O-diethyl phosphorodithioate) is a highly toxic organophosphate which is extensively used as an insecticide and nematicide. Chronic exposure to terbufos causes neuronal injury and predisposes to neurodegenerative diseases. Accumulating evidence has shown that the exposure to terbufos, as an occupational risk factor, may also cause reproductive disorders. However, the exact mechanisms of reproductive toxicity remain unclear. The present study aimed to investigate the toxic effect of terbufos on testicular cells and to explore the mechanism of toxicity on a cellular level. The cytotoxic effects of terbufos on mouse immortalized spermatogonia (GC-1), spermatocytes (GC-2), Leydig (TM3), and Sertoli (TM4) cell lines were assessed by MTT assays, caspase activation, flow cytometry, TUNEL assay, Western blot, and cell cycle analysis. The exposure to different concentrations of terbufos ranging from 50 to 800 µM for 6 h caused significant death in all the used testicular cell lines. Terbufos increased reactive oxygen species (ROS) production, reduced mitochondrial membrane potential, and initiated apoptosis, which was confirmed by a dose-dependent increase in the number of TUNEL-positive apoptotic cells. Blocking ROS production by N-acetyl cysteine (NAC) protected GC-1 cells from terbufos-induced cell death. The results demonstrated that terbufos induces ROS, apoptosis, and DNA damage in testicular cell lines and it should be considered potentially hazardous to testis. Together, this study provided potential molecular mechanisms of terbufos-induced toxicity in testicular cells and suggests a possible protective measure. © 2015 Wiley Periodicals, Inc. Environ Toxicol 31: 1888-1898, 2016.

Adjacent disc and facet joint degeneration in young adults with low-grade spondylolytic spondylolisthesis: A magnetic resonance imaging study.

BACKGROUND/PURPOSE: Premature adjacent-level degeneration has been attributed to vertebral fusion, but spondylolisthesis has not been reported as a pathological factor responsible for the degeneration of adjacent disc and facet joint. We hypothesized that the degeneration of disc and facet joints in the adjacent levels is correlated with spondylolisthesis. METHODS: Magnetic resonance images of 35 symptomatic young adults (16-29 years old) with low-grade L5-S1 spondylolytic spondylolisthesis (Meyerding Grade 1 or 2) and 50 symptomatic young referents (20-29 years old) with L5-S1 disc herniation without spondylolisthesis were recruited to compare the differences between disc and facet-joint degenerations at the olisthetic and adjacent levels using the Mantel extension test. RESULTS: There were statistically significant degenerative changes of the discs and facet joints at the olisthetic and adjacent levels of patients with spondylolytic spondylolisthesis compared with the reference group. There is a trend that the disc and facet joints degenerate the most at the olisthetic level and become less affected at adjacent levels away from the lesion of pars defect. CONCLUSION: Low-grade spondylolytic spondylolisthesis was associated with significant degenerations of the disc and facet joints at olisthetic and adjacent levels in young adults.

The association between ambient air pollution exposure and connective tissue sarcoma risk: a nested case-control study using a nationwide population-based database.

A nationwide population-based database was utilized in a nested case-control study to explore the association between ambient air pollution exposure and the likelihood of developing connective tissue sarcoma. The study examined 280 cases of connective tissue sarcoma diagnosed between 2000 and 2012. A random sample of 1120 control subjects was selected from a subpopulation of claim records without a connective tissue sarcoma diagnosis in a 1:4 ratio. The control subjects were selected based on similar characteristics as the connective tissue sarcoma patients, including gender, birth year, and the year of diagnosis of the case group with medical records. Risk factors for connective tissue sarcoma were collected for analysis. Our data on exposure to air pollutants was collected from Taiwan's Air Quality Monitoring Network, which has been gathering air quality data from a growing network of sampling stations (now 76) throughout the country since 1997. It was discovered that the risk of connective tissue sarcoma was significantly increased by the Charlson comorbidity index (CCI), elevated levels of specific air pollution indices (e.g., total hydrocarbons (THC), fine particulate matter (PM2.5), and O3_8 (the annual mean of the daily maximum 8-h average concentration of O3), the High Pollutant Standards Index (hPSI) (the percentage of days in a given year in Taiwan where the PSI exceeds 100), and an insurable monthly wage over US$1100. Further investigation is needed to explore the involvement of these air pollutants in the formation of connective tissue sarcoma.

Clinical implications of linking microstructure, spatial biochemical, spatial biomechanical, and radiological features in ligamentum flavum degeneration.

Background: The ligamentum flavum (LF) degeneration is a critical factor in spinal stenosis, leading to nerve compression and pain. Even with new treatment options becoming available, it is vital to have a better understanding of LF degeneration to ensure the effectiveness of these treatments. Objective: This study aimed to provide insight into LF degeneration by examining the connections between various aspects of LF degeneration, including histology, microstructure, chemical composition, and biomechanics. Method: We analyzed 30 LF samples from 27 patients with lumbar vertebrae, employing magnetic resonance imaging (MRI) to link lumbar disc degeneration grades with fibrosis levels in the tissue. X-ray diffraction (XRD) analysis assessed microstructural alterations in the LF matrix component due to degeneration progression. Instrumented nanoindentation combined with Raman spectroscopy explored the spatial microbiomechanical and biochemical characteristics of the LF's ventral and dorsal regions. Results: Our outcomes revealed a clear association between the severity of LF fibrosis grades and increasing LF thickness. XRD analysis showed a rise in crystalline components and hydroxyapatite molecules with progressing degeneration. Raman spectroscopy detected changes in the ratio of phosphate, proteoglycan, and proline/hydroxyproline over the amide I band, indicating alterations in the extracellular matrix composition. Biomechanical testing demonstrated that LF tissue becomes stiffer and less extensible with increasing fibrosis. Discussion: Notably, the micro-spatial assessment revealed the dorsal side of the LF experiencing more significant mechanical stress, alongside more pronounced biochemical and biomechanical changes compared to the ventral side. Degeneration of the LF involves complex processes that affect tissue histology, chemical composition, and biomechanics. It is crucial to fully understand these changes to develop new and effective treatments for spinal stenosis. These findings can improve diagnostic accuracy, identify potential biomarkers and treatment targets, guide personalized treatment strategies, advance tissue engineering approaches, help make informed clinical decisions, and educate patients about LF degeneration.

Bone marrow edema in vertebral compression fractures: detection with dual-energy CT.

PURPOSE: To assess the use of the dual-energy computed tomographic (CT) virtual noncalcium technique in the evaluation of bone marrow edema in vertebral compression fractures. MATERIALS AND METHODS: This prospective study was approved by the institutional review board; informed consent was obtained from all patients. Sixty-three consecutive patients with 112 thoracic and/or lumbar vertebral compression fractures were studied between January 2011 and April 2012. All patients underwent both dual-energy CT (100 kV and Sn140 kV, where Sn indicates the use of a 0.4-mm tin filter) and magnetic resonance (MR) imaging. Dual-energy CT data were postprocessed by using a three-material decomposition algorithm for generating noncalcium images of the collapsed bodies. Two radiologists evaluated for the presence of abnormal attenuation alterations in the bone marrow by using color-coded maps and measured CT numbers on noncalcium grayscale images. Bone sclerosis and intravertebral air were evaluated with CT scans. MR images served as the reference standard. CT numbers were subjected to receiver operating characteristic curve analysis. RESULTS: MR imaging depicted 46 edematous and 66 nonedematous vertebral compression fractures. Eighty-two bodies were classified as having less than 50% sclerosis and/or air. Significant differences in noncalcium CT numbers between edematous and nonedematous vertebral compression fractures were found for both readers (P < .0001). CT numbers for the diagnosis of bone marrow edema on the basis of MR imaging revealed areas under the receiver operating characteristic curve of 0.799 and 0.841 for readers 1 and 2, respectively (P = .56). Use of a cutoff value of -80 to differentiate edematous vertebral bodies resulted in a sensitivity of 96.3%, specificity of 98.2%, and accuracy of 97.6% in the group of vertebral bodies with less than 50% sclerosis and/or air. CONCLUSION: Dual-energy CT virtual noncalcium images were able to depict bone marrow in the collapsed vertebral bodies, especially in those with less than 50% sclerosis and/or air.

MRI fluid sign is reliable in correlation with osteonecrosis after vertebral fractures: a histopathologic study.

INTRODUCTION: Magnetic resonance images (MRI) fluid sign and intravertebral vacuum phenomenon of the plain radiograph are considered as the characteristic radiological findings for vertebral osteonecrosis after spinal fractures. We aim to study the association between the radiological and histopathologic findings of vertebral osteonecrosis through the use of an open retrieval of specimens. MATERIALS AND METHODS: Twenty consecutive patients (54-84 years, mean 73 years) of unstable vertebral compression fractures treated with anterior corpectomy and fusion were included. All the images and pathologies were correlated, especially the histopathologic changes to the fluid sign and vacuum phenomenon. RESULTS: MRI fluid signs and the histopathologic findings of vertebral osteonecrosis were significantly correlated and both were noted in the first 5 months after injury. The power of the fluid sign in diagnosing vertebral osteonecrosis was better than that of the intravertebral vacuum phenomenon (diagnostic odds ratio 65 vs. 2, sensitivity 86 vs. 50%, specificity 100 vs. 67%). CONCLUSION: MRI fluid sign is more predictable to diagnose vertebral osteonecrosis in operative case, especially within the initial 5 months after injury.

Correlation of the degenerative stage of a disc with magnetic resonance imaging, chemical content, and biomechanical properties of the nucleus pulposus.

Intervertebral disc degeneration (IDD) is closely related to changes in the intervertebral disc (IVD) composition and the resulting viscoelastic properties. IDD is a severe condition because it decreases the disc's ability to resist mechanical loads. Our research aims to understand IDD at the cellular level, specifically the changes in the viscoelastic properties of the nucleus pulposus (NP), which are poorly understood. This study employed a system integrating nanoindentation with Raman spectrometry to correlate biomechanics with subtle changes in the biochemical makeup of the NP. The characterization was, in turn, correlated with the degenerative severity of IVD as assessed using magnetic resonance imaging (MRI) of different patients with spinal stenosis, degenerative spondylolisthesis, and degenerative scoliosis. It is shown that there is an increase in the crosslinking ratio in collagen, a reduction in proteoglycan, and a build-up of minerals upon the rise in the severity level of the disc damage in the NP. Assessment of mechanical characteristics reveals that the increasing disc degeneration makes the NP lose its elasticity, becoming more viscous. This shows that the tissue undergoes abnormalities in weight-bearing ability, which contributes to spinal instability. The correlation of the individual discs shows that grades III and IV have similarities in the changes of Amide I and III toward the storage modulus. In contrast, grades IV and V correlate with mineralization toward the storage modulus. Reduction of proteoglycan has the highest impact on the changes of the storage modulus in all grades of IDD. Connecting compositional alterations to IVD micromechanics at various degrees of degeneration expands our understanding of tissue behavior and provides critical insight into clinical diagnostics, treatment, and tissue engineering.

A methodology to evaluate different histological preparations of soft tissues: Intervertebral disc tissues study.

A tissue preparation method will inevitably alter the tissue content. This study aims to evaluate how different common sample preparation methods will affect the tissue morphology, biomechanical properties, and chemical composition of samples. The study focuses on intervertebral disc (IVD) tissue; however, it can be applied to other soft tissues. Raman spectroscopy synchronized with nanoindentation instrumentation was employed to investigate the compositional changes of IVD, specifically, nucleus pulposus (NP) and annulus fibrosus (AF), together with their biomechanical properties of IVD. These properties were examined through the following histological specimen types: fresh cryosection (control), fixed cryosection, and paraffin-embedded. The IVD tissue could be located using an optical microscope under three different preparation methods. Paraffin-embedded samples showed the most explicit details where the lamellae structure of AF could be identified. In terms of biomechanical properties, there was no significant difference between the fresh and fixed cryosection (p > 0.05). In contrast, the fresh cryosection and paraffin-embedded samples showed a significant difference (p < 0.05). It was also found that the tissue preparations affected the chemical content of the tissues and structure of the tissue, which are expected to contribute to biomechanical properties changes. Fresh cryosection and fixed cryosection samples are more promising to work with for biomechanical assessment in histological tissues. The findings fill essential gaps in the literature by providing valuable insight into the characteristics of IVD at the microscale. This study can also become a reference for a better approach to assessing the mechanical properties and chemical content of soft tissues at the microscale.

Anti-tumor effects of deubiquitinating enzyme inhibitor PR-619 in human chondrosarcoma through reduced cell proliferation and endoplasmic reticulum stress-related apoptosis.

Chondrosarcoma, a treatment-resistant cancer with limited therapeutic options, lacks significant advancements in treatment methods. However, PR-619, a novel inhibitor of deubiquitinating enzymes, has demonstrated anti-tumor effects in various malignancies. This study aimed to investigate the impact of PR-619 on chondrosarcoma both in vitro and in vivo. Two human chondrosarcoma cell lines, SW11353 and JJ012, were utilized. Cell viability was assessed using an MTT assay, while flow cytometry enabled the detection of apoptosis and cell cycle progression. Western blotting analyses were conducted to evaluate apoptosis, cell stress, and endoplasmic reticulum (ER) stress. Furthermore, the in vivo anti-tumor effects of PR-619 were examined using a xenograft mouse model. The results revealed that PR-619 induced cytotoxicity, apoptosis, and cell cycle arrest at the G0/G1 stage by activating caspases, PARP cleavage, and p21. Moreover, PR-619 increased the accumulation of polyubiquitinated proteins and ER stress by activating IRE1, GRP78, caspase-4, CHOP, and other cellular stress responses, including JNK activation. In vivo analysis demonstrated that PR-619 effectively inhibited tumor growth with minimal toxicity in the xenograft mouse model. These findings provide evidence of the anti-tumor effects and induction of cellular and ER stress by PR-619 in human chondrosarcoma, suggesting its potential as a novel therapeutic strategy for in human chondrosarcoma.

Development of Astaxanthin-Loaded Nanosized Liposomal Formulation to Improve Bone Health.

Astaxanthin is a xanthophyll carotenoid commonly found in marine organisms. Due to its super antioxidative ability, astaxanthin has been widely applied as a human nutraceutical supplement for health benefits. In order to enhance the bioavailability of astaxanthin, we used soybean phosphatidylcholine to encapsulate astaxanthin for liposomal formation. The physical properties of astaxanthin (asta)-loaded liposomes were determined by particle size, encapsulation efficiency and polydispersity index. The results revealed that the particle sizes of asta-loaded liposomes with various concentrations exhibited mean diameters in the range of 109 to 134 nm and had a narrow PDI value. As expected, the entrapment efficiency of liposomes loaded with a low concentration of astaxanthin (0.05 µg/mL) was 89%, and that was reduced to 29% for 1.02 µg/mL asta loading. Alizarin red staining and calcium content measurement showed that there was a significant reduction in calcium deposition for 7F2 osteoblasts treated with asta-loaded liposomes (0.25-1.02 µg/mL) in comparison with the cells treated with drug-free liposomes and mineralization medium (MM). Although liposomal formulation can reduce the cytotoxicity of astaxanthin and possess antioxidant, anti-inflammatory and anti-osteoclastogenic activities in RAW264.7 macrophages, asta-loaded liposomes with high concentrations may suppress ALP activity and mineralization level in 7F2 osteoblasts. Therefore, astaxanthin extract may be able to protect bones against oxidative stress and inflammation through liposomal formulation.

Mechanical Stretch Induced Osteogenesis on Human Annulus Fibrosus Cells through Upregulation of BMP-2/6 Heterodimer and Activation of P38 and SMAD1/5/8 Signaling Pathways.

Degenerative disc disease (DDD) is an important cause of low back pain. Repetitive tensile stress from the daily motion of the spine predisposes it to injury of the annulus fibrosus (AF) which causes IVD degeneration. This study aims to determine the causal relationship between mechanical stretch and osteogenesis in the AF cells of IVD as affected by bone morphogenic proteins (BMPs), specifically BMP-2/6 heterodimers. Our results found that 15% tensile stress (high cyclic stretching, HCS) may induce the expression of osteogenesis-related markers (Runx2, osterix) by upregulating BMP-2/6 heterodimeric ligands and their receptors on the human AF cell line. HCS also induced transient phosphorylation of p38 mitogen-activated protein (MAP) kinase and SMAD1/5/8. Neutralizing antibodies to the BMP-2/6 receptor (ALK3) blocked the expression of Runx2 and osterix, as well as the phosphorylation of p38 and SMAD1/5/8. In addition, treatment with a p38 MAPK inhibitor (SB203580) or siRNA to neutralize the effects of SMAD1/5/8 suppressed tensile stress-induced Runx2 and osterix expression. Mechanical stretching induces activation of p38 MAP kinase and SMAD1/5/8 signaling pathways, followed by the upregulation of BMP-2/6 heterodimer expression, thereby stimulating osteogenic Runx2 and osterix expression on AF cells. HCS may accelerate the progression of IVD degeneration by promoting an osteogenic response.

Optimization of Spinal Reconstructions for Thoracolumbar Burst Fractures to Prevent Proximal Junctional Complications: A Finite Element Study.

The management strategies of thoracolumbar (TL) burst fractures include posterior, anterior, and combined approaches. However, the rigid constructs pose a risk of proximal junctional failure. In this study, we aim to systemically evaluate the biomechanical performance of different TL reconstruction constructs using finite element analysis. Furthermore, we investigate the motion and the stress on the proximal junctional level adjacent to the constructs. We used a T10-L3 finite element model and simulated L1 burst fracture. Reconstruction with posterior instrumentation (PI) alone (U2L2 and U1L1+(intermediate screw) and three-column spinal reconstruction (TCSR) constructs (U1L1+PMMA and U1L1+Cage) were compared. Long-segment PI resulted in greater global motion reduction compared to constructs with short-segment PI. TCSR constructs provided better stabilization in L1 compared to PI alone. Decreased intradiscal and intravertebral pressure in the proximal level were observed in U1L1+IS, U1L1+PMMA, and U1L1+Cage compared to U2L2. The stress and strain energy of the pedicle screws decreased when anterior reconstruction was performed in addition to PI. We showed that TCSR with anterior reconstruction and SSPI provided sufficient immobilization while offering additional advantages in the preservation of physiological motion, the decreased burden on the proximal junctional level, and lower risk of implant failure.

Sitting Posture during Prolonged Computer Typing with and without a Wearable Biofeedback Sensor.

Prolonged sitting combined with an awkward posture might contribute to the increased risks of developing spinal pain. Maintaining an upright sitting posture is thus often suggested, especially nowadays when people spend longer periods in the sitting posture for occupational or leisure activities. Many types of assistive devices are commercially available to help computer users maintain an upright sitting posture. As the technology advances, wearable sensors that use microelectromechanical technology are designed to provide real-time biofeedback and promote adjusting posture actively. However, whether such wearable biofeedback sensors could assist adjusting sitting posture in computer users during prolonged typing remains unknown. This study aimed to investigate the effects of a wearable biofeedback sensor on maintaining an upright sitting posture. Twenty-one healthy young adults were recruited and performed a 1-h computer typing task twice, with and without using the active biofeedback device. The sagittal spinal posture during computer typing was measured using a three-dimensional motion analysis system. Using the wearable biofeedback sensor significantly decreased the neck flexion (p < 0.001), thoracic kyphotic (p = 0.033), and pelvic plane (p = 0.021) angles compared with not using the sensor. Computer users and sedentary workers may benefit from using wearable biofeedback sensors to actively maintain an upright sitting posture during prolonged deskwork.

Biomechanical analysis of the lumbar spine on facet joint force and intradiscal pressure--a finite element study.

BACKGROUND: Finite element analysis results will show significant differences if the model used is performed under various material properties, geometries, loading modes or other conditions. This study adopted an FE model, taking into account the possible asymmetry inherently existing in the spine with respect to the sagittal plane, with a more geometrically realistic outline to analyze and compare the biomechanical behaviour of the lumbar spine with regard to the facet force and intradiscal pressure, which are associated with low back pain symptoms and other spinal disorders. Dealing carefully with the contact surfaces of the facet joints at various levels of the lumbar spine can potentially help us further ascertain physiological behaviour concerning the frictional effects of facet joints under separate loadings or the responses to the compressive loads in the discs. METHODS: A lumbar spine model was constructed from processes including smoothing the bony outline of each scan image, stacking the boundary lines into a smooth surface model, and subsequent further processing in order to conform with the purpose of effective finite element analysis performance. For simplicity, most spinal components were modelled as isotropic and linear materials with the exception of spinal ligaments (bilinear). The contact behaviour of the facet joints and changes of the intradiscal pressure with different postures were analyzed. RESULTS: The results revealed that asymmetric responses of the facet joint forces exist in various postures and that such effect is amplified with larger loadings. In axial rotation, the facet joint forces were relatively larger in the contralateral facet joints than in the ipsilateral ones at the same level. Although the effect of the preloads on facet joint forces was not apparent, intradiscal pressure did increase with preload, and its magnitude increased more markedly in flexion than in extension and axial rotation. CONCLUSIONS: Disc pressures showed a significant increase with preload and changed more noticeably in flexion than in extension or in axial rotation. Compared with the applied preloads, the postures played a more important role, especially in axial rotation; the facet joint forces were increased in the contralateral facet joints as compared to the ipsilateral ones at the same level of the lumbar spine.