Overloaded axial stress activates the Wnt/ß-Catenin pathway in nucleus pulposus cells of adult degenerative scoliosis combined with intervertebral disc degeneration.

BACKGROUND: Intervertebral disc degeneration (IVDD) is the initiating factor of adult degenerative scoliosis (ADS), and ADS further accelerates IVDD, creating a vicious cycle. Nevertheless, the role of the Wnt/ß-Catenin pathway in ADS combined with IVDD (ADS-IVDD) remains a mystery. Accordingly, this study was proposed to investigate the effect of axial stress on the Wnt/ß-Catenin pathway in nucleus pulposus cells (NPCs) isolated from DS-IVDD patients. METHODS: Normal NPCs (N-NPC) were purchased and the NPCs of young (25-30 years; Y-NPC) and old (65-70 years; O-NPC) from ADS-IVDD patients were primary cultured. After treatment of NPC with overloaded axial pressure, CCK-8 and Annexin V-FITC kits were applied for detecting proliferation and apoptosis of N-NPC, Y-NPC and O-NPC, and western blotting was performed to assess the expression of Wnt 3a, ß-Catenin, NPC markers and apoptosis markers (Bax, Bcl2 and Caspase 3). RESULTS: N-NPC, Y-NPC and O-NPC were mainly oval, polygonal and spindle-shaped with pseudopods, and the cell morphology tended to be flattened with age. N-NPC, Y-NPC and O-NPC were capable of synthesizing proteoglycans and expressing the NPC markers (Collagen II and Aggrecan). Notably, the expression of Wnt 3a, ß-Catenin, Collagen II and Aggrecan was reduced in N-NPC, Y-NPC and O-NPC in that order. After overload axial stress treatment, cell viability of N-NPC and Y-NPC was significantly reduced, and the percentage of apoptosis and expression of Wnt 3a and ß-Catenin were significantly increased. CONCLUSIONS: Overloaded axial pressure activates the Wnt/ß-Catenin pathway to suppress proliferation and facilitate apoptosis of NPC in ADS-IVDD patients.

A breakdown in microglial metabolic reprogramming causes internalization dysfunction of α-synuclein in a mouse model of Parkinson's disease.

BACKGROUND: The α-synuclein released by neurons activates microglia, which then engulfs α-synuclein for degradation via autophagy. Reactive microglia are a major pathological feature of Parkinson's disease (PD), although the exact role of microglia in the pathogenesis of PD remains unclear. Transient receptor potential vanilloid type 1 (TRPV1) channels are nonselective cation channel protein that have been proposed as neuroprotective targets in neurodegenerative diseases. METHODS: Using metabolic profiling, microglia energy metabolism was measured including oxidative phosphorylation and aerobic glycolysis. The mRFP-GFP-tagged LC3 reporter was introduced to characterize the role of TRPV1 in microglial autophagy. α-synuclein preformed fibril (PFF) TRPV1flox/flox; Cx3cr1Cre mouse model of sporadic PD were employed to study the capacity of TRPV1 activation to attenuate neurodegeneration process. RESULTS: We found that acute exposure to PFF caused microglial activation as a result of metabolic reprogramming from oxidative phosphorylation to aerobic glycolysis via the AKT-mTOR-HIF-1α pathway. Activated microglia eventually reached a state of chronic PFF-tolerance, accompanied by broad defects in energy metabolism. We showed that metabolic boosting by treatment with the TRPV1 agonist capsaicin rescued metabolic impairments in PFF-tolerant microglia and also defects in mitophagy caused by disruption of the AKT-mTOR-HIF-1α pathway. Capsaicin attenuated phosphorylation of α-synuclein in primary neurons by boosting phagocytosis in PFF-tolerant microglia in vitro. Finally, we found that behavioral deficits and loss of dopaminergic neurons were accelerated in the PFF TRPV1flox/flox; Cx3cr1Cre mouse model of sporadic PD. We identified defects in energy metabolism, mitophagy and phagocytosis of PFF in microglia from the substantia nigra pars compacta of TRPV1flox/flox; Cx3cr1Cre mice. CONCLUSION: The findings suggest that modulating microglial metabolism might be a new therapeutic strategy for PD.

CHI3L1 promotes Staphylococcus aureus-induced osteomyelitis by activating p38/MAPK and Smad signaling pathways.

AIMS: Staphylococcus aureus (S. aureus) is the most common causative bacterial pathogen involved in promoting infection-induced osteomyelitis, a disease resulting in severe bone degradation. In this study, we aimed to identify the mechanism behind inhibition of osteoclast survival and differentiation by CHI3L1, a lectin previously reported to regulate S. aureus-induced osteomyelitis. MAIN METHODS: The role of CHI3L1 in osteoclast survival, proliferation, and differentiation was studied ex vivo using primary human bone marrow derived stem cells (HBMSCs) and transducing them with lentiviral expression vectors or shRNA knockdown constructs. Cell apoptosis was analyzed by flow cytometry using annexin V-fluorescein isothiocyanate/propidium iodide staining. Cell proliferation was assessed using alkaline phosphatase, Alcian Blue, and TRAP staining. The qRT-PCR was used to measure mRNA levels of osteoclast maturation markers, and western blotting was used to analyze protein expression. An in vivo murine model for osteomyelitis and microcomputed tomography analyses of infected femurs were used to study the effects of CHI3L1 on bone erosion. KEY FINDINGS: Overexpression of CHI3L1 significantly reduced HBMSC cell viability, proliferation, and differentiation, and knockdown improved these effects in the presence of S. aureus infection. More specifically, CHI3L1 constitutively activated the p38/MAPK pathway to promote apoptosis. Furthermore, CHI3L1 induced activation of the Smad pathway by promoting phosphorylation of Smad-1/5 proteins. Finally, overexpression of CHI3L1 significantly induced bone erosion upon S. aureus infection in a murine osteomyelitis model, and knockdown of CHI3L1 significantly alleviated this effect. SIGNIFICANCE: CHI3L1 played a vital role in osteoblast differentiation and proliferation by regulating the p38/MAPK and Smad signaling pathways to promote S. aureus-induced osteomyelitis.

Dynamic Perfusion Culture of Human Outgrowth Endothelial Progenitor Cells on Demineralized Bone Matrix In Vitro.

BACKGROUND The aim of this study was to investigate the proliferation, differentiation, and tube formation of human outgrowth endothelial progenitor cells (OECs) cultured with porous demineralized bone matrix (DBM) under a dynamic perfusion system in vitro. MATERIAL AND METHODS OECs were isolated, expanded, characterized, eGFP-transfected and seeded on DBM scaffold and cultured under static or dynamic perfusion conditions, and continuously observed under fluorescence microscope. DBM scaffolds were harvested on day six for RT-PCR and western blot assay to analyze the mRNA and protein expression level of CD34, VE-cadherin, and VEGF. Scanning electron microscope (SEM) was used to observe the tube formation of OECs seeded on DBM scaffolds. RESULTS The results showed the cell density of OECs on DBM was higher when exposed to shear stress generated by a dynamic perfusion system. Shear stress also markedly increased the expression level of VE-cadherin and VEGF and decreased the expression of CD34, at both mRNA and protein levels. SEM showed that the shear-stressed OECs formed tube-like structures inside the pores of DBM scaffolds. CONCLUSIONS A dynamic perfusion system can be used as an innovative method for the rapid vascularization in tissue engineering, which can accelerate the proliferation and differentiation of OECs and the vascularization of implanted scaffolds.

A narrative review of the progress in the treatment of knee osteoarthritis.

Background and Objective: The pathogenesis of osteoarthritis (OA) involves a variety of complex mechanisms, including genetic, mechanical, metabolic, and inflammatory factors. There is evidence that inflammatory factors, abnormal chondrocyte apoptosis, and extracellular matrix degradation are closely associated with the occurrence and development of OA. The best treatment for OA is still controversial, but intra-articular injection is safer and more effective than non-surgical treatments, such as physical therapy and oral analgesics. This study sought to explore the mechanism, benefits, and adverse reactions of commonly used intra-articular injection therapy in the treatment of knee osteoarthritis (KOA). Methods: We analyzed the safety and adverse reactions of intra-articular injection in patients with KOA, and summarized the results. Key Content and Findings: Six weeks of the corticosteroid injection contributed to improve the symptoms of OA patients in short time. However, their symptoms did not improve significantly after this period. Using corticosteroids for a long time may result in oxidative stress, leading to adverse reactions, such as cartilage toxicity, and accelerate the progress of OA. Due to its high frequency, the local injection of hyaluronic acid can result in more adverse reactions when compared with the corticosteroids. Due to the lack of standardized factors for platelet-rich plasma (PRP) preparation, leukocyte-rich or leukocyte-free variants may be produced. Adverse reactions include injection-site pain, joint stiffness. Conclusions: Thus, it is necessary to promote further clinical trials to promote the clinical application of PRP.

A method for extracting fetal ECG based on EMD-NMF single channel blind source separation algorithm.

Nowadays, detecting fetal ECG using abdominal signal is a commonly used method, but fetal ECG signal will be affected by maternal ECG. Current FECG extraction algorithms are mainly aiming at multiple channels signal. They often assume there is only one fetus and did not consider multiple births. This paper proposed a single channel blind source separation algorithm to process single abdominal acquired signal. This algorithm decomposed single abdominal signal into multiple intrinsic mode function (IMF) utilizing empirical mode decomposition (EMD). Correlation matrix of IMF was calculated and independent ECG signal number was estimated using eigenvalue method. Nonnegative matrix was constructed according to determined number and decomposed IMF. Separation of MECG and FECG was achieved utilizing nonnegative matrix factorization (NMF). Experiments selected four channels man-made signal and two channels ECG to verify correctness and feasibility of proposed algorithm. Results showed that the proposed algorithm could determine number of independent signal in single acquired signal. FECG could be extracted from single channel observed signal and the algorithm can be used to solve separation of MECG and FECG.