Tension regulates the cartilage phenotypic expression of endplate chondrocytes through the α-catenin/actin skeleton/Hippo pathway.

The study aimed to investigate the regulatory mechanism of intracellular tension signaling in endplate chondrocytes and its impact on extracellular matrix synthesis. Human endplate chondrocytes were subjected to tension load using Flexcell FX-5000™, and changes in phenotype, morphology, and the expression of Hippo signaling pathway and α-Catenin were assessed through various techniques. Through the overexpression of YAP and inhibition of α-Catenin, the study clarified the intracellular tension signaling pathway and its regulation of extracellular matrix synthesis in endplate cartilage. In vitro-cultured human endplate chondrocytes significantly suppressed phenotype-related genes and proteins, accompanied by distinct changes in cytoskeleton morphology. Tension activation resulted in the substantial activation of the Hippo pathway, increased phosphorylation of YAP, and reduced nuclear translocation of YAP. YAP overexpression alleviated the inhibitory effect of tension on extracellular matrix synthesis in endplate chondrocytes. Tension also upregulated the expression of α-Catenin in endplate chondrocytes, which was attenuated by inhibiting α-Catenin expression, thereby reducing the impact of tension on cytoskeletal morphology and YAP nuclear translocation. Taken together, the α-Catenin/actin skeleton/Hippo-coupled network is a crucial signaling pathway for tension signaling in endplate chondrocytes, providing potential therapeutic targets for the treatment of endplate cartilage degeneration.

Clinical significance of redundant nerve roots in patients with lumbar spinal stenosis undergoing oblique lumbar interbody fusion combined with percutaneous internal fixation.

BACKGROUND: While there have been previous studies on the surgical efficacy of patients with redundant nerve roots (RNRs), a persistent issue is that some patients continue to experience redundancy even after surgery. Furthermore, the clinical significance of RNRs remains unclear. Notably, there is a lack of research regarding RNRs within the context of oblique lumbar interbody fusion (OLIF) combined with percutaneous internal fixation. Therefore, the primary objective of this study is to investigate the correlation between RNRs and clinical outcomes following OLIF combined with percutaneous internal fixation. METHODS: Eighty-seven patients diagnosed with lumbar spinal stenosis (LSS) who underwent single-segment OLIF combined with percutaneous internal fixation were categorized into three groups. Group 1 comprised patients with positive RNRs both before and after the operation. Group 2 included patients with positive RNRs preoperatively but negative RNRs postoperatively. Group 3 consisted of patients with consistently negative RNRs before and after the operation. Comprehensive patient data were collected, including operation time, intraoperative blood loss, and any recorded complications. Radiographic parameters, both pre- and post-operative, were assessed, encompassing the number of stenosis segments, disc height (DH), lumbar lordotic angle, dural sac cross-sectional area, and the placement of the fusion cage. Furthermore, the Visual Analogue Scale was applied to gauge back and leg pain, while the Oswestry Disability Index was employed to appraise daily living activities. A comparative analysis was carried out among the three patient groups. RESULTS: In this study, all 87 LSS patients successfully underwent surgery. Among them, 35 patients (40.2%) showed preoperative MRI assessment indicating positive RNRs. In the postoperative MRI assessment, 14 of these patients maintained positive RNRs status, and they were grouped into Group 1. The remaining 21 patients saw a transition to negative RNRs status and were included in Group 2. Among the 52 patients who had preoperative MRI assessments showing negative RNRs, their postoperative RNRs status remained negative, forming Group 3. All patients received follow-up, which ranged from 8 to 18 months, and no complications occurred during this period. In this study, the postoperative efficacy and parameters such as DH and Dural Sac CSA significantly improved compared to preoperative values for all 87 patients. Patients with preoperative RNRs had more stenosis segments, smaller dural sac CSA, and more severe symptoms. In all three groups, postoperative efficacy scores significantly improved compared to preoperative scores. Group 2 patients had their fusion cages placed more in the middle, while Group 1 patients had their fusion cages more anteriorly located. Group 2 patients exhibited greater recovery in dural sac CSA postoperatively compared to Group 1 patients. Additionally, Group 2 patients had better ODI efficacy scores compared to Group 1 patients. CONCLUSIONS: Irrespective of the presence or absence of RNRs, patients experienced improvement after undergoing OLIF combined with percutaneous internal fixation. Preoperative RNRs appear to be linked to multi-segmental lumbar spinal stenosis, a reduction in dural sac CSA, and symptom severity. Patients with negative postoperative RNRs demonstrated better treatment efficacy. Furthermore, the placement of the fusion cage appears to have a significant impact on postoperative efficacy and RNRs outcomes.

Study on the safety and effectiveness of low-dose vs. regular-dose fondaparinux in preventing venous thromboembolism prophylaxis following total knee arthroplasty.

Background: This study aims to evaluate the effectiveness and safety of low-dose (1.5 mg) fondaparinux for venous thromboembolism (VTE) prophylaxis in patients post-total knee arthroplasty (TKA). Methods: We retrospectively identified 314 patients who carried out the primary TKAs and received fondaparinux for VTE chemoprophylaxis between July 2020 and December 2021. A total of 141 TKA patients were excluded according to the exclusion criteria. Two groups of patients were established: the low-dose group included 84 patients who injected 1.5 mg of fondaparinux, and the regular-dose group included 89 patients who injected 2.5 mg of fondaparinux. The pre-operative blood analysis and coagulation assays were performed. The surgical time, the incidence of symptomatic VET, blood loss, wound complication, bleeding, drainage, and mortality of patients were determined and assessed. Results: The pre-operative blood analysis, body mass index, sex, age, and coagulation assays of patients in both groups were comparable. In terms of symptomatic pulmonary embolism and deep vein thrombosis, there was no significant difference (variation) between the two groups. However, patients in both groups showed a substantial difference in terms of blood loss, drain volume, wound complication, and transfusion rate. Conclusion: In prevention of VET in patients post-TKA, low-dose fondaparin is as effective as conventional dose fondaparinux. A significant decrease in blood loss, post-surgical transfusion rates, and wound complications were detected in patients given low-dose fondaparinux compared to those receiving regular-dose fondaparinux.

Corrigendum: Coating of manganese functional polyetheretherketone implants for osseous interface integration.

[This corrects the article DOI: 10.3389/fbioe.2023.1182187.].

Coating of manganese functional polyetheretherketone implants for osseous interface integration.

Polyetheretherketone (PEEK) has been used extensively in biomedical engineering and it is highly desirable for PEEK implant to possess the ability to promote cell growth and significant osteogenic properties and consequently stimulate bone regeneration. In this study, a manganese modified PEEK implant (PEEK-PDA-Mn) was fabricated via polydopamine chemical treatment. The results showed that manganese was successfully immobilized on PEEK surface, and the surface roughness and hydrophilicity significantly improved after surface modification. Cell experiments in vitro demonstrated that the PEEK-PDA-Mn possesses superior cytocompatibility in cell adhesion and spread. Moreover, the osteogenic properties of PEEK-PDA-Mn were proved by the increased expression of osteogenic genes, alkaline phosphatase (ALP), and mineralization in vitro. Further rat femoral condyle defect model was utilized to assess bone formation ability of different PEEK implants in vivo. The results revealed that the PEEK-PDA-Mn group promoted bone tissue regeneration in defect area. Taken together, the simple immersing method can modify the surface of PEEK, giving outstanding biocompatibility and enhanced bone tissue regeneration ability to the modified PEEK, which could be applied as an orthopedic implant in clinical.

A dual-functional PEEK implant coating for anti-bacterial and accelerated osseointegration.

Polyetheretherketone (PEEK) has been widely applied in biomedical engineering. However, the unsatisfactory bioactivity essentially limits the clinical application of PEEK. In this study, a simply immersing method was proposed to fabricate a dual-functional PEEK with antibacterial properties and enhanced bone integration. Firstly, the surface of PEEK was modified with a polydopamine (PDA) coating by incubating at dopamine solution. Afterward, the PEEK-PDA was modified with manganese (Mn) and silver (Ag) ions by the soaking method to fabricate the PEEK-PDA-Mn/Ag. The physicochemical capabilities of PEEK-PDA-Mn/Ag were further explored in the ions release, wettability, morphology, and element distributions. PEEK-PDA-Mn/Ag obviously accelerated the adhesion and distribution of MC3T3-E1 cells, indicating favorable biosafety in vitro. Meanwhile, the osteogenic properties of PEEK-PDA-Mn and PEEK-PDA-Mn/Ag were proved by the increased expression of osteogenic genes, alkaline phosphatase (ALP), and mineralization in vitro. Additionally, the wide antibacterial capabilities of PEEK-PDA-Mn/Ag were proved in both Staphylococcus aureus (S. aureus) and Escherichia coli (E. coli) in vitro. Furthermore, the PEEK-PDA-Mn/Ag was antibacterial with capability in enhancing osseointegration in vivo. Overall, the simply immersing method can modify the surface of PEEK, giving the bioactivity, biocompatibility, and antibacterial ability to the composited PEEK, which could be applied as an orthopedic implant in clinical.

DNA methyltransferase 3 beta mediates the methylation of the microRNA-34a promoter and enhances chondrocyte viability in osteoarthritis.

Osteoarthritis (OA) is characterized by destruction of articular cartilage with an imbalance between synthesis and degradation of extracellular matrix (ECM). In the current study, we explored the role of microRNA-34a (miR-34a) and the behind epigenetic mechanism in the degradation of ECM in OA. Using miRNA-based microarray analysis, we found that miR-34a was overexpressed in cartilage tissues of OA patients relative to patients with acute traumatic amputations. Moreover, its expression was positively correlated with the ECM degradation and inflammation. Mechanistically, miR-34a targeted MCL1, and possible target genes of miR-34a were enriched in the PI3K/AKT pathway. Furthermore, DNMT3B inhibited miR-34a by promoting miR-34a methylation. Functional experiments using CCK-8, flow cytometry, Safranin O staining, RT-qPCR, ELISA, Western blot, and HE staining revealed that miR-34a inhibitor suppressed ECM degradation and inflammatory response of chondrocytes and cartilage tissues. By contrast, downregulation of DNMT3B and MCL1 reversed the repressive effects of miR-34a inhibitor in vitro and in vivo. Altogether, our findings establish that silencing of miR-34a by DNMT3B could effectively reduce chondrocyte ECM degradation and inflammatory response in mice by targeting MCL1 and mediating the downstream PI3K/AKT pathway. This present study revealed that miR-34a knockdown might develop a novel intervention for OA treatment.

[Expression and significance of Wnt/ß-catenin signaling pathway in vitro natural degenerationmodel of endplate chondrocytes in rats].

OBJECTIVE: To explore the expression and significance of Wnt/ß-catenin signaling pathway in the natural degeneration of endplate chondrocytes in rats. METHODS: Endplate chondrocytes extracted from lumbar vertebrae were divided into control (P2 cell), naturally passaged (P5 cell) and wnt signaling pathway inhibition groups. The morphology of endplate chondrocytes was observed by inverted microscope. Hematoxylin and eosin (HE) and toluidine blue stains were used to identify their phenotypes. Type II collagen marker, SOX-9 and aggrecan genes were detected by reverse transcription-polymerase chain reaction (RT-PCR) to verify the degeneration model. Based on this model, the changes of ß-catenin were detected by RT-PCR and Western blot. Laser confocal microscopy was used to detect the expression and localization of ß-catenin within endplate chondrocytes. RESULTS: With natural passaging, endplate cartilage cells appeared spindle-shaped and gradually lost chondrocytic phenotypes. The levels of type II collagen (P5/P2 = 0.11, P = 0.003 9), SOX-9 (P5/P2 = 0.58, P = 0.016 8) and aggrecan (P5/P2 = 0.32, P = 0.004 6) significantly reduced; ß-catenin (P5/P2 = 1.62, P = 0.008 2) significantly increased. ß-catenin was down-regulated (XAV-939/P5 = 0.23, P = 0.001 7) in inhibition group. And type II collagen (XAV-939/P5 = 2.60, P = 0.018 0), SOX-9 (XAV-939/P5 = 1.47, P = 0.038 2) and aggrecan (XAV-939/P5 = 2.56, P = 0.004 1) significantly increased. ß-catenin had a higher expression and obviously entered into nuclear transcription in P5 generation and decreased in inhibition group. CONCLUSION: ß-catenin plays an important role in the in vitro degeneration of endplate chondrocytes. There are great potentials for protecting endplate cartilage degeneration by regulating the Wnt/ß-catenin signaling pathway.

The evidence and the possible significance of autophagy in degeneration model of human cervical end-plate cartilage.

The aim of this study was to observe autophagy in chondrocytes from degenerative human cervical vertebral end-plates and to investigate the significance of variations in autophagy in the degeneration of cervical vertebral end-plate chondrocytes. Cartilage end-plates were obtained from 48 inpatients admitted to hospital between February 2011 and August 2012. The patients were divided into the control group (n=17) with cervical vertebral fracture or dislocation and the cervical spondylosis group (n=31) with cervical spondylotic myelopathy. End-plate chondrocytes were isolated via enzyme digestion and then cultured in vitro. The cells were stained with toluidine blue and hematoxylin-eosin (H&E). A laser scanning confocal microscope and monodansylcadaverine (MDC) were used to reveal autophagy in the end-plate chondrocytes. Reverse transcription polymerase chain reaction (RT-PCR) was used to detect mRNA expression of type II collagen and aggrecan. Western blotting was conducted to detect LC3 proteins. The chondrocytes isolated from the degenerative human cervical end-plates were cultured successfully in vitro. The morphology of the cells from the cervical spondylosis group tended to exhibit changes in spindle morphology compared with the control group. Autophagic bodies were stained with MDC. LC3 proteins were visible in the intracellular and perinuclear regions under the laser scanning confocal microscope. The mRNA expression levels (relative to those of ß-actin) of aggrecan (0.715±0.194) and type II collagen (0.628±0.254) in the cervical spondylosis group were markedly decreased compared with those in the control group (0.913±0.254 and 0.845±0.186, respectively; both P<0.05). The LC3-II/LC3-I ratio was observed to be significantly reduced in the cervical spondylosis group by Western blot analysis. Autophagy has an important role in human cervical disc degeneration. The regulation of autophagy may prevent disc degeneration in cartilage end-plate cells.

[Autophagy expression and its significance in chondrocytes from a degenerate model of human cervical endplate].

OBJECTIVE: To explore the autophagy expression and examine its significance in chondrocytes in a degenerate model of human cervical vertebrae endplate. METHODS: Cartilage endplates were obtained from 48 hospitalized patients with cervical vertebral fracture or dislocation at our hospital between February 2012 to August 2012. They were divided into cervical spondylosis group with cervical spondylotic myelopathy (n = 31) and control group (n = 17).Endplate chondrocytes were isolated by enzyme digestion and cultured in vitro. The cells were stained with toluidine blue and hematoxylin and eosin; laser scanning confocal microscope and monodansylcadaverine (MDC) were used to observe autophagy in endplate chondrocytes; reverse transcription-polymerase chain reaction (RT-PCR) was used to detect the mRNA expression of typeIIcollagen and aggrecan and Western blot for the protein of LC3. RESULTS: A degenerative cell model of human cervical endplate chondrocytes was established successfully in vitro. Compared with the common group, the cellular morphologies of degenerative group showed spindle changes. Autophagic body was stained with MDC.Intracellular and perinuclear LC3 protein was detected by laser confocal microscopy. Compared with the control group, the mRNA expressions of aggrecan (0.715 ± 0.194) and typeII collagen (0.628 ± 0.254) markedly decreased (0.845 ± 0.186,0.913 ± 0.254, P < 0.05) and LC3-II/LC3-I declined in cervical spondylosis group. CONCLUSION: Autophagy plays an important pathogenic role in the process of human cervical disc degeneration. And regulating its expression may improve disc degeneration in endplate cartilage cells.

[Change of autophagy in endplate chondrocytes of rats during aging process].

OBJECTIVE: To explore the expression and significance of autophagy in endplate cartilage of rats during aging process. METHODS: The end-plate chondrocytes were isolated from 3, 6 and 12-month SD rats respectively. And the natural culture and rapamycin groups were assigned. Alizarin red staining was used to observe the morphological changes of cells. And RT-PCR was employed to detect the expressions of type II collagen, proteoglycan, SOX-9 and matrix metalloproteinase (MMP-13). The expressions of Beclin-I and LC3 were detected by reverse transcription-polymerase chain reaction (RT-PCR) and Western blot. The rate of autophagy was observed by monodansylcadaverine (MDC) staining and methyl thiazolyl tetrazolium (MTT) for cell survival rate. RESULTS: Alizarin red staining showed that cells might reflect the process of intervertebral disc degeneration. The expressions of polysaccharide, Sox-9, type II collagen, Beclin-1 and LC3 in endplate chondrocytes significantly decreased with advancing age (P < 0.05). The incidence of autophagy significantly decreased (P < 0.05). The cell viability of each group significantly decreased (P < 0.05). Compared with control group, the cell viability of rapamycin group significantly increased (P < 0.05). CONCLUSION: During aging process, the expressions of autophagy related-gene LC3 and Beclin-1 significantly decrease with the reduced activity of end-plate chondrocyte. And autophagy activity may be correlated with the development and degeneration of intervertebral disc.