siRNA incorporated in slow-release injectable hydrogel continuously silences DDIT4 and regulates nucleus pulposus cell pyroptosis through the ROS/TXNIP/NLRP3 axis to alleviate intervertebral disc degeneration.

Aims: In this investigation, we administered oxidative stress to nucleus pulposus cells (NPCs), recognized DNA-damage-inducible transcript 4 (DDIT4) as a component in intervertebral disc degeneration (IVDD), and devised a hydrogel capable of conveying small interfering RNA (siRNA) to IVDD. Methods: An in vitro model for oxidative stress-induced injury in NPCs was developed to elucidate the mechanisms underlying the upregulation of DDIT4 expression, activation of the reactive oxygen species (ROS)-thioredoxin-interacting protein (TXNIP)-NLRP3 signalling pathway, and nucleus pulposus pyroptosis. Furthermore, the mechanism of action of small interfering DDIT4 (siDDIT4) on NPCs in vitro was validated. A triplex hydrogel named siDDIT4@G5-P-HA was created by adsorbing siDDIT4 onto fifth-generation polyamidoamine (PAMAM) dendrimer using van der Waals interactions, and then coating it with hyaluronic acid (HA). In addition, we established a rat puncture IVDD model to decipher the hydrogel's mechanism in IVDD. Results: A correlation between DDIT4 expression levels and disc degeneration was shown with human nucleus pulposus and needle-punctured rat disc specimens. We confirmed that DDIT4 was responsible for activating the ROS-TXNIP-NLRP3 axis during oxidative stress-induced pyroptosis in rat nucleus pulposus in vitro. Mitochondria were damaged during oxidative stress, and DDIT4 contributed to mitochondrial damage and ROS production. In addition, siDDIT4@G5-P-HA hydrogels showed good delivery activity of siDDIT4 to NPCs. In vitro studies illustrated the potential of the siDDIT4@G5-P-HA hydrogel for alleviating IVDD in rats. Conclusion: DDIT4 is a key player in mediating pyroptosis and IVDD in NPCs through the ROS-TXNIP-NLRP3 axis. Additionally, siDDIT4@G5-P-HA hydrogel has been found to relieve IVDD in rats. Our research offers an innovative treatment option for IVDD.

Numerical study of pedicle screw construction and locking compression plate fixation in posterior pelvic ring injuries: Analyzed by finite element method.

BACKGROUND: The aim of this study was to compare the biomechanical performance of pedicle screw construction and locking compression plate fixation in posterior pelvic ring injuries analyzed by finite element method. METHODS: A 3-dimensional finite element model of the spine-pelvis-femur complex with ligaments was reconstructed from computed tomography images. An unstable posterior pelvic ring injury was created, which was fixed with a pedicle screw construction or locking compression plate. A follower load of 400 N was applied to the upper surface of the vertebrae to simulate the upper body weight, while the ends of the proximal femurs were fixed. The construct stiffness, the maximum vertical displacement, the maximum posterior displacement, the maximum right displacement, and the overall maximum displacement of the sacrum, and stress distributions of the implants and pelvises were assessed. RESULTS: The construct stiffness of the pedicle screw model (435.14 N/mm) was 2 times that of the plate model (217.01 N/mm). The maximum vertical displacement, the maximum posterior displacement, the maximum right displacement, and the overall maximum displacement of the sacrum in the pedicle screw model were smaller than those in the plate model (0.919, 1.299, 0.259, and 1.413 mm in the pedicle screw model, and 1.843, 2.300, 1.053, and 2.895 mm in the plate model, respectively). The peak stresses of the implant and pelvis in the pedicle screw model decreased by 80.4% and 25% when compared with the plate model (44.57 and 34.48 MPa in the pedicle screw model, and 227.47 and 45.97 MPa in the plate model, respectively). CONCLUSION: The study suggested that the pedicle screw construction could provide better fixation stability than the locking compression plate and serves as the recommended fixation method for the treatment of posterior pelvic ring injuries.

Expert consensus on Prospective Precision Diagnosis and Treatment Strategies for Osteoporotic Fractures.

Osteoporotic fractures are the most severe complications of osteoporosis, characterized by poor bone quality, difficult realignment and fixation, slow fracture healing, and a high risk of recurrence. Clinically managing these fractures is relatively challenging, and in the context of rapid aging, they pose significant social hazards. The rapid advancement of disciplines such as biophysics and biochemistry brings new opportunities for future medical diagnosis and treatment. However, there has been limited attention to precision diagnosis and treatment strategies for osteoporotic fractures both domestically and internationally. In response to this, the Chinese Medical Association Orthopaedic Branch Youth Osteoporosis Group, Chinese Geriatrics Society Geriatric Orthopaedics Committee, Chinese Medical Doctor Association Orthopaedic Physicians Branch Youth Committee Osteoporosis Group, and Shanghai Association of Integrated Traditional Chinese and Western Medicine Osteoporosis Professional Committee have collaborated to develop this consensus. It aims to elucidate emerging technologies that may play a pivotal role in both diagnosis and treatment, advocating for clinicians to embrace interdisciplinary approaches and incorporate these new technologies into their practice. Ultimately, the goal is to improve the prognosis and quality of life for elderly patients with osteoporotic fractures.

A novel degradable PCL/PLLA strapping band for internal fixation of fracture.

Early fracture fixation is the critical factor in fracture healing. Common internal fracture implants are made of metallic materials, which often affects the imaging quality of CT and MRI. Most patients will choose secondary surgery to remove the internal fixation implants, which causes secondary damage to them. The development of new degradable internal fracture implants has attracted more and more attention from orthopedic surgeons and researchers. Based on these problems, we improved the various properties of medical grade polycaprolactone (PCL) by adding poly(L-lactide) (PLLA). We produced PCL/PLLA strapping bands with different mass ratios by injection molding. We compared the mechanical properties, degradation properties, cell biocompatibility, bone marrow mesenchymal stem cells (BMSCs) adhesion, proliferation, osteogenic differentiation and fracture fixation effect of these strapping bands. The results showed that the tensile strength and yield force of the strapping bands increased with the increase of the content of PLLA. The addition of PLLA could significantly improve the mechanical strength in the early stage and accelerate the degradation rate of the strapping band. PCL/PLLA (80/20) strapping band had no significant cytotoxicity toward rBMSCs and could promote osteogenic differentiation of rBMSCs. The strapping band could ensure femoral fracture healing of beagles in 3 months and didn't cause damage to the surrounding tissues and main organs. This study will provide some new insights into the biodegradable products of PCL/PLLA blends for internal fixation of fracture. We produced novel degradable PCL/PLLA strapping bands with different mass ratios by injection molding. We tested the biological safety of the prepared internal fixation strapping bands for fracture, such as cell experiment in vitro and animal experiment, and studied the degradation behavior in vitro. The strapping bands could ensure femoral fracture healing of beagles. This study will provide some new insights into the biodegradable products of PCL/PLLA blends for internal fixation of fracture. A Immunofluorescence staining of rBMSCs (live cells: green; dead cells: red). B Young's modulus change curve during strapping bands degradation. C The implantation process of strapping bands. D Micro-CT images of the beagle's fracture recovery after the operation.

NFATc1-mediated expression of SLC7A11 drives sensitivity to TXNRD1 inhibitors in osteoclast precursors.

Excess osteoclast activity is found in many bone metabolic diseases, and inhibiting osteoclast differentiation has proven to be an effective strategy. Here, we revealed that osteoclast precursors (pre-OCs) were more susceptible to thioredoxin reductase 1 (TXNRD1) inhibitors than bone marrow-derived monocytes (BMDMs) during receptor activator of nuclear factor kappa B ligand (RANKL)-mediated osteoclastogenesis. Mechanistically, we found that nuclear factor of activated T-cells 1 (NFATc1) upregulated solute carrier family 7 member 11 (SLC7A11) expression through transcriptional regulation during RANKL-induced osteoclastogenesis. During TXNRD1 inhibition, the rate of intracellular disulfide reduction is significantly reduced. Increased cystine transport leads to increased cystine accumulation, which leads to increased cellular disulfide stress and disulfidptosis. We further demonstrated that SLC7A11 inhibitors and treatments that prevent disulphide accumulation could rescue this type of cell death, but not the ferroptosis inhibitors (DFO, Ferro-1), the ROS scavengers (Trolox, Tempol), the apoptosis inhibitor (Z-VAD), the necroptosis inhibitor (Nec-1), or the autophagy inhibitor (CQ). An in vivo study indicated that TXNRD1 inhibitors increased bone cystine content, reduced the number of osteoclasts, and alleviated bone loss in an ovariectomized (OVX) mouse model. Together, our findings demonstrate that NFATc1-mediated upregulation of SLC7A11 induces targetable metabolic sensitivity to TXNRD1 inhibitors during osteoclast differentiation. Moreover, we innovatively suggest that TXNRD1 inhibitors, a classic drug for osteoclast-related diseases, selectively kill pre-OCs by inducing intracellular cystine accumulation and subsequent disulfidptosis.

Finite Element Analysis of Channel Screw and Conventional Plate Technique in Tile B2 Pelvic Fracture.

OBJECTIVE: This study aims to analyze the biomechanical characteristics of tile B2 pelvic fractures using finite element analysis when the superior ramus of the pubis was fixed by a plate or hollow screws in standing and sitting positions, respectively. METHODS: A three-dimensional digital model of the tile B2 pelvic fracture was obtained by CT scanning the patient. The main ligament structure was then reconstructed based on the anatomical characteristics to create a finite element model of the tile B2 pelvic fracture. The posterior pelvic ring was fixed by sacroiliac joint screws, while the anterior ring injury of the superior ramus of the pubis was fixed by plates and hollow compression screws, respectively. The degrees of freedom of the bilateral acetabulum or two sides of the ischial tuberosity were constrained in the two models. A vertical load of 600 N was applied to the upper surface of the sacrum to measure the displacement and stress distribution of the pelvis in the standing and sitting positions. RESULTS: The displacement distribution of both the healthy and the affected side of the pelvis was relatively uniform in both the plate group and the hollow screw group according to the finite element simulation results. The maximum displacement value in the sitting position was greater than the standing position, and the maximum displacement value of the hollow screw fixation was greater than that of the plate fixation. In the four groups of fixation models, the maximum displacement value of the pelvis in the hollow screw sitting position group was 1616.80 × 10-3 mm, which was greater than that of the other three groups, and in this group the total displacement value of the hollow screw in the anterior ring was 556.31 × 10-3 mm. The stress distribution of the pelvis in the various models was similar in the four groups of models, in which the maximum stress of the pelvis in the hollow screw sitting position group was the largest, which was 201.33 MPa, while the maximum stress in the standing position was 149.85 MPa greater than that in the sitting position of the hollow screw fixation. CONCLUSION: The anterior ring of patients with Tile B2 pelvic fractures fixed with hollow screws or plates in both standing and sitting positions can achieve satisfactory biomechanical results with significant safety margins for plates and screws.

Expert consensus on the bone repair strategy for osteoporotic fractures in China.

Osteoporotic fractures, also known as fragility fractures, are prevalent in the elderly and bring tremendous social burdens. Poor bone quality, weak repair capacity, instability, and high failure rate of internal fixation are main characteristics of osteoporotic fractures. Osteoporotic bone defects are common and need to be repaired by appropriate materials. Proximal humerus, distal radius, tibia plateau, calcaneus, and spine are common osteoporotic fractures with bone defect. Here, the consensus from the Osteoporosis Group of Chinese Orthopaedic Association concentrates on the epidemiology, characters, and management strategies of common osteoporotic fractures with bone defect to standardize clinical practice in bone repair of osteoporotic fractures.

Mesenchymal stem cells encapsulation in chitosan and carboxymethyl chitosan hydrogels to enhance osteo-differentiation.

BACKGROUND: Recently biomaterials utilized for designing scaffolds in tissue engineering are not cost-effective and eco-friendly. As a result, we design and develop biocompatible and bioactive hydrogels for osteo-tissue regeneration based on the natural polysaccharide chitosan. Three distinct hydrogel components were used for this. METHODS: Hydrogels networks were created using chitosan 2% (CTS 2%), carboxymethyl chitosan 2% (CMC 2%), and 50:50 mixtures of CTS and CMC (CTS/CMC 50:50). Furthermore, scanning electron microscopy (SEM), Fourier transforms infrared spectroscopy (FTIR), degradation, and swelling behavior of design hydrogels were studied. Also, the cytocompatibility and osteo-differentiation potency were examined by encapsulating mesenchymal stem cells derived from adipose tissue (AMSCs) on the designed hydrogels. RESULTS: According to the findings, our results showed an acceptable pore structure, functional groups, and degradation rate of the designed hydrogels for in vitro evaluation. In addition, employing CMC instead of CTS or adding 50% CMC to the hydrogel component could improve the hydrogel's osteo-bioactivity without the use of external osteogenic differentiation agents. CONCLUSION: The CMC-containing hydrogel not only caused early osteogenesis but also accelerated differentiation to the maturity phase of osteoblasts.

The effects of process parameters on polydopamine coatings employed in tissue engineering applications.

The biomaterials' success within the tissue engineering field is hinged on the capability to regulate tissue and cell responses, comprising cellular adhesion, as well as repair and immune processes' induction. In an attempt to enhance and fulfill these biomaterials' functions, scholars have been inspired by nature; in this regard, surface modification via coating the biomaterials with polydopamine is one of the most successful inspirations endowing the biomaterials with surface adhesive properties. By employing this approach, favorable results have been achieved in various tissue engineering-related experiments, a significant one of which is the more rapid cellular growth observed on the polydopamine-coated substrates compared to the untreated ones; nonetheless, some considerations regarding polydopamine-coated surfaces should be taken into account to control the ultimate outcomes. In this mini-review, the importance of coatings in the tissue engineering field, the different types of surfaces requiring coatings, the significance of polydopamine coatings, critical factors affecting the result of the coating procedure, and recent investigations concerning applications of polydopamine-coated biomaterials in tissue engineering are thoroughly discussed.

Injectable light-assisted thermo-responsive methylcellulose-sodium humate hydrogel proposed for photothermal ablation and localized delivery of cisplatin.

This study aimed to develop injectable light-assisted thermo-responsive methylcellulose hydrogels filled with sodium humate, which were proposed for photothermal ablation and localized cisplatin delivery. Sodium humate converts light energy from laser beams into thermal energy, which causes methylcellulose to gel, thereby controlling the release of chemotherapy agents. Meanwhile, light emission causes to the photothermal ablation of tumor cells. For determining the optimal production conditions, different concentrations of sodium humate and light emission times were investigated. Results show that hydrogel uniformity is highly dependent on variables. An increase in sodium humate concentration and emission time resulted in a slight reduction in swelling ratio and an increase in durability. According to the simulation conditions, the cisplatin release profile was consistent with a non-Fickian mechanism with a predominant erosion contribution. In conjugation with increasing light emission time and sodium humate content, the storage modulus and viscosity increased, demonstrating hydrogel's sol-gel transition and long-lasting durability. The intrinsic fluorescence spectroscopy study revealed that the hydrogel-model protein complex empowered hydrogel bio-performance. Laser emission and cisplatin release synergistically reduced the number of viable osteosarcoma cell lines, suggesting the possibility of tumor ablation. This study describes the potential of simultaneous photothermal therapy and chemotherapy in osteosarcoma treatment, laying the groundwork for future preclinical and clinical trials.

The Dibenzyl Isoquinoline Alkaloid Berbamine Ameliorates Osteoporosis by Inhibiting Bone Resorption.

Postmenopausal osteoporosis (PMOP) is a kind of primary osteoporosis that is characterized by decreased bone density and strength. Berbamine is a nonbasic quaternary benzylisoquinoline plant alkaloid that has been widely used in the clinic to treat leukopenia in China. We found that berbamine inhibited RANKL-induced osteoclastogenesis of bone marrow-derived macrophages (BMMs) in vitro, which mainly occurred in the middle phase and late phase. The gene and protein expression levels of osteoclast-related molecules, including CTSK, MMP-9, NFATc1, CD44 and DC-STAMP, were also downregulated by berbamine. In vivo, we treated PMOP mice with berbamine for 8 weeks and found that the extent of osteoporosis was alleviated significantly according to micro-CT scanning, hematoxylin-eosin staining, DC-STAMP immunohistochemical staining and TRAP immunohistochemical staining in the distal femurs of the mice. Our findings demonstrate that berbamine has an inhibitory effect on the osteoclastogenesis of BMMs and can prevent bone loss after ovariectomy in vivo. This study provides evidence that berbamine is a potential drug for the prevention and treatment of PMOP.

Perioperative Protocol of Ankle Fracture and Distal Radius Fracture Based on Enhanced Recovery after Surgery Program: A Multicenter Prospective Clinical Controlled study.

Background: The enhanced recovery after surgery (ERAS) program is aimed to shorten patients' recovery process and improve clinical outcomes. This study aimed to compare the outcomes between the ERAS program and the traditional pathway among patients with ankle fracture and distal radius fracture. Methods: This is a multicenter prospective clinical controlled study consisting of 323 consecutive adults with ankle fracture from 12 centers and 323 consecutive adults with distal radial fracture from 13 centers scheduled for open reduction and internal fixation between January 2017 and December 2018. According to the perioperative protocol, patients were divided into two groups: the ERAS group and the traditional group. The primary outcome was the patients' satisfaction of the whole treatment on discharge and at 6 months postoperatively. The secondary outcomes include delapsed time between admission and surgery, length of hospital stay, postoperative complications, functional score, and the MOS item short form health survey-36. Results: Data describing 772 patients with ankle fracture and 658 patients with distal radius fracture were collected, of which 323 patients with ankle fracture and 323 patients with distal radial fracture were included for analysis. The patients in the ERAS group showed higher satisfaction levels on discharge and at 6 months postoperatively than in the traditional group (P < 0.001). In the subgroup analysis, patients with distal radial fracture in the ERAS group were more satisfied with the treatment (P=0.001). Furthermore, patients with ankle fracture had less time in bed (P < 0.001) and shorter hospital stay (P < 0.001) and patients with distal radial fracture received surgery quickly after being admitted into the ward in the ERAS group than in the traditional group (P=0.001). Conclusions: Perioperative protocol based on the ERAS program was associated with high satisfaction levels, less time in bed, and short hospital stay without increased complication rate and decreased functional outcomes.

Molybdenum disulfide nanosheet/polyimide composites with improved tribological performances, surface properties, antibacterial effects and osteogenesis for facilitating osseointegration.

Polymeric biocomposites display some advantages over metal or ceramic biomaterials, and are regarded as a promising candidate for artificial joint application. Herein, molybdenum disulfide (MD) nanosheets were prepared and incorporated into polyimide (PI) to form MD/PI composites with a MD content of 20 wt% (PM20) and 40 wt% (PM40). The results revealed that incorporation of MD nanosheets obviously improved the tribological performances, surface properties (e.g., roughness, wettability and surface energy) and protein absorption of the composites, which enhanced with the increase of MD content. In addition, the composites containing MD nanosheets exhibited antibacterial effects, and the antibacterial effects of PM40 were higher than those of PM20 and PI. PM40 significantly stimulated the cellular responses of rat bone mesenchymal stem cells in vitro, which was better than PM20 and PI. Furthermore, PM40 remarkably accelerated osteogenesis and osseointegration in vivo, which was better than PM20 and PI. In summary, the MD content in composites played pivotal roles in improving not only tribological performances, surface properties, antibacterial effects and cellular response in vitro but also osteogenesis and osseointegration in vivo. As a result, PM40 with high MD content exhibited excellent osteogenic bioactivity and antibacterial effects, which would have great potential for artificial joint applications.

Ring-Pins combined with cable cerclage for the fixation of displaced inferior patellar pole fractures.

Objective: The study aimed to present the clinical results and complication rates of ring-pins with cable cerclage for treating the inferior pole of patella fracture. Method: A study that retrospectively reviewed consecutive patients of the displaced inferior pole of patella fracture (AO/OTA 34-A1) operated with a ring-pin tension band using cable cerclage between October 2015 and October 2017 was performed. The duration of surgery, motion range of the knee, function outcomes, and complications were recorded. Results: The average follow-up of 31 patients was 21 months. The mean operation time was 50 min. Fractures in all 31 patients healed at a mean duration of 8 weeks. There was no infection, no withdrawing of ring-pins, no implant breakage, and no loss of fracture reduction. The mean range of motion was 120°, and no patient complained of implant irritation at the final follow-up. The average Bostman score was 29.0 points, and 28 patients graded clinical outcomes excellent and 3 patients graded clinical outcomes good at the last follow-up. Conclusions: Ring-pin combined with cable cerclage for treating the displaced inferior pole of patellar fracture is simple, and the postoperative internal fixation-related complication rate is low. It is a good choice for treating the displaced inferior pole of the patellar fracture.

Polydopamine Coating-Mediated Immobilization of BMP-2 on Polyethylene Terephthalate-Based Artificial Ligaments for Enhanced Bioactivity.

Background/objectives: Polyethylene terephthalate (PET)-based artificial ligaments are one of the most commonly used grafts in anterior cruciate ligament (ACL) reconstruction surgery. However, the lack of favorable hydrophilicity and cell attachment for PET highly impeded its widespread application in clinical practice. Studies found that surface modification on PET materials could enhance the biocompatibility and bioactivity of PET ligaments. In this study, we immobilized bone morphogenetic protein-2 (BMP-2) on the surface of PET ligaments mediated by polydopamine (PDA) coating and investigated the bioactivation and graft-to-bone healing effect of the modified grafts in vivo and in vitro. Methods: In this study, we prepared the PDA coating and subsequent BMP-2-immobilized PET artificial ligaments. Scanning electron microscopy (SEM) was used to analyze the morphological changes of the modified grafts. In addition, the surface wettability properties of the modified ligaments, amount of immobilized BMP 2, and the release of BMP-2 during a dynamic period up to 28 days were tested. Then, the attachment and proliferation of rat bone mesenchymal stem cells (rBMSCs) on grafts were examined by SEM and Cell Counting Kit-8 (CCK-8) assay, respectively. Alkaline phosphatase (ALP) assay, RT-PCR, and Alizarin Red S staining were performed to test the osteoinduction property. For in vivo experiments, an extra-articular graft-to-bone healing model in rabbits was established. At 8 weeks after surgery, biomechanical tests, micro-CT, and histological staining were performed on harvested samples. Results: A surface morphological analysis verified the success of the PDA coating. The wettability of the PET artificial ligaments was improved, and more than 80% of BMP-2 stably remained on the graft surface for 28 days. The modified grafts could significantly enhance the proliferation, attachment, as well as expression of ALP and osteogenic-related genes, which demonstrated the favorable bioactivity of the grafts immobilized with BMP-2 in vitro. Moreover, the grafts immobilized with BMP-2 at a concentration of 138.4 ± 10.6 ng/cm2 could highly improve the biomechanical properties, bone regeneration, and healing between grafts and host bone after the implantation into the rabbits compared with the PDA-PET group or the PET group. Conclusion: The immobilization of BMP-2 mediated by polydopamine coating on PET artificial ligament surface could enhance the compatibility and bioactivity of the scaffolds and the graft-to-bone healing in vivo.

Injectable Mussel-Inspired highly adhesive hydrogel with exosomes for endogenous cell recruitment and cartilage defect regeneration.

In the early stage of osteoarthritis (OA), cartilage degradation in the surface region leads to superficial cartilage defect. However, enhancing the regeneration of cartilage defect remains a great challenge for existing hydrogel technology because of the weak adhesion to wet tissue. In the present study, an injectable mussel-inspired highly adhesive hydrogel with exosomes was investigated for endogenous cell recruitment and cartilage defect regeneration. The hydrogel with high bonding strength to the wet surface was prepared using a crosslinked network of alginate-dopamine, chondroitin sulfate, and regenerated silk fibroin (AD/CS/RSF). Compared with commercial enbucrilate tissue adhesive, the AD/CS/RSF hydrogel provided a comparative lap shear strength of 120 kPa, with a similar gelation time and a higher capacity for maintaining adhesive strength. The AD/CS/RSF/EXO hydrogel with encapsulated exosomes recruited BMSCs migration and inflation, promoted BMSCs proliferation and differentiation. Most importantly, the AD/CS/RSF/EXO hydrogel accelerated cartilage defect regeneration in situ, and extracellular matrix remodeling after injection in rat patellar grooves. The exosomes released by the hydrogels could recruit BMSCs into the hydrogel and neo-cartilage via the chemokine signaling pathway. Our findings reveal an injectable and adhesive hydrogel for superficial cartilage regeneration, which is a promising approach for minimally treating cartilage defect with arthroscopic assistance.

Tantalum oxide submicro-particles into microporous coating on polyimide possessing antibacterial property and inducing cellular response for orthopedic application.

Surface characteristics of the biomaterials have significant effects on response of osteoblast and formation of new bone tissue. In this study, to improve the bio-performance of polyimide (PI) as an implantable material for bone substitute, concentrated sulfuric acid suspension with tantalum (V) oxide (vTO) submicro-particles of 10w% (PIST10) and 15w% (PIST15) was utilized to modify PI surface. After sulfonation, microporous coatings including vTO particles were created on PI (PIST10 and PIST15) while microporous coating without vTO particles was also created on PI (PIS). Results showed that surface roughness, hydrophilicity and protein adsorption of PIST15 was remarkably higher than PIST10 and PIS. Furthermore, after soaking into simulated body fluid (SBF), no apatite mineralization on PIS was found, while PIST15 with high vTO content exhibited better apatite mineralization compared with PIST10. Moreover, PIS showed low antibacterial property, while PIST15 with high vTO content revealed better antibacterial property compared with PIST10. In addition, cellular response (such as adhesion, proliferation and alkaline phosphatase activity) of bone marrow stromal cells (BMSC) of rat to PIST15 was higher than PIST10 and PIS. In conclusion, the microporous coating of PIST15 including vTO submicro-particles possessed good antibacterial property and bioactivity, which significantly promoted the responses of BMSC. Therefore, PIST15 has potential application prospects for bone substitute.

Hypoxia inhibits RANKL-induced ferritinophagy and protects osteoclasts from ferroptosis.

Ferroptosis is a new form of regulated cell death. Several studies have demonstrated that ferroptosis was involved in multiple diseases. However, the precise role of ferroptosis in osteoporosis remains unclear. Here, we demonstrated that ferroptosis was involved in osteoclasts over the course of RANKL-induced differentiation, and it was induced by iron-starvation response and ferrintinophagy. Mechanistically, under normoxia but not hypoxia, ferroptosis could be induced due to iron-starvation response (increased transferrin receptor 1, decreased ferritin) followed by RANKL stimulation, and this was attributed to the down-regulation of aconitase activity. We further investigated intracellular iron homeostasis and found that ferritinophagy, a process initiated by FTH-NCOA4 complex autophagosome degradation, was activated followed by RANKL stimulation under normoxia. Interestingly, these processes could not be observed under hypoxia. Moreover, we demonstrated that HIF-1α contributed to the decrease of ferritinophagy and autophagy flux under hypoxia. Additionally, HIF-1α impair autophagy flux via inhibition of autophagosome formation under hypoxia in BMDMs. In vivo study, we indicated that HIF-1α specific inhibitor 2ME2 prevent OVX bone loss. In conclusion, our study comprehensively investigated the role of ferroptosis in osteoclasts in vitro and in vivo, and innovatively suggested that targeting HIF-1α and ferritin thus inducing ferroptosis in osteoclasts could be an alternative in treatment of osteoporosis.

Oxyresveratrol induces apoptosis and inhibits cell viability via inhibition of the STAT3 signaling pathway in Saos­2 cells.

Oxyresveratrol (ORES) is a natural phenolic compound with multiple biological functions including antioxidation, anti­inflammation and neuroprotection; however, the inhibitory effect of ORES on osteosarcoma remains largely unknown. The present study aimed to determine the effects of ORES on osteosarcoma cell Saos­2. Cell Counting Kit­8 assay was performed to detect Soas­2 cell viability. Annexin­FITC/PI staining and JC­1 staining were used to measure cell apoptosis and the change of mitochondrial membrane potential. In addition, western blotting was conducted to determine the expression levels of apoptotic proteins and the phosphorylation of STAT3. It was found that ORES inhibited cell viability and induced apoptosis of osteosarcoma Saos­2 cells in a concentration­dependent manner. In addition, ORES increased the expression levels of apoptotic proteases caspase­9 and caspase­3 and reduced mitochondrial membrane potential. In response to ORES treatment, the expression levels of pro­apoptotic proteins, Bad and Bax, were enhanced, whereas those of anti­apoptotic proteins, Bcl­2 and Bcl­xL, were reduced. In addition, the phosphorylation of STAT3 was attenuated in Saos­2 cells after treatment with ORES. Inhibition of cell viability and apoptosis induction by ORES were rescued by enhancement of STAT3 activation upon treatment with IL­6. Collectively, the present study indicated that ORES induced apoptosis and inhibited cell viability, which may be associated with the inhibition of STAT3 activation; thus, ORES represents a promising agent for treating osteosarcoma.

Schisandrin A restrains osteoclastogenesis by inhibiting reactive oxygen species and activating Nrf2 signalling.

OBJECTIVES: Intracellular reactive oxygen species (ROS) induced by receptor activator of NF-kB ligand (RANKL) has been proven to be a critical factor in the development of osteoclasts. This study aimed to prove that schisandrin A (Sch), a novel anti-oxidant compound, is able to suppress osteoclastogenesis and prevent bone loss in ovariectomized (OVX) mice by suppressing ROS via nuclear factor erythroid 2-related factor (Nrf2). MATERIAL AND METHODS: Micro-CT was used to detect bone formation. The effects of Sch on receptor activator of nuclear factor-κB (NF-κB) ligand (RANKL)-induced reactive oxygen species (ROS) were measured by dihydroethidium (DHE) staining in vivo and 2',7'-dichlorodihydrofluorescein diacetate (DCFH-DA) staining in vitro. Immunofluorescence staining was used to detect the expression of Nrf2 in vivo. siRNA was used to evaluate the effect of Nrf2 in osteoclastogenesis. RESULTS: Sch suppresses RANKL-induced ROS production by regulating nuclear factor erythroid 2-related factor (Nrf2) in vitro and vivo. Mechanistically, Sch enhances the expression of Nrf2 by regulating the degradation of Nrf2. Further, Sch suppresses phosphorylation of P65 and its nuclear translocation, as well as the degradation of IκBα. Collectively, our findings reveal that Sch protects against OVX-induced bone loss by suppressing ROS via Nrf2. CONCLUSIONS: Our results showed the potential of anti-oxidant compound schisandrin A in the treatment of osteoporosis, highlighting Nrf2 as a novel promising target in osteoclast-related disease.