CDDO-Im ameliorates osteoarthritis and inhibits chondrocyte apoptosis in mice via enhancing Nrf2-dependent autophagy.

Osteoarthritis (OA) is the most prevalent chronic degenerative joint disease with few treatment options. The pathogenesis of OA is characterized by sustained inflammation, oxidative stress and chondrocyte apoptosis that eventually lead to cartilage degradation and joint dysfunction. In the present study, we identified a synthetic triterpenoid CDDO-Im(1-[2-cyano-3,12-dioxooleana-1,9(11)-dien-28-oyl] imidazole) as an activator of Nrf2 (nuclear factor erythroid 2-related factor 2) that displayed strong anti-OA effects. We showed that CDDO-Im (20 nM) significantly alleviated TNF-α-induced apoptosis of primary human chondrocytes and extracellular matrix degradation. In a mouse OA model incurred by DMM (destabilization of medial meniscus), administration of CDDO-Im (2.5 mg/kg, ip, every other day for 8 weeks) effectively reduced knee joint cartilage erosion and serum levels of inflammatory cytokines IL-1ß and IL-6. We revealed that CDDO-Im (20 nM) significantly enhanced autophagy activities in chondrocytes, whereas the autophagy inhibition by chloroquine (CQ, 50 µM) or 3-methyladenine (3-MA, 5 mM) abrogated the anti-apoptosis and chondroprotective effects of CDDO-Im in TNF-α-treated chondrocytes. Moreover, we confirmed that CDDO-Im (1-20 nM) dose-dependently activated Nrf2 pathway in TNF-α-treated chondrocytes, and its chondroprotective and autophagy-enhancing effects were significantly diminished when Nrf2 signaling was blocked by Nrf2 inhibitor ML385 (20 µM) or siRNA-mediated Nrf2 knockdown. Together, our results demonstrate that CDDO-Im exhibits prominent chondroprotective and anti-OA activities owing to its Nrf2 activation and autophagy-enhancing properties, which might provide new insights into the strategies of OA clinical prevention and treatment.

Knockdown of Ggps1 in chondrocyte expedites fracture healing by accelerating the progression of endochondral ossification in mice.

Bone fracture healing is achieved through the proliferation and differentiation of stem cells, while bone marrow stem cells (BMSCs) contribute to endochondral ossification. During fracture healing, mesenchymal progenitor cells first form a cartilaginous blastema that becomes vascularized to recruit precursor cells of osteoblasts through the bone morphogenetic protein 2 (Bmp2)/Smad-dependent Runx2 pathway. Statins deplete geranylgeranyl diphosphate (GGPP), which participates in the regulation of BMSCs differentiation, through the inhibition of 3-hydroxy-3-methylglutaryl (HMG)-CoA reductase, leading to impaired protein geranylgeranylation, which strongly impacts the bone synthesis induced by Bmp2. Accordingly, we would like to investigate the role of geranylgeranyl diphosphate synthase 1 (Ggps1) in bone fracture via endochondral ossification in mice. We used a Cre-loxP system, namely the tamoxifen-inducible Collagen 2-CreERT2 Ggps1 fl/fl, to eliminate specifically the Ggps1 activity in chondrocytes of 8-10-week-old mice. We found that the endochondral bone formation, calcification and vasculogenesis of the bony callus were accelerated in fractures in Ggps1-/-mice. Together, the results of this study confirm that the specific deletion of Ggps1, using the Collagen 2-CreERT2 mice, will accelerate the fracture healing process by activating the Bmp2/Smad-dependent Runx2 pathway. In addition, we managed to improve the fracture healing process by inhibiting the Ggps1 activity and its related products with statin drugs.

Fenofibrate decreases the bone quality by down regulating Runx2 in high-fat-diet induced Type 2 diabetes mellitus mouse model.

BACKGROUND: This study is to investigate the effect of fenofibrate on the bone quality of Type 2 diabetes mellitus (T2DM) mouse model. METHODS: T2DM mouse model was induced by high-fat-diet, and the mice were treated with fenofibrate (100 mg/kg) (DIO-FENO) or PBS (DIO-PBS) for 4 weeks. The bone microstructure and biomechanical properties of femora were analyzed by micro-CT and 3-Point bending test. The protein expression was detected by immunohistochemical staining and Western blot. The cell apoptosis was evaluated by TUNEL staining. The Bcl2, caspase 3, and osteoblast marker genes were detected by RT-qPCR. RESULTS: The biomechanical properties of bones from DIO-FENO group were significantly lower than those in the control and DIO-PBS groups. Besides, the trabecular number was lower than those of the other groups, though the cortical porosity was decreased compared with that of DIO-PBS group because of the increase of apoptotic cells. The expression of osteocalcin and collagen I were decreased after treatment with fenofibrate in T2DM mice. Moreover, the cell viability was decreased after treated with different concentrations of fenofibrate, and the expression of Runx2 decreased after treated with high dose of fenofibrate. CONCLUSION: Fenofibrate decreases the bone quality of T2DM mice through decreasing the expression of collagen I and osteocalcin, which may be resulted from the down regulation of Runx2 expression.

Inhibition of Wnt/ß-catenin signaling suppresses bleomycin-induced pulmonary fibrosis by attenuating the expression of TGF-ß1 and FGF-2.

Pulmonary fibrosis is a progressive lung disorder of unknown etiology, which is characterized by alterations in alveolar epithelium function, fibroblast activation, and increased extracellular matrix deposition. Recent studies have demonstrated that PF is associated with uncontrolled production of cytokines after lung injury. In the present study, we found that transforming growth factor-ß1 (TGF-ß1) and fibroblast growth factor 2 (FGF-2) were both upregulated in bleomycin-induced fibrotic lung tissue and primary murine alveolar epithelial Type II (ATII) cells treated with bleomycin. Furthermore, we discovered that TGF-ß1 could induce the differentiation of lung resident mesenchymal stem cells (LR-MSCs) into fibroblasts, which may play an essential role in PF. LR-MSCs incubated with FGF-2 showed modest alterations in the expression of α-SMA and Vimentin. Moreover, in our study, we found that Wnt/ß-catenin signaling was activated both in vitro and in vivo as a result of bleomycin treatment. Interestingly, we also found that suppression of the Wnt/ß-catenin signaling could significantly attenuate bleomycin-induced PF accompanied with decreased expression of TGF-ß1 and FGF-2 in vitro and in vivo. These results support that controlling the aberrant expression of TGF-ß1 and FGF-2 via inhibition of Wnt/ß-catenin signaling could serve as a potential therapeutic strategy for PF.

NFC-enabled potentiostat and nitrocellulose-based metal electrodes for electrochemical lateral flow assay.

Rapid detection of pathogens at the point-of-need is crucial for preventing the spread of human, animal and plant diseases which can have devastating consequences both on the lives and livelihood of billions of people. Colorimetric, lateral flow assays consisting of a nitrocellulose membrane, are the preferred format today for low-cost on-site detection of pathogens. This assay format has, however, historically suffered from poor analytical performance and is not compatible with digital technologies. In this work, we report the development of a new class of digital diagnostics platform for precision point-of-need testing. This new versatile platform consists of two important innovations: i) A wireless and batteryless, microcontroller-based, low-cost Near Field Communication (NFC)-enabled potentiostat that brings high performance electroanalytical techniques (cyclic voltammetry, chronoamperometry, square wave voltammetry) to the field. The NFC-potentiostat can be operated with a mobile app by minimally trained users; ii) A new approach for producing nitrocellulose membranes with integrated electrodes that facilitate high performance electrochemical detection at the point-of-need. We produced an integrated system housed in a 3D-printed phone case and demonstrated its use for the detection of Maize Mosaic Virus (MMV), a plant pathogen, as a proof-of-concept application.

[SP13786 Inhibits the Migration and Invasion of Lung Adenocarcinoma Cell A549 
by Supressing Stat3-EMT via CAFs Exosomes].

BACKGROUND: Fibroblast activation protein (FAP) is one of the surface markers of cancer-associated fibroblasts (CAFs) and is closely related to the malignant characterization of CAFs. SP13786 is a specific micromolecule inhibitor of FAP and this study is to investigate the effects and mechanism of SP13786 on the migration and invasion of A549 cells through regulating exosomes of CAFs. METHODS: CAFs and paracancerous fibroblasts (PTFs) were isolated and subcultured from freshly resected lung adenocarcinoma tissues and paracancerous normal tissues separately. MTT assay was used to detect the proliferation of CAFs incubated by different concentrations of SP13786; PTFs-exo, CAFs-exo and CAFs+SP13786-exo were extracted by polymer precipitation method. The A549 cells were divided into Ctrl group, PTFs group, CAFs group and SP13786 group and each group was incubated with DMEM, PTFs-exo, CAFs-exo and CAFs+SP13786-exo separately. Laser confocal microscope was used to observe the endocytoses of exosomes by A549 cells. The expression of alpha-smooth muscle actin (α-SMA) and FAP in PTFs and CAFs and the expression of E-cadherin, N-cadherin, Slug, Stat3 and P-Stat3 in A549 cells were detected by immunofluorescence, immunohistochemistry and Western blot. The migration and invasion ability of A549 cells were detected by cell scratch and transwell methods. RESULTS: α-SMA and FAP were expressed much higher in CAFs than that in PTFs which indicate that CAFs and PTFs were successfully obtained from lung adenocarcinoma and paracancerous tissues (P<0.05). MTT showed that the 50% inhibitory concentration (IC50) of SP13786 for CAFs was about 3.3 nmol/L. In addition, SP13786 can significantly decrease the expression of α-SMA and FAP in CAFs which means that targeted inhibition of FAP could reduce the malignant characteristics of CAFs (P<0.05). Laser confocal microscope found that exosomes from CAFs could be taken up by A549 cells and scratch and transwell tests showed that the endocytosed CAFs-exo could promote the migration and invasion of A549 cells (P<0.001), while FAP inhibitor SP13786 could inhibit the effects of CAFs-exo on A549 cells (P<0.05). Furthermore, Immunofluorescence and Western blot showed that CAFs-exo could promote EMT by decreasing E-cadherin expression and increasing N-cadherin, Slug expression in A549 cells while FAP inhibitor SP13786 could significantly supress CAFs-exo-induced epithelial-mesenchymal transition (EMT) of A549 cells (P<0.05). Moreover, the expression of P-Stat3 was obviously increased in A549 cells of CAFs group and significantly down-regulated in SP13786 group (P<0.05) whereas there was no significant difference in total Stat3 between CAFs and SP13786 groups (P>0.05). Finally, WP1066 (a specific inhibitor of Stat3) was used to comfirm whether SP13786 could influence EMT of A549 cells by inhibiting Stat3 phosphorylation via CAFs-Exo. The results showed that when the phosphorylation of Stat3 in CAFs group was inhibited by WP1066, SP13786 could not influence the P-Stat3 expression and EMT of A549 cells anymore (P>0.05). CONCLUSIONS: As a specific micromolecule inhibitor of FAP, SP13786 indirectly inhibits the migration and invasion of A549 cells by affecting exosomes of CAFs. The possible mechanism is to inhibit the phosphorylation of Stat3 and thus affect the EMT of A549 cells.

One pot synthesis of zwitteronic 99mTc doped ultrasmall iron oxide nanoparticles for SPECT/T1-weighted MR dual-modality tumor imaging.

In this study, we have synthesized 99mTc intrinsically labeled ultrasmall magnetic iron oxide nanoparticles with zwitterionic surface coating (99mTc-ZW-USIONPs) via one pot synthesis using sulfobetains functionalized poly (acrylic acid) as stabilizer and Na99mTcO4 and SnCl2 as additives. The commercialization of single photon emission computed tomography (SPECT)/magnetic resonance imaging (MRI) scanner made the combination use of 99mTc and iron oxide nanoparticles attracting much attention. Direct doping radioisotope into nanoparticles has the advantages of excellent radiochemical stability and no restriction on the surface functionalization. The complex Technetium chemistry made it challenging to direct dope 99mTc into IONPs, especially those ultrasmall ones without precipitation. We proved that it is possible to prepare 99mTc doped USIONPs with excellent water solubility and favorable T1 signal by controlling the radioactivity and reducing agent amount. With no need of chelator, the zwitterionic surface resists the protein corona formation, resulting in a reduced RES uptake and higher tumor contrast. The 99mTc-ZW-USIONPs demonstrated excellent performance of tumor SPECT and T1-weighted MR imaging capability in 4T1 tumor bearing mice. Together with their ease of preparation and superior biocompatibility, we believe these 99mTc-ZW-USIONPs represent a type of promising dual contrast agent for SPECT/T1 MRI.

Three-dimensional finite-element analysis of aggravating medial meniscus tears on knee osteoarthritis.

BACKGROUND: The biomechanical change during the medial meniscus damage in the process of knee osteoarthritis has not been explored. The purpose of this study was to determine the effect of aggravating medial meniscus degenerative tear on the progress of knee osteoarthritis through the finite-element simulation method. METHODS: The three-dimensional digital model of a total-knee joint was obtained using a combination of magnetic resonance imaging and computed tomography images. Four types of medial meniscus tears were created to represent the aggravating degenerative meniscus lesions. Meniscectomy of each meniscal tear was also utilized in the simulation. The compression and shear stress of bony tissue, cartilage, and meniscus were evaluated, and meniscus extrusion of the healthy knee, postinjured knee, and postmeniscectomy knee were investigated under the posture of balanced standing. RESULTS: Based on the results of finite-element simulation, the peak shear principal stress, peak compression principal stress, and meniscus extrusion increased gradually as the meniscus tears' region enlarged progressively (from 7.333 MPa to 15.14 MPa on medial femur and from 6 MPa to 20.94 MPa on medial tibia). The higher stress and larger meniscus extrusion displacement in all tests were observed in the flap and complex tears. The oblique tears also had a biomechanical variation of stress and meniscus extrusion in the knee joint, but their level was milder. Both the peak value of the stress and meniscus displacement increased after the meniscectomy. CONCLUSION: In contrast to the damaged hemijoint, the stress applied on the healthy lateral hemijoint increased. The change of biomechanics was more obvious with the aggravation of meniscus injury. The advanced degenerative damage resulted in increasing stress that was more likely to cause symptomatic clinical manifestation in the knee joint and accelerate the progress of osteoarthritis. Moreover, we found that the meniscus injury caused higher stress concentration on the contralateral side of the joint. We also discovered that the meniscectomy can lead to more serious biomechanical changes, and although this technique can relieve pain over a period of time, it increased the risk of osteoarthritis (OA) occurrence. THE TRANSLATIONAL POTENTIAL OF THIS ARTICLE: It is clear that the meniscal lesions can cause osteoarthritic knee, but the biomechanical change during the meniscus damage period has not been explored. We have evaluated the variation of stress during the aggravating medial degenerative meniscus tears and the relationship in the process of knee OA through finite-element simulation. This study does favour to obtain a better understanding on the symptoms and pathological changes of OA. It also may provide some potential directions for the prophylaxis and treatment of OA.

Early osteointegration evaluation of porous Ti6Al4V scaffolds designed based on triply periodic minimal surface models.

BACKGROUND: The graded porous structures were designed using triply periodic minimal surfaces models to mimic the biomechanical properties of bone. The mechanical properties and bone formation ability were evaluated to explore the feasibility of the design method in bone tissue engineering. METHODS: The scaffolds were designed using a P-surface with different pore sizes. All materials were fabricated using 3D printing technology and the mechanical properties were tested by an electronic universal testing device. The biomechanical properties were then analyzed by finite element method, while the ontogenesis of the material in vivo was examined by implanting the scaffolds for five weeks in pigs. RESULTS: According to the obtained results, the pore size ranged between 100 µm to about 700 µm and porosity were around 49.54%. The graded porous architectures can decrease the stiffness of implants and reduce the stress shielding effect. In addition, these porous structures can stimulate bone ingrowth and achieve a stable interface between implants and surrounding bone tissues after 5 weeks' implantation. The micro-CT results also demonstrated the obviously bone formation around all the porous structures. CONCLUSION: To sum up, the triply periodic minimal surfaces based graded porous structure is effective in decreasing the stress shielding effect, promoting early osteogenesis and osteointegration. This is the first research to explore the effect of this kind of porous structures on bone formation in vivo where the obtained results supported the previous theoretical research on the application potential in bone tissue engineering. THE TRANSLATIONAL POTENTIAL OF THIS ARTICLE: Porous architecture designed using triply periodic minimal surface models can achieve gradually changed pore size and appropriate porosity for bone regeneration. This kind of structure can mimic the Young's modulus of natural bone tissue, improve the stress transmission capability and dismiss the stress shielding effect. It also can stimulate the early bone integration in vivo and enhance the binding force between bone and implants, which may bring a new design method for orthopaedic implants and their surface structure.

Effect of degenerative and radial tears of the meniscus and resultant meniscectomy on the knee joint: a finite element analysis.

OBJECTIVE: The objective of this study is to investigate the biomechanics on the knee components caused by degenerative and radial meniscal tears and resultant meniscectomy. METHODS: A detailed finite element model of the knee joint with bones, cartilages, menisci and main ligaments was constructed from a combination of computed tomography and magnetic resonance images. Degenerative and radial tears of both menisci and resultant medial meniscectomy were used and two different kinds of simulations, the vertical and the anterior load, mimicking the static stance and slight flexion simulations, were applied on the model. The compressive and shear stress and meniscus extrusion were evaluated and compared. RESULTS: Generally, both degenerative and radial tears lead to increased peak compressive and shear stress of both cartilages and menisci and large meniscus extrusion, and the medial meniscal tear induced larger value of stress and extrusion than the lateral meniscal tear. The peak stress and meniscus extrusion further elevated after the medial meniscus meniscectomy. Distribution of stress was shifted from the intact hemi joint to the injured hemi joint with either medial or lateral meniscal tear. CONCLUSION: Our finite element model provides a realistic three-dimensional knee model to investigate the effects of degenerative and radial meniscal tears and resultant meniscectomy on the stress distribution of the knee. The stress was increased in meniscal tears and increased significantly when meniscectomy was performed. Increased meniscus extrusion may explain the mechanism for higher stress on the components of the knee. THE TRANSLATIONAL POTENTIAL OF THIS ARTICLE: Meniscal tears are the most common damage associated to the menisci, and meniscectomy is often performed to relieve the pain and instability of the knee. The results of our study indicated increased stress on cartilages and menisci, which may lead to early onset of osteoarthritis. This may guide surgeons to preserve more of the meniscus when performing meniscectomy.

Stability evaluation of anterior external fixation in patient with unstable pelvic ring fracture: a finite element analysis.

BACKGROUND: The pelvic ring fractures (PRF) are commonly induced by the high-energy impact and will lead to unstable and sever injures. This study is aimed to explore the stability of anterior external fixation in treating pelvis fracture and evaluate the possibility for these kinds of patients to reduce bedridden time. METHODS: A patient with Tile B3 pelvis fracture was chosen in the research and the corresponding digital model was reconstructed according to the CT images and 3D scanning. Four angles of pelvis under vertical compression were employed in the finite element (FE) analyses. The stress distribution and micro-motion displacement were calculated to validate the instability of pelvis. RESULTS: The stress applied on the pelvis was ranged from 4.296 to 8.364 MPa in all postures. The stress applied on pins was less than 7.011 MPa during reclining, and reached 28.29 MPa when standing. The micro-motion displacement in reclining posture was ranged from 0.005 to 0.087 mm. The value increased to more than 1mm in standing posture. CONCLUSIONS: It was safety for patients with pelvis fracture to sit vertical or recline on the bed during nursing or having treatment, but standing or walking will generate inappropriate micro-motion. The existence of external fixation can reduce the possibility of complications caused by long-term bedridden.

The biomechanical changes of load distribution with longitudinal tears of meniscal horns on knee joint: a finite element analysis.

BACKGROUND: Meniscal horns are important structures of meniscus, and longitudinal tears of these places could significantly change the load distribution among the knee joint. Few studies concerned the stress concentrated on bones, which may induce the osteonecrosis of subchondral bone. The goal of this study was to construct a finite element (FE) model with high fidelity of the knee joint and evaluate the biomechanical changes of load distribution of components after longitudinal tears of the horns of meniscus. METHODS: Computed tomography and magnetic resonance images were used to develop the FE model, and two different kinds of simulations, the vertical and the anterior load, mimicking the static stance and slight flexion simulations, were applied after longitudinal tears of the horns of meniscus. RESULTS: Significantly elevated peak compressive and shear stress was observed on the menisci, cartilages, and subchondral bones, and enlarged meniscus extrusion was noticed. Between all the four types of longitudinal tears investigated in this study, longitudinal tears at the posterior horn of the medial meniscus were found to be the most significant. CONCLUSIONS: These findings showed that longitudinal tears of the meniscal horns lead to increased magnitude and changed distribution of stress and indicated the important role of posterior horn of medial meniscus. This may contribute to the mechanism between meniscal tears and spontaneous subchondral bone osteonecrosis.

Biomechanical analysis of the effect of medial meniscus degenerative and traumatic lesions on the knee joint.

The purpose of this study was to determine the effect of the degenerative medial meniscus and traumatic lesions on the biomechanical behavior of the knee. An elaborate three-dimensional (3D) finite element model of the total knee joint containing bones, articular cartilages, main ligaments, and menisci was developed from a combination of magnetic resonance images and computed tomography. Three types of meniscus tears were employed to represent the degenerative and traumatic lesions. The stress and meniscus extrusion of healthy and injured knees were investigated under the posture of static stance. The traumatic longitudinal tear demonstrated the highest stress and the largest meniscus extrusion displacement. The degenerative horizontal and peripheral tears also showed an irregular biomechanical balance in the knee joint. Despite the damaged hemijoint, the stress on the healthy lateral hemijoint was increased. Although the biomechanics was deteriorated in all meniscus tear models, the variation degree was diverse. The transfixion damage could potentially cause future injury in the knee joint and accelerate the progress of osteoarthritis. Moreover, the meniscus injury may cause high-stress concentration on the contralateral side of the joint. The current results revealed the cause of different clinical manifestation after meniscus tears and the risk of knee osteoarthritis through biomechanical aspects.

Polydopamine coating promotes early osteogenesis in 3D printing porous Ti6Al4V scaffolds.

BACKGROUND: Titanium implants are widely used in orthopedic and dental for more than 30 years. Its stable physicochemical properties and mechanical strength are indeed appropriate for implantation. However, the Bioinertia oxidized layer and higher elastic modulus often lead to the early implantation failure. METHODS: In this study, we proposed a simple design of porous structure to minimize the disparity between scaffold and natural bone tissue, and introduced a one-step reaction to form a polydopamine (PDA) layer on the surface of titanium for the purpose of improving osteogenesis as well. The porous scaffolds with pore size of 400 µm and porosity of 44.66% were made by additive manufacturing. The cell behavior was tested by seeding MC3T3-E1 cells on Ti6Al4V films for 15 days. The biomechanical properties were then analyzed by finite element (FE) method and the in vivo osteogenesis effect was accordingly evaluated by implanting the scaffolds for 5 weeks in rabbits. RESULTS: According to the achieved results, it was revealed that the immersion for 40 min with dopamine could significantly improve the cell adhesion. The proposed method for design of porous structure can avoid the stress shielding effect and bone growth inside the PDA coating scaffolds, which were observed at the early stage of bone healing process. CONCLUSIONS: It can be concluded that the proposed PDA coating method is effective in promoting early osteogenesis, as well as being easy to operate, and can be helpful in the future clinical application of titanium implants.

Inhibition of pyroptosis attenuates Staphylococcus aureus-induced bone injury in traumatic osteomyelitis.

BACKGROUND: Osteomyelitis is a severe bone infection and typically leads to progressive bone resorption, destruction and dysfunction. Pyroptosis is a form of programmed cell death involved in various infectious diseases. However, the identification of pyroptosis and the role it plays in osteomyelitis remains to be clarified. In this study, we investigated the expression of pyroptosis-associated proteins in osteomyelitis and the effects of inhibiting pyroptosis on S. aureus-induced osteomyelitis both in vitro and in vivo. METHODS: The expression of pyroptosis-associated protein-NLRP3 (NLR Family Pyrin Domain Containing 3), Caspase1 and GSDMD (GasderminD) were examined in murine and human infectious bone fragments by western blot. Bone destruction was evaluated by microcomputed tomography (µCT). The concentration of inflammatory factors was tested by Enzyme linked Immunosorbent Assay (ELISA). The expression of pyroptosis-associated gene was detected by real-time quantitative polymerase chain reaction (RT-qPCR). RESULTS: The expression of pyroptosis-associated proteins in infectious bone fragments from patients with osteomyelitis was significantly higher than uninfected bone. Additionally, in S. aureus-induced murine osteomyelitis model, higher expression of pyroptosis-associated proteins was noticed. Furthermore, the inhibitors of pyroptosis-associated proteins alleviated S. aureus-induced pyroptosis both in vivo and in vitro. More importantly, the inhibition of pyroptosis restored the bone formative property, attenuated the aberrant activation of osteoclast in vitro and reversed bone injury in vivo. CONCLUSIONS: Our study identified pyroptosis as a key pathway in osteomyelitis and elaborated that the inhibition of pyroptosis could attenuate S. aureus-induced bone destruction in osteomyelitis, providing a potential treatment target to osteomyelitis.

The effectiveness of allogeneic mesenchymal stem cells therapy for knee osteoarthritis in pigs.

BACKGROUND: Intraarticular injection of the mesenchymal stem cells (MSCs) has shown to be successful for treating osteoarthritis (OA). Nevertheless, many studies have been focusing on autologous MSCs. The following study investigates the safety and effectiveness of intraarticular injection of allogenic MSCs in a pig OA model. METHODS: Superparamagnetic iron oxide (SPIO) nanoparticles were labelled with bone marrow-derived mesenchymal stem cells (BM-MSCs) to allow cells tracking using magnetic resonance imaging (MRI). A pig OA model was established by bilateral medial meniscectomy. Next, SPIO-BM-MSCs were injected into the right knee, while the left knee was left untreated. MRI and radiography were used to assess the degree of OA and to evaluate the effectiveness of allogenic MSCs. Hematoxylin and eosin (H&E), safranin-o fast green staining, toluidine blue, and immunohistochemical staining were used to evaluate the therapeutic effect of the injections. RESULTS: At concentration of ≤20 µg/mL, SPIO caused no toxicity to BM-MSCs. Four weeks after surgery, OA changes were observed on MRI scan. The SPIO labeled BM-MSCs were found moving towards the impaired part of the cartilage 8 to 24 h after injections. In addition, no significant differences between the right side (therapeutic side) and the left side (untreated side) were observed following histological and immunohistochemistry analysis. CONCLUSIONS: The suitable concentration of SPIO for labelling BMSCs was 20 µg/mL, while the allogenic MSCs could move towards and accumulate around the impaired cartilage. No significant difference was found between treatment and control group.

Research progress of the relationship between pyroptosis and disease.

Pyroptosis, characterized by proinflammation, has been defined as a new type of programmed cell death in recent years. Inflammasomes are activated by the corresponding pathogen-associated molecular patterns (PAMPS) or damage-associated molecular patterns (DAMPS), followed up by the cleavage of pro-interleukin-1ß (pro-IL-1ß), pro-interleukin-18 (pro-IL-18) and gasdermin D. The N-terminal fragment of gasdermin D gives rise to the destruction of cell membrane, leading to cell rupture as well as the efflux of proinflammatory cytokines. Recent studies have shown that pyroptosis is associated with a variety of diseases due to its proinflammation effect and the dysfunction of related cells. The relationship between pyroptosis and associated diseases is described in this review.

Zhuangguguanjie formulation protects articular cartilage from degeneration in joint instability-induced murine knee osteoarthritis.

Zhuangguguanjie formulation (ZG) can provide noticeable relief from joint pain in patients suffering from knee osteoarthritis (OA). However, the underlying mechanism has not been fully described. Male C57BL/6 mice were administered either ZG or normal saline (NS) following surgical destabilization of the medial meniscus (DMM). At weeks 4, 6 and 8 (post-surgery), knee joints were harvested and assessed with Safranin-O staining. Blood serum was collected and tested. In vitro analysis was carried out to evaluate the effects of ZG on the expression of the OA-related genes. DMM mice indicated reduced cartilage destruction and lower blood serum biomarkers of OA (COMP1 and CTX-1) following ZG treatment. Moreover, the femoral condyle and tibial plateau histological scores were significantly reduced following ZG treatment of the DMM mice. ZG could markedly downregulate the expression of OA-related genes namely, ADAMTS5, MMP3 and MMP13, while it simultaneously upregulated collagen II as demonstrated by in vitro assays. Moreover, chondrocyte apoptosis was significantly decreased following ZG treatment. These results may be caused by the up-regulation of p-AKT expression levels, since the anti-apoptotic effects of ZG can be blocked by treatment with an AKT inhibitor. ZG is capable of preventing and/or reducing the progression of OA by inhibiting chondrocyte apoptosis via the p-AKT/Caspase 3 pathway.

Evaluation of a polyvinyl alcohol-alginate based hydrogel for precise 3D bioprinting.

In this study, we designed a polyvinyl alcohol (PVA)-alginate based hydrogel and evaluated its cytocompatibility and printability. The samples were fabricated by 3D printing using a freeze-thaw process. The scanning electron microscope, material testing machine, rheometer, and cell counting kit-8 assay were used to examine the morphology, mechanical properties, rheological properties, and cytocompatiblity of the scaffolds, respectively. The mechanical strength, cytocompatiblity, crosslinking time, and printability were remarkably improved with the use of PVA. To sum up, our data suggest that hybrid bio-ink is more appropriate for precise 3D bioprinting due to its rapid prototyping capability and better cytocompatibility. © 2018 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 106A: 2944-2954, 2018.

Egr1 deficiency disrupts dynamic equilibrium of chondrocyte extracellular matrix through PPARγ/RUNX2 signaling pathways.

BACKGROUND: This study is to investigate the effect of Egr1 on the mineralization and accumulation of chondrocyte extracellular matrix. METHODS: The femoral heads of patients of various heights were collected. Egr1 knockout mice were used. Their limb lengtha nd body weight were assessed. The bone characteristics were detected by micro-CT scan and histological staining. Immature murine articular chondrocytes (iMACs) were isolated. Gross morphology was observed by histological staining. Relevant mRNA and protein expression were detected by qRT-PCR and Western blot, respectively. the related proteins were observed by immunohistochemical staining and immunofluorescence assay. Chromatin immunoprecipitation and reporter gene assay were also used. TUNEL was used to detect apoptosis. RESULTS: It was found that shorter patients had reduced Egr1 expression levels in the hypertrophic cartilage zone of the femoral head. In addition, Egr1 knockout mice exhibited reduced body size. Micro-CT analysis showed that these mice also had reduced bone volume. Safranin-O staining showed that the extracellular matrix of these mice exhibited a relatively limited degree of mineralization, and TUNEL staining showed reduced cell apoptosis levels. After transfecting the iMACs with dominant-negative Egr1 adenoviruses to inhibit Egr1, the enzymes of Adamst4, Adamst5, Mmp3 and Mmp13 were significantly upregulated. ChIP and luciferase assays revealed that Egr1 might regulate the chondrocyte extracellular matrix by the PPARγ/RUNX2 signaling pathways. CONCLUSION: Egr1 has an important regulatory effect on the dynamic equilibrium of the chondrocyte extracellular matrix, which may be achieved through the PPARγ/RUNX2 signaling pathways.