CARTILAGE最新文献

ObjectiveThe aim was to investigate starch-gelatin hydrogels as scaffolds for chondrogenesis and compare these with other materials currently in use regarding cell retention and growth.MethodsTwo variants of starch-gelatin-scaffolds and one chitosan-based scaffold were fabricated by casting and freeze-drying. The resulting materials were analyzed with respect to physicochemical and mechanical properties, cut to size, and seeded with human articular chondrocytes. Cell retention and proliferation were evaluated at 1, 14, and 42 days of culturing. Extracellular matrix production was analyzed by histo- and immunohistochemistry. Comparisons were made with that of commercially available hyaluronan- (Hyalofast^®) and collagen-based (ChondroGide^®) scaffolds, and synthesized chitosan hydrogels.ResultsThe starch-gelatin materials exhibited highly porous structures stabilized by hydrogen bonding, with swelling behavior similar to native cartilage and favorable mechanical handling properties. Despite differences in initial cell retention, all materials except chitosan supported robust cell growth, reaching similar levels after 14 days. No significant changes were observed between 14 and 42 days with the exception of Hyalofast^® showing decreased cell number. Chitosan-supported cell growth was more linear over the culture period, but resulted in only half the cell number by day 42 compared with the other materials. Without cells, Hyalofast and one variant of the starch/gelatin hydrogel degraded before day 42. starch/gelatin scaffolds showed collagen I, II, and aggrecan deposition.ConclusionStarch-gelatin scaffolds displayed favorable mechanical properties, supported cell growth comparable to commercial scaffolds, and promoted deposition of cartilage-specific extracellular matrix, highlighting their chondrogenic potential.

ObjectiveAngiogenesis plays a crucial role in osteoarthritis (OA) by promoting inflammatory cell invasion, supporting neo-innervation and joint tissue fibrosis, and contributing to structural damage and pain. Thrombospondin-3 (THBS-3) is highly expressed in OA cartilage. However, the mechanisms responsible for upregulation of THBS-3 in OA are unclear.DesignOA chondrocytes and a collagen-induced osteoarthritis (CIOA) mouse model were used as in vitro and in vivo models, respectively. THBS-3 was used to treat chondrocytes in vitro and in vivo. To explore the mechanism of THBS-3 in chondrocytes treatment, we pretreated chondrocytes with a THBS-3 inhibitor and assessed cartilage metabolic function and then analyzed related indicators of vascularization and chondrofibrosis.ResultsProteomics revealed higher THBS-3 expression in the cartilage of CIOA mice than in that of normal mice. Compared with those from healthy individuals, chondrocytes from OA patients presented significantly increased protein expression of THBS-3. In both in vivo and in vitro experiments, THBS-3 promoted matrix metalloproteinase-13 and disintegrin and metalloprotease with thrombospondin-5, suppressed aggrecan, and promoted the vascularization and chondrofibrosis in dysfunctional chondrocytes from osteoarthritic chondrocytes. THBS-3 activated the transforming growth factor-beta (TGF-β) signaling pathway. The pretreatment of OA chondrocytes with a TGF-β inhibitor before THBS-3 exposure reversed these changes.ConclusionTHBS-3 promotes the angiogenesis and fibrosis of chondrocytes by activating the TGF-β/Smad2/3 signaling pathway.

BackgroundDiclofenac etalhyaluronate (DF-HA, SI-613/ONO-5704) is a conjugate of hyaluronic acid (HA) and diclofenac (DF), and its intra-articular injection is widely used for the treatment of osteoarthritis in Japan. While novel mechanisms of cartilage protection by DF-HA have been identified, a comprehensive analysis of the biological responses unique to DF-HA has not yet been conducted.DesignWe used an RNA sequencing (RNA-seq) method to comprehensively analyze gene expression in the knee joint cartilage of arthritic rats and cytokine-stimulated chondrocytes. For the mechanistic analysis of DF-HA, genes that were downregulated or upregulated by DF-HA, HA, or DF were extracted. Pathway analysis was then performed on genes that specifically varied with DF-HA treatment.ResultsIn the cartilage of rats with collagen-induced arthritis, treatment with DF-HA, but not DF or HA, suppressed the extracellular matrix (ECM) remodeling pathway and promoted the parathyroid hormone/parathyroid hormone-related peptide receptor-mediated pathway, which regulates chondrocyte differentiation and bone/cartilage development. In cytokine-stimulated chondrocytes, DF-HA similarly suppressed the ECM remodeling pathway; specifically, gene expression changes in IGFBP4, MMP10, MMP13, and TIMP1 were consistent with those observed in vivo.ConclusionRNA-seq analysis of cartilage in arthritic rats and cytokine-stimulated chondrocytes provided molecular mechanistic insights, indicating that DF-HA treatment induced cartilage protection through the suppression of ECM remodeling.

ObjectiveArtificial intelligence offers opportunities for timesaving assessments of multiple pathologies in large magnetic resonance imaging (MRI) data sets in knee osteoarthritis (KOA). This study evaluated their prevalence within pre-defined clinical phenotypes and their predictive value for knee replacement (KR).DesignBaseline MRIs (n = 8,667) from the Osteoarthritis Initiative were analyzed using a machine-learning (ML) algorithm. The presence of pathologies (menisci, anterior cruciate, medial collateral ligaments, cartilage, etc.) was assessed in previously identified phenotypic clusters (a post-traumatic, metabolic, and age-defined phenotype). The value of both, cluster allocation and joint pathology for KR prediction was evaluated using supervised ML models and time-dependent receiver operating characteristic curves.ResultsCompared to the population average, the metabolic cluster had a higher prevalence of cartilage lesions, while the post-traumatic one had more medial meniscal damage. Random forest models showed the best prediction (area under the curve 0.837, test set at 2 years). The top predictors for KR were meniscal position (relative to the border of the tibial plateau), severe joint effusion, medial femorotibial cartilage lesions, and metabolic phenotype. These features defined patients at high risk of KR with an estimated KR rate at 5 years of 10% vs 3% in the high- and low-risk groups based on a predictive risk score including all analyzed structures.ConclusionsThis ML-enabled assessment of multiple MRI pathologies in a large KOA data set highlights the importance of meniscal pathologies and markers of inflammation, in addition to cartilage assessments and clinical information for patient stratification and improved prediction of KOA progression to KR.

ObjectivesDamaged articular cartilage cannot regenerate spontaneously. Chondrocyte therapy, the current treatment of choice, requires laboratory expansion, necessitating two surgical procedures. Adding a second cell type to intraoperatively isolated chondrocytes enables single-stage chondrocyte-based repair strategies and has shown promise in vitro. The benefit of this strategy in vivo and in the clinic has not yet been comprehensively assessed. This systematic review assesses the efficacy of cartilage repair by co-implantations of chondrocytes with other cell types over all available in vivo studies.DesignMedline, Embase and Cochrane databases were searched for studies on co-implantations of chondrocytes with other cell types for hyaline cartilage repair. For each study, extracted data were tabulated and reporting quality and risk of bias were assessed. Studies were categorized based on their control groups for qualitative synthesis.ResultsThe search yielded 48 studies across 46 publications: 44 animal studies (26 ectopic and 18 orthotopic) and four clinical trials. Ectopic studies scored poorly on reporting quality and bias, while orthotopic studies were only moderately better. Twenty-seven of 27 experiments with chondrocyte-only controls demonstrated synergistic cartilage formation through co-implantation. Cartilage formation by co-implantations was also found in 15 of 17 experiments without such control and in four clinical trials.ConclusionCartilage repair by articular chondrocytes is improved by adding a second cell type, like mesenchymal stromal cells. This is likely caused by the second cell types' stimulatory support. Other second cell types and chondrocyte sources have shown promising results too. This is encouraging for further clinical exploration of single-stage co-implantation strategies.

IntroductionThis study explores the interplay between miRNA-140-5p expression and mechanical stress in cartilage within joint biomechanics.MethodsTibial plateau specimens, CT data, and mechanical parameters were obtained from healthy and OA donors. Twenty-four mice, including 12 miRNA-140-5p knockdown (MUT-group) and 12 wild-type (WT group), underwent anterior-cruciate-ligament-transection (ACLT) or sham-operation (SHAM). Finite element analysis, 3D simulation, CT scans, immunohistochemical staining, and fluorescence in situ hybridization were conducted. Primary chondrocytes with or without miRNA-140-5p agomir were loaded mechanically and analyzed by RT-qPCR, Western blot, and phalloidin staining.ResultsThe mechanical coupling unit comprised articular cartilage and subchondral bone, with cartilage apparent compressive modulus linked to the trabecular bone structure (P < 0.05). Healthy-joint samples and low-stress regions in animal samples exhibited high miRNA-140-5p expression (P < 0.05) and low RhoA expression. OA or high-stress regions showed the opposite trend (P < 0.05). MiRNA-140-5p knockdown increased joint loading in mice. In vitro, miRNA-140-5p overexpression reduced RhoA and cytoskeletal remodeling, maintaining chondrocyte mechano-responsiveness.ConclusionsOur study reveals a link between mechanical stress and miRNA-140-5p, implying its role in maintaining joint mechanical homeostasis. These findings enhance understanding of biomechanical-molecular interplay, though further studies are needed to assess therapeutic potential in osteoarthritis.

ObjectiveOsteoarthritis (OA) is characterized by articular cartilage erosion, pathological subchondral bone changes, and signs of synovial inflammation and pain. We previously identified p[63-82], a bone morphogenetic protein 7 (BMP7)-derived bioactive peptide that attenuates structural cartilage degeneration in the rat medial meniscal tear-model for posttraumatic OA. This study aimed to evaluate the cartilage erosion-attenuating activity of p[63-82] in a different preclinical model for OA (anterior cruciate ligament transection-partial medial meniscectomy [anterior cruciate ligament transection (ACLT)-pMMx]). The disease-modifying action of the p[63-82] was followed-up in this model for 5 and 10 weeks.DesignSkeletally mature male Lewis rats underwent ACLT-pMMx surgery. Rats received weekly intra-articular injections with either saline or 500 ng p[63-82]. Five and 10 weeks postsurgery, rats were sacrificed, and subchondral bone characteristics were determined using microcomputed tomography (µCT). Histopathological evaluation of cartilage degradation and Osteoarthritis Research Society International (OARSI)-scoring was performed following Safranin-O/Fast Green staining. Pain-related behavior was measured by incapacitance testing and footprint analysis.ResultsHistopathological evaluation at 5 and 10 weeks postsurgery showed reduced cartilage degeneration and a significantly reduced OARSI score, whereas no significant changes in subchondral bone characteristics were found in the p[63-82]-treated rats compared to the saline-treated rats. ACLT-pMMx-induced imbalance of static weightbearing capacity in the p[63-82] group was significantly improved compared to the saline-treated rats at weeks 5 postsurgery. Footprint analysis scores in the p[63-82]-treated rats demonstrated improvement at week 10 postsurgery.ConclusionsWeekly intra-articular injections of p[63-82] in the rat ACLT-pMMx posttraumatic OA model resulted in reduced degenerative cartilage changes and induced functional improvement in static weightbearing capacity during follow-up.

ObjectiveOsteoarthritis (OA) is the most common arthritic disease in humans. Nevertheless, the pathogenic mechanism of OA remains unclear. This study aimed to explore that heat-shock transcription factor 1 (HSF1) facilitated interleukin-1 beta (IL-1β) chondrocyte injury by increasing Notch1 O-linked N-acetylglucosamine (O-GlcNAc) modification level.DesignHuman chondrocytes were incubated with 5 ng/ml interleukin-1 beta (IL-1β) for 24 h to establish OA cell model. The messenger RNA (mRNA) or protein expressions were assessed using reverse transcription-quantitative polymerase chain reaction, western blot, or immunofluorescence. Chondrocyte viability was examined by Cell Counting Kit-8 assay. Enzyme-linked immunosorbent assay was employed to detect the secretion levels of interleukin-6 (IL-6) and interleukin-8 (IL-8). Immunoprecipitation was adopted to detect Notch1 O-GlcNAc modification level. The interaction between HSF1 and epidermal growth factor-like (EGF) domain-specific O-GlcNAc transferase (EOGT) promoter was analyzed by dual-luciferase reporter gene and chromatin immunoprecipitation assays.ResultsHerein, our results demonstrated that HSF1, EOGT, Notch1, and Notch1 intracellular domain (NICD1) expressions in chondrocytes were markedly increased by IL-1β stimulation. EOGT elevated Notch1 expression in IL-1β-treated chondrocytes by increasing Notch1 O-GlcNAc modification level. EOGT silencing reduced IL-1β-induced chondrocyte inflammatory injury. In addition, HSF1 knockdown relieved IL-1β-induced chondrocyte inflammatory injury. Molecular interaction experiment proved that HSF1 transcriptionally activated EOGT expression in IL-1β-treated chondrocytes.ConclusionsHSF1 promoted IL-1β-induced inflammatory injury in chondrocytes by increasing EOGT-mediated glycosylation of Notch1.

ObjectiveThis proof-of-concept study investigated an improved cell-based injection therapy combining mesenchymal stem cells (MSCs) and meniscus cells (MCs) to support superior meniscus allograft repopulation and early revival compared to injecting MSCs alone.DesignIn this controlled laboratory study, frozen meniscus allograft samples were injected vertically with a cell suspension containing different ratios of MSCs and MCs or control (lactated ringers) and cultured for 28 days. Samples were analyzed weekly for cell viability, migration, and metabolism using histological and biochemical assays. Tissue medium was analyzed for matrix metalloproteinase (MMP) expression using zymography.ResultsCellular repopulation of frozen allografts injected with different cell suspensions was validated by immunohistochemistry. Significant higher DNA content was evidenced in grafts treated with suspensions of MCs or MC:MSC (1:4 ratio). Cell metabolic activity was significantly different between all treated groups and control group after 1 week. Allografts injected with MCs showed significantly more cell proliferation than injections with MSCs. MMP2 activity was detected in medium of all grafts cellularized with MCs with or without MSCs. Scanning electron microscopy (SEM) analysis showed resolution of the needle puncture, but not in the control group. Cell labeling of MCs upon injection of mixed MC:MSC suspensions revealed a gradual increase in the cell ratio.ConclusionsThe findings of this study establish that injection of MCs with or without MSCs enhances the cellularity of meniscus allograft to support early graft revival and remodeling.

Purpose/AimThis study aimed to assess the effectiveness of joint lavage in managing knee osteoarthritis (OA) by evaluating its effect on pain relief, inflammatory markers, cartilage-degrading enzymes, and oxidative stress.MethodsSeventy patients with Kellgren-Lawrence grade 2 or 3 knee OA were selected for this single-center study. Joint lavage was performed, and pain and function were measured using the visual analog scale (VAS) and Western Ontario and McMaster Universities Osteoarthritis Index (WOMAC) scores at baseline and 24 weeks postintervention. Synovial fluid samples were collected at baseline, before lavage, and 24 weeks postintervention. Samples were stored at -80°C and analyzed in batches to minimize variability. At the time of analysis, the samples were thawed and evaluated for levels of proinflammatory cytokines, interleukin-1β (IL-1β), interleukin-6 (IL-6), and tumor necrosis factor alpha (TNF-α), matrix metalloproteinase-3 (MMP-3), and total oxidant status (TOS), and oxidative stress index (OSI).ResultsPostintervention, VAS, and WOMAC scores significantly decreased (P < 0.001), with 100% achieving the minimal clinically important difference (MCID). Patient acceptable symptom state (PASS) rates varied: VAS (80%), WOMAC pain (50%), function (81.4%), and total (84.3%). Cytokine levels (IL-1β, IL-6, TNF-α) and MMP-3 significantly decreased (P < 0.001), along with TOS and OSI. Baseline TNF-α, IL-6, and IL-1β levels were significantly correlated with improvements in VAS and WOMAC scores. Moderate correlations were observed between reductions in IL-6/TNF-α and improvements in VAS/WOMAC. No significant associations were found between confounders and outcomes.ConclusionsJoint lavage resulted in marked pain relief and functional improvement while significantly reducing inflammatory markers, cartilage-degrading enzymes, and oxidative stress.