Sappanone a alleviates osteoarthritis progression by inhibiting chondrocyte ferroptosis via activating the SIRT1/Nrf2 signaling pathway.

Osteoarthritis (OA) is a common degenerative joint disease that cause pain and disability in adults. Chondrocyte ferroptosis is found to be involved in OA progression. Sappanone A has been found as an anti-inflammatory and antioxidative agent in several diseases. This study aims to investigate the effects of sappanone A on OA progression and chondrocyte ferroptosis. IL-1ß-induced chondrocytes and destabilization of the medial meniscus (DMM)-induced rats were respectively used as the OA model in vitro and in vivo. The effects of sappanone A on inflammation, extracellular matrix (ECM) metabolism, and ferroptosis were determined. Our results showed that in IL-1ß-induced chondrocytes, sappanone A suppressed the production of NO, PGE2, TNF-α, IL-6, iNOS, and COX2. Sappanone A also inhibited the expression of MMP3, MMP13, and ADAMTS5, while increasing collagen II expression. Moreover, sappanone A alleviated cytotoxicity and decreased the levels of intracellular ROS, lipid ROS, MDA, and iron, while increasing GSH levels. Additionally, sappanone A increased the protein expression of SLC7A11 and GPX4. Administration of ferroptosis activator reversed the inhibitory effects of sappanone A on IL-1ß-induced inflammation and ECM degradation. More importantly, Sappanone A activated the Nrf2 signaling by targeting SIRT1. The inhibition of sappanone A on ferroptosis was greatly eliminated due to the addition of SIRT1 inhibitor. Furthermore, intra-articular injection of sappanone A mitigated cartilage destruction and ferroptosis in DMM-induced OA rats. In conclusion, sappanone A protects against inflammation and ECM degradation in OA via decreasing chondrocyte ferroptosis by activating the SIRT1/Nrf2 signaling. These findings deepen our understanding of chondrocyte ferroptosis in OA and highlight the therapeutic potential of sappanone A for OA.

Tumour necrosis factor α regulates the miR-27a-3p-Sfrp1 axis in a mouse model of osteoporosis.

Osteoporosis is a metabolic bone disease that involves gradual loss of bone density and mass, thus resulting in increased fragility and risk of fracture. Inflammatory cytokines, such as tumour necrosis factor α (TNF-α), inhibit osteogenic differentiation of bone marrow mesenchymal stem cells (BMSCs), and several microRNAs are implicated in osteoporosis development. This study aimed to explore the correlation between TNF-α treatment and miR-27a-3p expression in BMSC osteogenesis and further understand their roles in osteoporosis. An osteoporosis animal model was established using ovariectomized (OVX) mice. Compared with Sham mice, the OVX mice had a significantly elevated level of serum TNF-α and decreased level of bone miR-27a-3p, and in vitro TNF-α treatment inhibited miR-27a-3p expression in BMSCs. In addition, miR-27a-3p promoted osteogenic differentiation of mouse BMSCs in vitro, as evidenced by alkaline phosphatase staining and Alizarin Red-S staining, as well as enhanced expression of the osteogenic markers Runx2 and Osterix. Subsequent bioinformatics analysis combined with experimental validation identified secreted frizzled-related protein 1 (Sfrp1) as a downstream target of miR-27a-3p. Sfrp1 overexpression significantly inhibited the osteogenic differentiation of BMSCs in vitro and additional TNF-α treatment augmented this inhibition. Moreover, Sfrp1 overexpression abrogated the promotive effect of miR-27a-3p on the osteogenic differentiation of BMSCs. Furthermore, the miR-27a-3p-Sfrp1 axis was found to exert its regulatory function in BMSC osteogenic differentiation via regulating Wnt3a-ß-catenin signalling. In summary, this study revealed that TNF-α regulated a novel miR-27a-3p-Sfrp1 axis in osteogenic differentiation of BMSCs. The data provide new insights into the development of novel therapeutic strategies for osteoporosis.

PUD@HA/PEEK Scaffold Induces Subchondral Bone Regeneration to Repair Osteochondral Defect in Rabbits.

Osteochondral defects (OCDs) pose a significant challenge in clinical practice, and recent advancements in their repair indicate that satisfying subchondral bone repair may be critical for this. Herein, a series of hydroxyapatite/poly(ether ether ketone) (HA/PEEK) scaffolds were fabricated with varying mass percentages (0, 20, 30, and 40%) to induce subchondral bone regeneration. Subsequently, an optimal scaffold with 40% HA/PEEK was selected to establish osteochondral scaffolds with poly(ether urethane) urea-Danshensu (PUD) for repairing the OCD. The material characteristics of HA/PEEK and PUD were investigated using scanning electron microscopy, tensile, swelling, and fatigue tests, and cytological experiments. The effects of serial HA/PEEK scaffolds on subchondral bone repair were then assessed by using microcomputed tomography, hard tissue slicing, and histological staining. Furthermore, the optimal 40% HA/PEEK scaffold was used to develop osteochondral scaffolds with PUD to observe the effect on the OCD repair. HA/PEEK materials exhibited an even HA distribution in PEEK. However, when composited with HA, PEEK exhibited inferior mechanical strength. 40%HA/PEEK scaffolds showed an optimum effect on in vivo subchondral bone repair. Cartilage regeneration on 40%HA/PEEK scaffolds was pronounced. After PUD was introduced onto the HA/PEEK, the PUD@40%HA/PEEK scaffold produced the expected effect on the repair of the OCD in rabbits. Therefore, achieving satisfactory subchondral bone repair can benefit surficial cartilage repair. The PUD@40%HA/PEEK scaffold could induce subchondral bone regeneration to repair the OCD in rabbits and could provide a novel approach for the repair of the OCD in clinical practice.

Ultraviolet Light Responsive N-Nitroso Polymers for Antibacterial Nitric Oxide Delivery.

This study investigates the incorporation of active secondary amine moieties into the polymer backbone by co-polymerizing 2,4,6-tris(chloromethyl)-mesitylene with three diamines, namely 1,4-diaminobutane, m-phenylenediamine, and p-phenylenediamine. This process results in the stabilization of the amine moieties and the subsequently introduced nitroso groups. Charging bioactive nitric oxide (NO) into the polymers is accomplished by converting the amine moieties into N-nitroso groups. The ability of the polymers to store and release NO depends on their structures, particularly the amount of incorporated active secondary amines. With grafting photosensitive N-nitroso groups into the polymers, the derived NO@polymers exhibit photoresponsivity. NO release is completely regulated by adjusting UV light irradiation. These resulting polymeric NO donors demonstrate remarkable bactericidal and bacteriostatic activity, effectively eradicating E. coli bacteria and inhibiting their growth. The findings from this study hold promising implications for combining NO delivery with phototherapy in various medical applications.

Direct detection and identification of periprosthetic joint infection pathogens by metagenomic next-generation sequencing.

This study assessed the application of metagenomic next-generation sequencing in pathogen detection of periprosthetic joint infections. A total of 95 cases who previously had undergone hip and knee replacement undergoing revision from January 2018 to January 2021 were included in this study. Specimens of synovial fluid and deep-tissue were collected for culture and metagenomic next-generation sequencing, and patients were retrospectively categorized as infected or aseptic using the Musculoskeletal Infection Society criteria after revision surgery. The sensitivity, specificity, positive and negative predictive values were compared. A total of 36 cases had positive culture results and 59 cases had positive metagenomic next-generation sequencing results. Culture was positive in 34 infected cases (58.6%) and 2 aseptic cases (5.4%). Metagenomic next-generation sequencing was positive in 55 infected cases (94.8%) and 4 aseptic cases (10.8%). Five cases diagnosed with infection had other potential pathogens detected by metagenomic next-generation sequencing. Among the 24 culture-negative periprosthetic joint infections, metagenomic next-generation sequencing was able to identify potential pathogens in 21 cases (87.5%). From sampling to reporting, the average time needed for culture was 5.2 (95% CI 3.1-7.3) days, while that for metagenomic next-generation sequencing was 1.3 (95% CI 0.9-1.7) days. Metagenomic next-generation sequencing is more advantageous in pathogen detection of periprosthetic joint infection after total joint replacement, especially in patients with multiple infections or negative culture results.

Carbon and Iron Uptake by Phytoplankton in the Amundsen Sea, Antarctica.

Freshwater components in the Southern Ocean, whether sea ice meltwater or meteoric water, influence the growth of phytoplankton by affecting water stability and supplying dissolved iron (DFe). In addition, melting sea ice stimulates phytoplankton blooms by providing ice algae. In this study, sea ice meltwater and meteoric water in the Amundsen Sea (AS) were differentiated by their stable oxygen isotopic compositions (δ18O), while the phytoplankton carbon fixation rate (CFR) and iron uptake rate (FeUR) values were determined using the 14C and 55Fe tracer assays, respectively. Our results showed that FeUR exhibits a significant positive response only to sea ice meltwater, suggesting that DFe and algae provided by sea ice melting may be the main cause. In addition, the CFR had a slightly positive response to the freshwater input and a stronger correlation with the phytoplankton biomass, suggesting that the freshwater input may have enhanced the CFR through the algae released from sea ice melting. The FeUR normalized to the phytoplankton biomass was significantly positively correlated with the mixed layer depth, suggesting that water stability regulates the phytoplankton growth and the resulting Fe demand. A higher Fe demand per unit of carbon fixation during sea ice formation leads to a higher Fe/C ratio in phytoplankton. Although no significant correlations were observed between the FeUR, CFR, and meteoric water, meteoric water may have an effect on larger phytoplankton sensitive to Fe deficiencies. The results of culture experiments with DFe addition showed that the added Fe significantly enhanced the Fe uptake, carbon fixation, and Fe/C ratio of the cells, especially for micro-phytoplankton. The more pronounced response of micro-phytoplankton means that the meteoric water input may affect the efficiency of carbon export. Our study provides the first measurements of phytoplankton Fe quotas in the AS in austral late summer and early autumn, providing insights into how meteoric water and sea ice meltwater affect seasonal changes in Antarctic ecosystems.

Implementation of Photosensitive, Injectable, Interpenetrating, and Kartogenin-Modified GELMA/PEDGA Biomimetic Scaffolds to Restore Cartilage Integrity in a Full-Thickness Osteochondral Defect Model.

Cartilage defects caused by mechanical tear and wear are challenging clinical problems. Articular cartilage has unique load-bearing properties and limited self-repair ability. The current treatment methods, such as microfractures and autogenous cartilage transplantation to repair full-thickness cartilage defects, have apparent limitations. Tissue engineering technology has the potential to repair cartilage defects and directs current research development. To enhance the regenerative capacities of cartilage in weight-bearing areas, we attempted to develop a biomimetic scaffold loaded with a chondroprotective factor that can recreate structure, restore mechanical properties, and facilitate anabolic metabolism in larger joint defects. For enhanced spatial control over both bone and cartilage layers, it is envisioned that biomaterials that meet the needs of both tissue components are required for successful osteochondral repair. We used gelatin methacrylate (GELMA) and polyethylene glycol diacrylate (PEGDA) light-cured dual-network cross-linking modes that can significantly increase the mechanical properties of scaffolds and are capable of restoring function and prolonging the degradation time. Once the hydrogel complex was injected into the osteochondral defect, in situ UV light curing was applied to seamlessly connect the defect repair tissue with the surrounding normal cartilage tissue. The small molecule active substance kartogenin (KGN) can promote cartilage repair. We encapsulated KGN in biomimetic scaffolds so that, as the scaffold degrades, scaffold-loaded KGN was slowly released to induce endogenous mesenchymal stem cells to home and differentiate into chondrocytes to repair defective cartilage tissue. Our experiments have proven that, compared with the control group, GELMA/PEGDA + KGN repaired cartilage defects and restored cartilage to hyaline cartilage. Our study suggests that implementing photosensitive, injectable, interpenetrating, and kartogenin-modified GELMA/PEDGA biomimetic scaffolds may be a novel approach to restore cartilage integrity in full-thickness osteochondral defects.

Sappanone A Alleviated IL-1ß-Induced Inflammation in OA Chondrocytes through Modulating the NF-κB and Nrf2/HO-1 Pathways.

Objective: This study was to examine the anti-inflammatory effect of sappanone A on interleukin- (IL-) 1ß-stimulated osteoarthritis (OA) chondrocytes. Methods: Chondrocytes were pretreated with sappanone A for 2 h before subsequent IL-1ß stimulation. The mRNA expression levels of iNOs, COX-2, aggrecan, and collagen-II were measured with qRT-PCR. The levels of TNF-α, IL-6, IL-8, MMP-3, and MMP-13 were determined by ELISA. The protein levels of iNOs, COX-2, ADAMTS-4, ADAMTS-5, aggrecan, collagen-II, p-p65, p65, IκBα, Nrf2, and HO-1 were assessed by Western blot. Results: Sappanone A inhibited the IL-1ß-stimulated production of NO, PGE2, iNOS, COX-2, TNF-α, IL-6, and IL-8 in OA chondrocytes. In addition, sappanone A suppressed the expression of MMP-3, MMP-13, ADAMTS-4, and ADAMTS-5 in IL-1ß-stimulated OA chondrocytes. The degradation of ECM components was reversed by sappanone A. Sappanone A prevented NF-κB activation while enhanced Nrf2/HO-1 activation in IL-1ß-treated chondrocytes. Conclusion: Sappanone A may be a potent therapeutic agent for OA.

Dual isotopes of nitrite in the Amundsen Sea in summer.

Nitrite (NO2-) is a key intermediate in the nitrogen (N) cycle, and its transformation is accomplished by microbial communities. However, due to few studies on the nitrite cycle, a clear assessment of the contribution to the marine biogeochemical cycle is missing. Here, we present data on nitrogen and oxygen isotopic composition of NO2- in the Amundsen Sea in summer, and explore the biogeochemical processes that influence the NO2- cycle. Extremely low δ15NNO2 and abnormally high δ18ONO2 were found in the upper waters of the Amundsen Sea, with δ15NNO2 as low as -58.4 ‰ and δ18ONO2 as high as 44.4 ‰. Enzymatic isotopic exchange reactions between nitrate and nitrite have been proposed to be responsible for these isotopic anomalies. The mirror-symmetrical variation between δ15NNO2 and δ18ONO2 suggests that the isotopic fractionation effects of nitrogen and oxygen are opposite in isotope exchange reactions. Dual isotopes of nitrite indicate that ammonia oxidation is the main source of nitrite, thus nitrification plays an important role in the formation of primary nitrite maximum in the upper Amundsen Sea. The nitrogen and oxygen isotopic compositions of nitrite provide support for clarifying multiple processes of marine nitrogen cycle.

The biological uptake of dissolved iron in the changing Daya Bay, South China Sea: Effect of pH and DO.

The oceanic acidification and coastal hypoxia have potential to enhance biological uptake of dissolved iron (Fe) by phytoplankton. In this study, the Fe uptake rate (FeUR) in Daya Bay was significantly negatively correlated with pH and dissolved oxygen (DO) (r = -0.81 and -0.73, respectively, p < 0.001). In addition, binary regression (FeUR = -1.45 × pH - 0.10 × DO + 13.64) also indicated that both pH and DO played key roles in FeUR variations. As pH and DO decreased, Fe uptake by phytoplankton was promoted, and the contribution of nano-phytoplankton to Fe uptake increased significantly, while that of pico-FeUR decreased. These will result in the phytoplankton community to be miniaturized and Fe requirement of phytoplankton goes higher, thereby leading changes of phytoplankton composition and coastal ecosystem. This study helps to understand how Fe could affect the coastal ecosystem under the increasing anthropogenic influences.

Sonic Hedgehog Induces Mesenchymal Stromal Cell Senescence-Associated Secretory Phenotype and Chondrocyte Apoptosis in Human Osteoarthritic Cartilage.

Hedgehog (HH) signaling plays a critical role in osteoarthritis (OA) pathogenesis, but the molecular mechanism remains to be elucidated. We show here that Sonic Hedgehog (SHH) gene expression is initiated in human normal cartilage stromal cells (NCSC) and increased in OA cartilage mesenchymal stromal cells (OA-MSCs) during aging. Manifesting a reciprocal cellular distribution pattern, the SHH receptors PTCH1 and SMO and transcription factors GLI2 and GLI3 are expressed by chondrocytes (OAC) in OA cartilage. SHH autocrine treatment of osteoarthritis MSC stimulates proliferation, chondrogenesis, hypertrophy, and replicative senescence with elevated SASP gene expression including IL1B, IL6, CXCL1, and CXCL8. SHH paracrine treatment of OAC suppresses COL2A1, stimulates MMP13, and induces chondrocyte apoptosis. The OA-MSC conditioned medium recapitulates the stimulatory effects of SHH on OAC catabolism and apoptosis. SHH knock-down in OA-MSC not only inhibits catabolic and senescence marker expression in OA-MSC, but also abolishes the effect of the OA-MSC conditioned medium on OAC catabolism and apoptosis. We propose that SHH is a key mediator between OA-MSC and OA chondrocytes interaction in human OA cartilage via two mechanisms: (1) SHH mediates MSC growth and aging by activating not only its proliferation and chondrogenesis, but also low-grade inflammation and replicative senescence, and (2) SHH mediates OA-MSC-induced OAC catabolism and apoptosis by creating a pro-inflammatory microenvironment favoring tissue degeneration during OA pathogenesis.

An Imaging Measurement Study of Normal Knee Parameters in Southeast China.

BACKGROUND: Imaging measurement of distal femur and proximal tibia has been the hot point in the research of total knee arthroplasty and prosthesis development, which is an important treatment for patients with advanced knee joint disease. This study retrospectively investigated the digital imaging measurement of normal knee parameters in southeast China and evaluated their clinical value. METHODS: From February 2010 to May 2014, and in accordance with the inclusion criteria, a total of 677 knees (334 female knees and 343 male knees) were categorized into 3 age groups. Clinical and digital imaging data, including the distal Femoral Condyle Diameter (FCD), Tibial Plateau Diameter (TPD), the distance between the medial tibial plateau and fibular head (DPF), tibiofemoral valgus angle, distal femoral valgus angle, Proximal Tibia (PT) varus angle and the angle from femoral condyle to tibial perpendicular (FT), were measured by using AutoCAD 10.0 software. All measured variables were statistically analyzed by SPSS statistical software (version 18.0). RESULTS: Data are presented as the mean ± standard deviation. The normal female and male femoral condyle diameter was (7.69 ± 0.46) cm and (8.68 ± 0.55) cm, while the normal female and male tibial plateau diameter was (7.66 ± 0.46) cm and (8.60 ± 0.55) cm, respectively. The normal female and male DPF was (0.76 ± 0.36) cm and (0.79 ± 0.36) cm. For females and males, the tibiofemoral valgus angle and distal femoral valgus angle were (3.89 ± 2.20) ° and (3.29 ± 2.12) °, (9.03 ± 2.18) ° and (8.25 ± 2.20) °. As the two methods to measure tibial plateau varus angle, PT angle of normal female and male was (4.29 ± 1.86) ° and (4.84 ± 2.23) °, while the normal female and male FT angle was (5.34 ± 1.95) ° and (5.52 ± 2.07) °. Based on the data obtained, we found significant differences between the two genders in terms of the femoral condyle diameter and tibial plateau diameter in all age groups (P < 0.01). The DPF parameter showed an obvious difference between the young group and the middle-aged group (P < 0.05), and no significant difference was observed between the sides and genders (P > 0.05). The distal femoral valgus angle showed statistical differences between genders in the left side of the young group and middle-aged group (P < 0.05), while angle PT and FT showed no significant difference (P > 0.05). CONCLUSION: A large number of knee measurements was obtained, and a local knee database was developed in this study. Imaging measurement prior to total knee arthroplasty is clinically important for increasing the accuracy and long-term efficacy of total knee arthroplasty. These data can also provide useful information for knee surgery and sports medicine as well as prosthesis development.

Semi-enclosed bays serve as hotspots for black carbon burial: A case study in Jiaozhou Bay, western Yellow Sea.

The provenance of black carbon (BC) and its role in affecting contaminant cycling in both the atmosphere and aquatic environments have been extensively studied. However, the fate and cycling dynamics of BC, particularly in marine environments, are poorly understood. Herein, soot BC was determined in the semi-enclosed Jiaozhou Bay to examine the seasonal variability, residence timescale in seawater, and settling flux to sediments, together with particle-reactive 234Th. Soot BC ranged from 0.39 to 4.26 µmol-C L-1. On average, spring produced the highest value of 1.88 ± 0.31 µmol-C L-1, followed by winter (1.59 ± 0.18 µmol-C L-1), summer (0.94 ± 0.09 µmol-C L-1), and autumn (0.90 ± 0.09 µmol-C L-1). The seasonality of soot BC was similar to the activity concentration of particulate 234Th (i.e., 234ThP). The close relationships between soot BC and 234ThP (p < 0.01) provide the basis for the application of 234Th to trace the fate of soot BC. Based on the 234Th deficit with respect to 238U, the residence times of soot BC were estimated to be 41 ± 6 d and 36 ± 5 d for May-August and August-November, respectively. The shorter residence times of soot BC than that of seawater indicated that soot BC delivered to Jiaozhou Bay settled in the local sediments. Furthermore, soot BC concentrations were higher in the inflow seawater from the Yellow Sea than the outflow water from Jiaozhou Bay, implying a net input of soot BC from the Yellow Sea to Jiaozhou Bay. The soot BC fluxes were 0.266 ± 0.035 mmol-C m-2 d-1 and 0.0472 ± 0.0065 mmol-C m-2 d-1 for May-August and August-November, respectively. From the bay-scale perspective, Jiaozhou Bay had buried 0.101 ± 0.010 Gg of soot BC each year. These results indicate that the semi-enclosed Jiaozhou Bay acts as an effective trap for soot BC and particle-reactive contaminants.

Microalloying Design of Biodegradable Mg-2Zn-0.05Ca Promises Improved Bone-Implant Applications.

Mg and its alloys have been comprehensively studied and show huge potential for clinical orthopedic applications. However, balancing the mechanical strength and corrosion resistance of alloys is still a challenge. In light of this, micro-level contents of Zn and Ca were added to pure Mg to fabricate a Mg-2Zn-0.05Ca microalloy to expectedly enhance the mechanical strength and concurrently improve the corrosion resistance. The characteristics of the rolled Mg-2Zn-0.05Ca microalloy were explored using optical microscopy, X-ray diffraction, and tensile tests. The corrosion behavior and mechanical strength loss were explored using electrochemical and immersion tests. The effects of the microalloy extract on the proliferation, adhesion, and osteogenic differentiation of MC3T3-E1 cells were systematically studied. Moreover, implantations were done in femoral condyles of rabbits to study the degradation properties, osteogenic effect, mechanical strength loss, and biosafety of the microalloy. The ultimate tensile strength and yield strength of the rolled microalloy were found to be significantly elevated to 257 ± 2.74 and 237.6 ± 8.29 MPa, respectively. The microalloy showed a stable and gradual strength loss during degradation, both in vivo and in vitro. Concurrently, the microalloy exhibited improved corrosion resistance ability and especially, in vivo, the rolled microalloy exhibited a comparable degradation rate to that of rolled pure Mg within the initial 12 weeks of implantation. Additionally, the microalloy promoted osteogenesis, both in vitro and in vivo, and no short- and long-term toxicities of the microalloy were observed in rabbits. This study suggested that the rolled Mg-2Zn-0.05Ca microalloy effectively balanced the mechanical strength and corrosion resistance and showed potential application as bone implants.

Association of HSS score and mechanical alignment after primary TKA of patients suffering from constitutional varus knee that caused by combined deformities: a retrospective study.

For pre-operative osteoarthritis (OA) patients with varus knee, previous studies showed inconsistent results. Therefore, we conducted this study to better identify the association of Hospital for Special Surgery (HSS) score and mechanical alignment. 44 patients (51 knees) with constitutional varus knee caused by combined deformities (LDFA (lateral distal femoral angle) > 90°and MPTA (medial proximal tibial angle) < 85°)) were selected and analyzed with a mean follow-up period of 14 months after total knee arthroplasty (TKA). From January 2015 to December 2016, patients were collected consecutively after primary TKA. After filtering, fifty-one knees (44patients) were analyzed with a mean follow-up period of 14 months. All patients were divided into two groups based on post-operative hip-knee-ankle (HKA) acute angle: varus mechanical alignment (VMA) group (HKA < - 3°) and neutral mechanical axis (NMA) group (- 3° ≤ HKA ≤ 3°). 30 knees were included in the NMA group, and 21 knees in the VMA group. Comparisons of HSS between NMA group and VMA group were performed. After adjusting for age and Body Mass Index (BMI) confounders, Compared with NMA group, the HSS score in VMA group decreased by 0.81 units (95% CI, - 3.37 to 1.75) p = 0.5370). For pre-operative constitutional varus knee caused by combined deformities in chinese populations, no significant association between post-operative lower limb mechanical alignment and HSS score was found.

Evaluation of a rabbit model of adjacent intervertebral disc degeneration after fixation and fusion and maintenance in an upright feeding cage.

Purpose: To establish an animal model of adjacent intervertebral disc degeneration by performing spinal fixation and fusion after percutaneous needle puncture and removal of the intervertebral disc or percutaneous needling of the vertebral body without removal of the intervertebral disc. Methods: We established a model of adjacent intervertebral disc degeneration after spinal fixation and fusion of rabbits maintained in upright feeding cages. Twenty-five healthy New Zealand rabbits were used. In the experimental group, the L3-4 intervertebral disc was percutaneously punctured with an 18-G needle under fluoroscopic guidance. Once degeneration occurred, the L3-4 disc was excised, and interbody fusion was performed. The changes in the adjacent intervertebral discs were observed periodically via X-ray and MRI. In the control group, the L3 vertebral body was percutaneously needled with an 18-G needle under fluoroscopic guidance. The changes in the adjacent intervertebral discs were observed on X-ray and MRI at 4, 8, and 12 weeks after puncture in both groups. At 12 weeks postoperatively, the animals were euthanized, and the histopathologic changes of the adjacent intervertebral discs were assessed using hematoxylin-eosin and TdT-mediated dUTP nick end labeling (TUNEL) staining. The mRNA and protein expressions of aggrecanase-1 were measured by real-time quantitative PCR and Western blot analysis. The product of aggrecan degradation, Aggrecan ARGxx, was measured by Western blot analysis. Results: The degeneration of the intervertebral discs in the adjacent segments in the experimental group increased over time. The mRNA and protein expressions of aggrecanase-1 and the expression of Aggrecan ARGxx in the experimental group were significantly increased after puncture, fixation, and fusion (P<0.05). The adjacent intervertebral disc sections had a significantly lower cell density and significantly higher TUNEL-positive cell rate in the experimental group than the control group (P<0.05). Conclusion: The results suggest that the occurrence of intervertebral disc degeneration in adjacent segments may begin with the degeneration of the punctured intervertebral disc.

Netrin-1 regulates ERK1/2 signaling pathway and autophagy activation in wear particle-induced osteoclastogenesis.

BACKGROUND: Artificial joint replacement surgery is often accompanied by osteolysis induced aseptic loosening around the prosthesis. Wear particles from joint replacement are thought to be one of the main factors leading to local inflammation and osteolysis at the prosthesis site. The aim of this study was to investigate the molecular mechanism of osteoclast formation and dissolution induced by wear particles and the potential roles of Netrin-1, the ERK1/2 pathway and autophagy activation in this process. METHODS: The messenger RNA levels in cells and tissues were detected with real-time quantitative PCR. The western blotting was used to detect the expression of proteins. A CCK-8 kit was used to detect the viability of RAW 264.7 cells. Moreover, an air pouch model of bone resorption was established. Immunohistochemistry was used to detect the expression of TRAP and Netrin-1 in rat bone tissue. Cell culture supernatants were collected in the rat air pouch model of bone resorption, and the levels of RANKL and OPG were detected with enzyme-linked immunosorbent assay. The protein levels of TRAP and Netrin-1 in bone tissue were examined by immunohistochemistry. RESULTS: Titanium wear particles induced osteoclast formation and autophagy activation. Moreover, blocking autophagy suppressed the osteoclastogenesis after exposure to wear particles in vitro. The activation of the ERK1/2 pathway and the overexpression of Netrin-1 were both found to play important roles in osteoclastogenesis mediated by autophagy. Moreover, 3-MA effectively decreased the secretion of proinflammatory cytokines mediated by wear particles. CONCLUSION: Blockade of autophagy inhibits the osteoclastogenesis and inflammation induced by wear particles, thus potentially providing novel treatment strategies for abnormal osteoclastogenesis and aseptic prosthesis loosening induced by wear particles.

A persistent increase in primary productivity east off Hainan Island (northwestern South China Sea) over the last decades as inferred from sediment records.

Sediment cores were analyzed from the continental shelf of the northwestern South China Sea aiming to understand the change history of primary productivity and provide insights into key changes of environmental conditions in this region over the past ~100 years. Multiple proxies including stable carbon isotopic composition (δ13C) of sedimentary organic matter, diatom abundance and biogenic silica burial flux were applied along with 210Pb chronology. Notably, these independent evidences consistently point to a steady increase of primary production in this region only after ~1960s. We propose that increasing atmospheric deposition due to dramatically enhanced human activities especially from China supplies essential nitrogen nutrients to the N-poor region and probably acts a major reason for the observed enhancement of marine primary production. Our study provides insights into better understanding how human perturbation may have profoundly impacted biogeochemical cycling in marginal seas in the last decades.

Inhibition of cellular communication network factor 1 (CCN1)-driven senescence slows down cartilage inflammaging and osteoarthritis.

OBJECTIVE: To explore the role of cellular communication network factor 1 (CCN1) in cartilage inflammaging and osteoarthritis (OA) pathogenesis in the isolated primary human chondrocytes in vitro, cartilage explants ex vivo, and a pre-clinical mice model. METHODS: Recombinant human CCN1 stimulation and small interfering RNA inhibition were conducted in human chondrocytes. The RNA was extracted to quantify catabolic targets and pro-inflammatory genes and the proteins were probed with specific antibodies. IL-1ß and IL-6 were monitored by ELISA. IHC was performed to evaluate important hypertrophic hallmarks and catabolic markers. The effects of Tanshinone IIA on chondrocytes were investigated in both time-dependent and dose-dependent processes. Cartilage explants were cultured in growth medium and further treated with Tanshinone IIA. The intra-articular injection was performed in 13 months old C57BL/6J mice. Safranin O and fast green staining were performed to evaluate the histological change of cartilage followed by a semi-quantitative analysis using the OARSI scoring system. RESULTS: RNA and protein levels of CCN1 increased in an age-dependent manner compared to young donors. Increased CCN1 expression was also found in the damaged area compared to the non-lesion area which correlated with the advanced pathological change in human OA. The overexpression of CCN1 promoted chondrocytes senescence, while the down-regulation of CCN1 by small interfering RNA reduced CCN1 production and limited inflammation secretion suggesting that CCN1 was a possible novel target to intervene OA. Inhibition of CCN1 by using Tanshinone IIA could reduce SASP components in a dose- and time-dependent manner. Additionally, our data showed that Tanshinone IIA was able to preserve articular cartilage integrity, suppress CCN1 production, and inhibit SASP factors in human cartilage explants and in aged mice model. CONCLUSION: This study showed that CCN1 signaling aggravated cartilage inflammaing and matrix degradation. Collectively, our findings showed new insight into repurposing Tanshinone IIA for slowing down OA advancement in human and mice by inhibiting the CCN1 axis.