RESUMEN
Low back pain (LBP) is largely attributed to intervertebral disc degeneration (IVDD), of which the endplate changes are an important component. However, the alterations in cell fate and properties within the endplates during degeneration remain unknown. Here, we firstly performed the single-cell RNA-sequencing analysis (scRNA-seq) of the cells focusing on degenerative human endplates. By unsupervised clustering of the 8,534 single-cell based on the gene expression, we identified nine distinct cell types. We employed Gene ontology (GO) analysis, Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways analysis, and the single-cell regulatory network inference and clustering (SCENIC) to determine the enriched pathways and transcriptional activities across seven chondrocyte subpopulations. Furthermore, two cell fates of chondrocyte differentiation were found by trajectory analysis, one was enriched in inflammation-related genes, and the other was related to extracellular matrix (ECM). Additionally, the intercellular interactions of macrophages (MA) and chondrocytes, T cells/natural killer cells (T/NK) and chondrocytes were examined by ligand-receptor pairs analysis, showing the important regulative function of FN1 from MA and CD74 from T/NK during endplate degeneration. Overall, our findings provide novel perspectives on the endplate degeneration at the single-cell level and a whole-transcriptome size.
Asunto(s)
Diferenciación Celular , Condrocitos , Degeneración del Disco Intervertebral , Análisis de Secuencia de ARN , Análisis de la Célula Individual , Humanos , Degeneración del Disco Intervertebral/genética , Degeneración del Disco Intervertebral/patología , Análisis de la Célula Individual/métodos , Condrocitos/metabolismo , Condrocitos/patología , Diferenciación Celular/genética , Perfilación de la Expresión Génica , Femenino , Masculino , Redes Reguladoras de Genes , Persona de Mediana Edad , Macrófagos/metabolismo , Adulto , Disco Intervertebral/patología , Disco Intervertebral/metabolismoRESUMEN
Intervertebral disc (IVD) degeneration damaging the extracellular matrix (ECM) of IVDs is the main cause of spine-associated disorders. Degenerative disc disease (DDD) is a multifaceted disorder, where environmental factors, inflammatory cytokines and catabolic enzymes act together. DDD starts typically due to imbalance between ECM biosynthesis and degradation within IVDs, especially through unbalanced degradation of aggrecan and collagen II in nucleus pulposus (NP). Current treatment approaches are primarily based on conservative or surgical therapies, which are insufficient for biological regeneration. The disintegrins and metalloproteinases with thrombospondin motifs (ADAMTSs) and matrix metalloproteinases (MMPs) are the key proteolytic enzymes for degradation of aggrecan and collagens. Previously, high expression levels of ADAMTS4, ADAMTS5, MMP3 and MMP13, which are accompanied with low levels of aggrecan and collagen II, were demonstrated in degenerative human NP cells. Moreover, self-complementary adeno-associated virus type 6 (scAAV6) mediated inhibitions of ADAMTS4 and ADAMTS5 by RNA-interference (RNAi) could specifically enhance aggrecan level. Thus, MMPs are apparently the main degrading enzymes of collagen II in NP. Furthermore, scAAV6-mediated inhibitions of MMP3 and MMP13 have not yet been investigated. Therefore, we attempted to enhance the level of collagen II in degenerative NP cells by scAAV6-RNAi-mediated inhibitions of MMP3 and MMP13. MRI was used to determine preoperative grading of IVD degeneration in patients. After isolation and culturing of NP cells, cells were transduced with scAAV6-shRNAs targeting MMP3 or MMP13; and analysed by fluorescence microscopy, FACS, MTT assay, RT-qPCR, ELISA and western blotting. scAAV6-shRNRs have no impact on cell viability and proliferation, despite high transduction efficiencies (98.6%) and transduction units (1383 TU/Cell). Combined knockdown of MMP3 (92.8%) and MMP13 (90.9%) resulted in highest enhancement of collagen II (143.2%), whereby treatment effects were significant over 56 days (p < 0.001). Conclusively, scAAV6-RNAi-mediated inhibitions of MMP3 and MMP13 help to progress less immunogenic and enduring biological treatments in DDD.
Asunto(s)
Proteína ADAMTS4 , Degeneración del Disco Intervertebral , Metaloproteinasa 13 de la Matriz , Metaloproteinasa 3 de la Matriz , Núcleo Pulposo , Metaloproteinasa 3 de la Matriz/metabolismo , Metaloproteinasa 3 de la Matriz/genética , Humanos , Metaloproteinasa 13 de la Matriz/metabolismo , Degeneración del Disco Intervertebral/metabolismo , Degeneración del Disco Intervertebral/terapia , Degeneración del Disco Intervertebral/patología , Degeneración del Disco Intervertebral/genética , Núcleo Pulposo/metabolismo , Núcleo Pulposo/patología , Proteína ADAMTS4/metabolismo , Proteína ADAMTS4/genética , Colágeno Tipo II/metabolismo , Dependovirus/genética , Dependovirus/metabolismo , Proteína ADAMTS5/metabolismo , Proteína ADAMTS5/genética , Interferencia de ARN , Células Cultivadas , Agrecanos/metabolismoRESUMEN
Intervertebral disc degeneration (IDD) is a major cause of discogenic pain, and is attributed to the dysfunction of nucleus pulposus, annulus fibrosus, and cartilaginous endplate (CEP). Osteopontin (OPN), a glycoprotein, is highly expressed in the CEP. However, little is known on how OPN regulates CEP homeostasis and degeneration, contributing to the pathogenesis of IDD. Here, we investigate the roles of OPN in CEP degeneration in a mouse IDD model induced by lumbar spine instability and its impact on the degeneration of endplate chondrocytes (EPCs) under pathological conditions. OPN is mainly expressed in the CEP and decreases with degeneration in mice and human patients with severe IDD. Conditional Spp1 knockout in EPCs of adult mice enhances age-related CEP degeneration and accelerates CEP remodeling during IDD. Mechanistically, OPN deficiency increases CCL2 and CCL5 production in EPCs to recruit macrophages and enhances the activation of NLRP3 inflammasome and NF-κB signaling by facilitating assembly of IRAK1-TRAF6 complex, deteriorating CEP degeneration in a spatiotemporal pattern. More importantly, pharmacological inhibition of the NF-κB/NLRP3 axis attenuates CEP degeneration in OPN-deficient IDD mice. Overall, this study highlights the importance of OPN in maintaining CEP and disc homeostasis, and proposes a promising therapeutic strategy for IDD by targeting the NF-κB/NLRP3 axis.
Asunto(s)
Inflamasomas , Degeneración del Disco Intervertebral , Macrófagos , Ratones Noqueados , FN-kappa B , Proteína con Dominio Pirina 3 de la Familia NLR , Osteopontina , Transducción de Señal , Adulto , Anciano , Anciano de 80 o más Años , Animales , Femenino , Humanos , Masculino , Ratones , Persona de Mediana Edad , Adulto Joven , Cartílago/patología , Cartílago/metabolismo , Condrocitos/metabolismo , Condrocitos/patología , Inflamasomas/metabolismo , Degeneración del Disco Intervertebral/patología , Degeneración del Disco Intervertebral/metabolismo , Degeneración del Disco Intervertebral/genética , Macrófagos/metabolismo , Ratones Endogámicos C57BL , FN-kappa B/metabolismo , Proteína con Dominio Pirina 3 de la Familia NLR/metabolismo , Proteína con Dominio Pirina 3 de la Familia NLR/genética , Proteína con Dominio Pirina 3 de la Familia NLR/deficiencia , Osteopontina/metabolismo , Osteopontina/deficiencia , Osteopontina/genéticaRESUMEN
Low back pain significantly impacts individuals' quality of life, with intervertebral disc degeneration (IDD) being a primary contributor to this condition. Currently, IDD treatment primarily focuses on symptom management and does not achieve a definitive cure. The cartilage endplate (CEP), a crucial nutrient-supplying tissue of the intervertebral disc, plays a pivotal role in disc degeneration. This review examines the mechanisms underlying CEP degeneration, summarizing recent advancements in understanding the structure and function of CEP, the involvement of various signaling pathways, and the roles of cartilage endplate stem cells (CESCs) and exosomes (Exos) in this process. The aim of this review is to provide a comprehensive reference for future research on CEP. Despite progress in understanding the role of CEP in IDD, the mechanisms underlying CEP degeneration remain incompletely elucidated. Future research poses significant challenges, necessitating further investigations to elucidate the complexities of CEP.
Asunto(s)
Cartílago , Degeneración del Disco Intervertebral , Degeneración del Disco Intervertebral/patología , Degeneración del Disco Intervertebral/metabolismo , Humanos , Cartílago/metabolismo , Cartílago/patología , Animales , Disco Intervertebral/patología , Disco Intervertebral/metabolismo , Exosomas/metabolismo , Células Madre/metabolismo , Células Madre/citología , Células Madre/patología , Transducción de SeñalRESUMEN
INTRODUCTION: The vertebral cartilage endplate (CEP), crucial for intervertebral disc health, is prone to degeneration linked to chronic low back pain, disc degeneration, and Modic changes (MC). While it is known that disc cells express toll-like receptors (TLRs) that recognize pathogen- and damage-associated molecular patterns (PAMPs and DAMPs), it is unclear if CEP cells (CEPCs) share this trait. The CEP has a higher cell density than the disc, making CEPCs an important contributor. This study aimed to identify TLRs on CEPCs and their role in pro-inflammatory and catabolic gene expression. METHODS: Gene expression of TLR1-10 was measured in human CEPs and expanded CEPCs using quantitative polymerase chain reaction. Additionally, surface TLR expression was measured in CEPs grouped into non-MC and MC. CEPCs were stimulated with tumor necrosis factor alpha, interleukin 1 beta, small-molecule TLR agonists, or the 30 kDa N-terminal fibronectin fragment. TLR2 signaling was inhibited with TL2-C29, and TLR2 protein expression was measured with flow cytometry. RESULTS: Ex vivo analysis found all 10 TLRs expressed, while cultured CEPCs lost TLR8 and TLR9 expression. TLR2 expression was significantly increased in MC1 CEPCs, and its expression increased significantly after pro-inflammatory stimulation. Stimulation of the TLR2/6 heterodimer upregulated TLR2 protein expression. The TLR2/1 and TLR2/6 ligands upregulated pro-inflammatory genes and matrix metalloproteases (MMP1, MMP3, and MMP13), and TLR2 inhibition inhibited their upregulation. Endplate resorptive capacity of TLR2 activation was confirmed in a CEP explant model. CONCLUSIONS: The expression of TLR1-10 in CEPCs suggests that the CEP is susceptible to PAMP and DAMP stimulation. Enhanced TLR2 expression in MC1, and generally in CEPCs under inflammatory conditions, has pro-inflammatory and pro-catabolic effects, suggesting a potential role in disc degeneration and MC.
Asunto(s)
Receptor Toll-Like 2 , Receptores Toll-Like , Humanos , Receptor Toll-Like 2/metabolismo , Receptor Toll-Like 2/genética , Receptores Toll-Like/metabolismo , Receptores Toll-Like/genética , Cartílago/metabolismo , Cartílago/patología , Masculino , Femenino , Persona de Mediana Edad , Disco Intervertebral/metabolismo , Disco Intervertebral/patología , Inflamación/patología , Inflamación/genética , Inflamación/metabolismo , Regulación de la Expresión Génica , Adulto , Degeneración del Disco Intervertebral/genética , Degeneración del Disco Intervertebral/metabolismo , Degeneración del Disco Intervertebral/patología , Anciano , Transducción de SeñalRESUMEN
OBJECTIVE: To systematically review relevant animal models of disk degeneration induced through the endplate injury pathway and to provide suitable animal models for exploring the intrinsic mechanisms and treatment of disk degeneration. DESIGN: PubMed, Web of Science, Cochrane and other databases were searched for literature related to animal models of disk degeneration induced by the endplate injury pathway from establishment to August 2024, and key contents in the literature were screened and extracted to analyze and evaluate each type of animal model using the literature induction method. RESULTS: Fifteen animal experimental studies were finally included in the literature, which can be categorized into direct injury models and indirect injury models, of which direct injury models include transvertebral injury models and transpedicular approach injury models, and indirect injury models include endplate ischemia models and vertebral fracture-induced endplate injury models. The direct injury models have a minimum observation period of 2 months and a maximum of 32 wk. All direct injury models were successful in causing disk degeneration, and the greater the number of interventions, the greater the degree of disk degeneration caused. The observation period for the indirect injury models varied from 4 wk to 70 wk. Of the 9 studies, only one study was unsuccessful in inducing disk degeneration, and this was the first animal study in this research to attempt to intervene on the endplate to cause disk degeneration. CONCLUSION: The damage to the direct injury model is more immediate and controllable in extent and can effectively lead to disk degeneration. The indirect injury models do not directly damage the endplate structure, making it easier to observe the physiological and pathological condition of the endplate and associated structures of the disk. None of them can completely simulate the corresponding process of endplate injury-induced disk degeneration in humans, and there is no uniform clinical judgment standard for this type of model. The most appropriate animal model still needs further exploration and discovery.
Asunto(s)
Modelos Animales de Enfermedad , Degeneración del Disco Intervertebral , Disco Intervertebral , Degeneración del Disco Intervertebral/etiología , Degeneración del Disco Intervertebral/patología , Animales , Disco Intervertebral/lesiones , Disco Intervertebral/patología , HumanosAsunto(s)
Degeneración del Disco Intervertebral , Disco Intervertebral , Degeneración del Disco Intervertebral/metabolismo , Degeneración del Disco Intervertebral/patología , Humanos , Disco Intervertebral/metabolismo , Disco Intervertebral/patología , Disco Intervertebral/efectos de los fármacos , Animales , Lípidos , Metabolismo de los Lípidos/efectos de los fármacosRESUMEN
BACKGROUND: Intervertebral disc (IVD) degeneration is a multifactorial pathological process resulting in the dysregulation of IVD cell activity. The catabolic shift observed in IVD cells during degeneration leads to increased inflammation, extracellular matrix (ECM) degradation, aberrant intracellular signaling and cell loss. Importantly, these pathological processes are known to be interconnected and to collectively contribute to the progression of the disease. MicroRNAs (miRNAs) are known as strong post-transcriptional regulators, targeting multiple genes simultaneously and regulating numerous intracellular pathways. Specifically, miR-155-5p has been of particular interest since it is known as a pro-inflammatory mediator and contributing factor to diseases like cancer and osteoarthritis. This study investigated the role of miR-155-5p in IVD degeneration with a specific focus on inflammation and mechanosensing. METHODS: Gain- and loss-of-function studies were performed through transfection of human Nucleus pulposus (NP) and Annulus fibrosus (AF) cells isolated from degenerated IVDs with miR-155-5p mimics, inhibitors or their corresponding non-targeting control. Transfected cells were then subjected to an inflammatory environment or mechanical loading. Conditioned media and cell lysates were collected for phosphorylation and cytokine secretion arrays as well as gene expression analysis. RESULTS: Increased expression of miR-155-5p in AF cells resulted in significant upregulation of interleukin (IL)-8 cytokine secretion during cyclic stretching and a similar trend in IL-6 secretion during inflammation. Furthermore, miR-155-5p mimics increased the expression of the brain-derived neurotrophic factor (BDNF) in AF cells undergoing cyclic stretching. In NP cells, miR-155-5p gain-of-function resulted in the activation of the mitogen-activated protein kinase (MAPK) signaling pathway through increased phosphorylation of p38 and p53. Lastly, miR-155-5p inhibition caused a significant increase in the anti-inflammatory cytokine IL-10 in AF cells and the tissue inhibitor of metalloproteinases (TIMP)-4 in NP cells respectively. CONCLUSION: Overall, these results show that miR-155-5p contributes to IVD degeneration by enhancing inflammation through pro-inflammatory cytokines and MAPK signaling, as well as by promoting the catabolic shift of AF cells during mechanical loading. The inhibition of miR-155-5p may constitute a potential therapeutic approach for IVD degeneration and low back pain.
Asunto(s)
Inflamación , Degeneración del Disco Intervertebral , MicroARNs , MicroARNs/genética , MicroARNs/metabolismo , Degeneración del Disco Intervertebral/genética , Degeneración del Disco Intervertebral/metabolismo , Degeneración del Disco Intervertebral/patología , Humanos , Inflamación/genética , Inflamación/patología , Inflamación/metabolismo , Núcleo Pulposo/metabolismo , Núcleo Pulposo/patología , Masculino , Soporte de Peso , Persona de Mediana Edad , Femenino , Anillo Fibroso/metabolismo , Anillo Fibroso/patologíaRESUMEN
BACKGROUND: Reduction of inflammatory damage and inhibition of nucleus pulposus (NP) apoptosis are considered to be the main effective therapy idea to reverse the intervertebral disc degeneration (IDD) and alleviate the chronic low back pain. The adenosine A2A receptor (A2AR), as a member of G protein-coupled receptor families, plays an important role in the anti-inflammation and relieving pain. So far, the impact of A2AR on IDD therapy is unclear. The aim of this study was to explore the role of Adenosine A2A receptor (A2AR) in the intervertebral disc degeneration (IDD) and clarify potential mechanism. MATERIALS AND METHODS: IL-1ß and acupuncture was used to establish IDD model rats. A2AR agonist CGS-21680 and A2AR antagonist SCH442416 were used to investigate the therapeutical effects for IDD. Histological examination, western blotting analysis and RT-PCR were employed to evaluate the the association between A2AR and cyclic adenosine monophosphate (cAMP)/protein kinase A (PKA) pathway. RESULTS: A2AR activity of the intervertebral disc tissues was up-regulated in feedback way, and cAMP, PKA and CREB expression were also increased. But in general, IL-1ß-induced IDD promoted the significant up-regulation the expression of inflammatory factors. The nucleus pulposus (NP) inflammation was exacerbated in result of MMP3 and Col-II decline through activating NF-κB signaling pathway. A2AR agonist CGS-21680 exhibited a disc protective effect through significantly increasing A2AR activity, then further activated cAMP/PKA signaling pathway with attenuating the release of TNF-α and IL-6 via down-regulating NF-κB. In contrast, SCH442416 inhibited A2AR activation, consistent with lower expression levels of cAMP and PKA, further leading to the acceleration of IDD. CONCLUSIONS: The activation of A2AR can prevent inflammatory responses and mitigates degradation of IDD thus suggest a potential novel therapeutic strategy of IDD.
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Proteínas Quinasas Dependientes de AMP Cíclico , Inflamación , Degeneración del Disco Intervertebral , FN-kappa B , Receptor de Adenosina A2A , Transducción de Señal , Degeneración del Disco Intervertebral/metabolismo , Degeneración del Disco Intervertebral/patología , Degeneración del Disco Intervertebral/tratamiento farmacológico , Animales , Proteínas Quinasas Dependientes de AMP Cíclico/metabolismo , Receptor de Adenosina A2A/metabolismo , FN-kappa B/metabolismo , Transducción de Señal/efectos de los fármacos , Ratas , Inflamación/metabolismo , Masculino , Ratas Sprague-Dawley , Fenetilaminas/farmacología , Núcleo Pulposo/metabolismo , Núcleo Pulposo/patología , Núcleo Pulposo/efectos de los fármacos , AMP Cíclico/metabolismo , Agonistas del Receptor de Adenosina A2/farmacología , Modelos Animales de Enfermedad , Adenosina/análogos & derivadosRESUMEN
AIM: The degradation of the cartilage endplate (CEP) plays a critical role in the initiation and progression of intervertebral disc degeneration (IVDD), a disease closely associated with inflammation and oxidative stress. Naringin (NGN), a flavonoid compound derived from citrus fruits, has been shown to exhibit significant anti-inflammatory and antioxidant properties. This suggests a promising avenue for NGN's application in IVDD therapy. This study aims to elucidate the therapeutic effects and underlying mechanisms of NGN on CEP degeneration, contributing to the formulation of evidence-based treatment strategies for IVDD. METHODS: In vivo, we developed an intervertebral disc degeneration (IVDD) model in mice by excising the bilateral facet joints and surrounding ligaments, and evaluated the effects of naringin using HE staining and Micro-CT analysis. In vitro, endplate chondrocytes were isolated and subjected to TBHP to replicate the IVDD pathological condition. The protective effects of NGN on these cells were confirmed through immunofluorescence, Western Blot, and flow cytometry. RESULTS: In vivo, NGN effectively mitigated IVDD progression and CEP calcification in mice. In vitro, NGN enhanced mitophagy and suppressed NLRP3 inflammasome activation through the SIRT3/FOXO3a/Parkin pathway. Furthermore, NGN safeguarded chondrocytes against apoptosis and calcification triggered by oxidative stress, in addition to mitigating the degradation of the extracellular matrix. However, silencing SIRT3 negated NGN's protective influence on chondrocytes. CONCLUSION: Our study demonstrated that NGN effectively shields chondrocytes from apoptosis and NLRP3 inflammasome activation by facilitating SIRT3-mediated mitophagy. These insights could pave the way for innovative approaches in the prevention and management of IVDD.
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Apoptosis , Condrocitos , Flavanonas , Proteína Forkhead Box O3 , Inflamasomas , Degeneración del Disco Intervertebral , Ratones Endogámicos C57BL , Mitofagia , Proteína con Dominio Pirina 3 de la Familia NLR , Sirtuina 3 , Animales , Proteína con Dominio Pirina 3 de la Familia NLR/metabolismo , Condrocitos/efectos de los fármacos , Condrocitos/metabolismo , Condrocitos/patología , Mitofagia/efectos de los fármacos , Apoptosis/efectos de los fármacos , Inflamasomas/metabolismo , Flavanonas/farmacología , Flavanonas/uso terapéutico , Degeneración del Disco Intervertebral/tratamiento farmacológico , Degeneración del Disco Intervertebral/patología , Sirtuina 3/metabolismo , Ratones , Proteína Forkhead Box O3/metabolismo , Masculino , Modelos Animales de Enfermedad , Ubiquitina-Proteína Ligasas/metabolismo , Células Cultivadas , Transducción de Señal/efectos de los fármacos , Antiinflamatorios/farmacología , Antiinflamatorios/uso terapéuticoRESUMEN
BACKGROUND: Lipid metabolism disorders are associated with degeneration of multiple tissues and organs, but the mechanism of crosstalk between lipid metabolism disorder and intervertebral disc degeneration (IDD) has not been fully elucidated. In this study we aim to investigate the regulatory mechanism of abnormal signal of lipid metabolism disorder on intervertebral disc endplate chondrocyte (EPC) senescence and calcification. METHODS: Human intervertebral disc cartilage endplate tissue, cell model and rat hyperlipemia model were performed in this study. Histology and immunohistochemistry were used to human EPC tissue detection. TMT-labelled quantitative proteomics was used to detect differential proteins, and MRI, micro-CT, safranin green staining and immunofluorescence were performed to observe the morphology and degeneration of rat tail intervertebral discs. Flow cytometry, senescence-associated ß-galactosidase staining, alizarin red staining, alkaline phosphatase staining, DCFH-DA fluorescent probe, and western blot were performed to detect the expression of EPC cell senescence, senescence-associated secretory phenotype, calcification-related proteins and the activation of cell senescence-related signaling pathways. RESULTS: Our study found that the highly expressed oxidized low-density lipoprotein (ox-LDL) and Lectin-like oxidized low-density lipoprotein receptor 1 (LOX-1) in human degenerative EPC was associated with hyperlipidemia (HLP). TMT-labelled quantitative proteomics revealed enriched pathways such as cell cycle regulation, endochondral bone morphogenesis and inflammation. The rat model revealed that HLP could induce ox-LDL, LOX-1, senescence and calcification markers high expression in EPC. Moreover, we demonstrated that ox-LDL-induced EPCs senescence and calcification were dependent on the LOX-1 receptor, and the ROS/P38-MAPK/NF-κB signaling pathway was implicated in the regulation of senescence induced by ox-LDL/LOX-1 in cell model. CONCLUSIONS: So our study revealed that ox-LDL/LOX-1-induced EPCs senescence and calcification through ROS/P38-MAPK/NF-κB signaling pathway, providing information on understanding the link between lipid metabolism disorders and IDD.
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Senescencia Celular , Condrocitos , Degeneración del Disco Intervertebral , Metabolismo de los Lípidos , Lipoproteínas LDL , Receptores Depuradores de Clase E , Degeneración del Disco Intervertebral/metabolismo , Degeneración del Disco Intervertebral/patología , Lipoproteínas LDL/metabolismo , Animales , Humanos , Receptores Depuradores de Clase E/metabolismo , Condrocitos/metabolismo , Condrocitos/patología , Ratas , Masculino , Calcinosis/metabolismo , Calcinosis/patología , Disco Intervertebral/metabolismo , Disco Intervertebral/patología , Modelos Animales de Enfermedad , Femenino , Persona de Mediana Edad , Transducción de Señal , Adulto , Proteómica/métodos , Ratas Sprague-DawleyRESUMEN
This study aims to identify potential targets and regulatory mechanisms of Astragaloside â £ (AS-â £) in treating intervertebral disc degeneration (IDD) through network pharmacology analysis with experimental validation. Lumbar spine instability (LSI) mouse models were first established and treated with AS-â £. Micro-CT, safranin O-fast green staining, IDD score, RT-PCR and immunohistochemistry staining were employed to demonstrate the effect of AS-â £. Network pharmacology was used to predict the signaling pathways and potential targets of AS-â £ in treating IDD. RT-PCR and immunohistochemistry staining were used to elucidate and validate the mechanism of AS-â £ in vivo. Animal experiments showed that AS-â £ maintained disc height and volume, improved matrix metabolism in LSI mice, and restored Col2α1, ADAMTS-5, Aggrecan, and MMP-13 expression in degenerated discs. Network pharmacology analysis identified 32 cross-targets between AS-â £ and IDD, and PPI network analysis filtered out 11 core genes, including ALB, MAPK1, MAPK14 (p38 MAPK), EGFR, TGFBR1, MAPK8, MMP3, ANXA5, ESR1, CASP3, and IGF1. Enrichment analysis revealed that 7 of the 11 core target genes enriched in the MAPK signaling pathway, and AS-â £ exhibited stable binding to them according to molecular docking results. Experimental validation indicated that AS-â £ reversed mRNA levels of 7 core targets in degenerated disc tissues in LSI mice. Immunohistochemistry staining further revealed that AS-â £ treatment mainly depressed IDD-elevated protein levels of EGFR, p38 MAPK and CASP3 in the annulus fibrosus. This study elucidates that AS-â £ alleviates lumbar spine instability-induced IDD in mice, suggesting the mechanism may involve inhibition of the EGFR/MAPK signaling pathway.
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Degeneración del Disco Intervertebral , Farmacología en Red , Saponinas , Triterpenos , Animales , Degeneración del Disco Intervertebral/tratamiento farmacológico , Degeneración del Disco Intervertebral/metabolismo , Degeneración del Disco Intervertebral/patología , Saponinas/farmacología , Saponinas/uso terapéutico , Triterpenos/farmacología , Triterpenos/uso terapéutico , Ratones , Masculino , Modelos Animales de Enfermedad , Transducción de Señal/efectos de los fármacos , Ratones Endogámicos C57BL , Mapas de Interacción de Proteínas , Metaloproteinasa 13 de la Matriz/metabolismo , Metaloproteinasa 13 de la Matriz/genética , Vértebras Lumbares/efectos de los fármacos , Vértebras Lumbares/patología , Vértebras Lumbares/metabolismo , Regulación de la Expresión Génica/efectos de los fármacos , Disco Intervertebral/efectos de los fármacos , Disco Intervertebral/metabolismo , Disco Intervertebral/patologíaRESUMEN
PURPOSE: This study investigated potential use of computed tomography (CT)-based parameters in the lumbar spine as a surrogate for magnetic resonance imaging (MRI)-based findings. METHODS: In this retrospective study, all individuals, who had a lumbar spine CT scan and MRI between 2006 and 2012 were reviewed (n = 198). Disc height (DH) and endplate degeneration (ED) were evaluated between Th12/L1-L5/S1. Statistics consisted of Spearman correlation and univariate/multivariable regression (adjusting for age and gender). RESULTS: The mean CT-DH increased kranio-caudally (8.04 millimeters (mm) at T12/L1, 9.17 mm at L1/2, 10.59 mm at L2/3, 11.34 mm at L3/4, 11.42 mm at L4/5 and 10.47 mm at L5/S1). MRI-ED was observed in 58 (29%) individuals. CT-DH and MRI-DH had strong to very strong correlations (rho 0.781-0.904, p < .001). MRI-DH showed higher absolute values than CT-DH (mean of 1.76 mm). There was a significant association between CT-DH and MRI-ED at L2/3 (p = .006), L3/4 (p = .002), L4/5 (p < .001) and L5/S1 (p < .001). A calculated cut-off point was set at 11 mm. CONCLUSIONS: In the lumbar spine, there is a correlation between disc height on CT and MRI. This can be useful in trauma and emergency cases, where CT is readily available in the lack of an MRI. In addition, in the middle and lower part of the lumbar spine, loss of disc height on CT scans is associated with more pronounced endplate degeneration on MRIs. If the disc height on CT scans is lower than 11 mm, endplate degeneration on MRIs is likely more pronounced. LEVEL AND DESIGN: Level III, a retrospective study.
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Degeneración del Disco Intervertebral , Disco Intervertebral , Vértebras Lumbares , Imagen por Resonancia Magnética , Tomografía Computarizada por Rayos X , Humanos , Vértebras Lumbares/diagnóstico por imagen , Vértebras Lumbares/patología , Imagen por Resonancia Magnética/métodos , Masculino , Femenino , Tomografía Computarizada por Rayos X/métodos , Estudios Retrospectivos , Persona de Mediana Edad , Adulto , Degeneración del Disco Intervertebral/diagnóstico por imagen , Degeneración del Disco Intervertebral/patología , Anciano , Disco Intervertebral/diagnóstico por imagen , Disco Intervertebral/patología , Anciano de 80 o más Años , Adulto JovenRESUMEN
As a common disease, cervical spondylosis (CS) results from the degeneration of the cervical intervertebral disc. However, there are still no effective clinical strategies for the treatment of this disease. Needle-scalpel (Ns), a therapy guided by traditional Chinese medicine theory, alleviates intervertebral disc degradation and is widely used in the clinic to treat CS. Stromal cell-derived factor-1 (SDF-1) and its receptor CXC receptor 4 (CXCR4) in nucleus pulposus cells play an important role in CS onset and development. This study aimed to explore whether Ns can relieve pain and regulate the SDF-1/CXCR4 axis in nucleus pulposus cells to inhibit apoptosis, thereby delaying cervical intervertebral disc degradation in a rat model of CS. It was found that the Ns-treated groups exhibited higher mechanical allodynia scores than the model group, and H&E staining, MRI, and scanning electron microscopy revealed that Ns therapy inhibited intervertebral disc degeneration. Additionally, Ns therapy significantly inhibited increases in the RNA and protein expression levels of SDF-1 and CXCR4. Furthermore, these treatments alleviated the apoptosis of nucleus pulposus cells, which manifested as a decline in the proportion of apoptotic nucleus pulposus cells and inhibition of the decrease in the levels of Bcl-2/Bax. These findings indicated that Ns mitigated CS-induced pain, inhibited the apoptosis of nucleus pulposus cells, and alleviated intervertebral disc degeneration in CS rats. These effects may be mediated by specifically regulating the SDF-1/CXCR4 signaling axis. Based on these findings, we conclude that Ns might serve as a promising therapy for the treatment of CS.
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Apoptosis , Quimiocina CXCL12 , Modelos Animales de Enfermedad , Degeneración del Disco Intervertebral , Núcleo Pulposo , Ratas Sprague-Dawley , Receptores CXCR4 , Animales , Receptores CXCR4/metabolismo , Receptores CXCR4/genética , Quimiocina CXCL12/metabolismo , Apoptosis/efectos de los fármacos , Núcleo Pulposo/metabolismo , Núcleo Pulposo/efectos de los fármacos , Núcleo Pulposo/patología , Degeneración del Disco Intervertebral/metabolismo , Degeneración del Disco Intervertebral/patología , Degeneración del Disco Intervertebral/tratamiento farmacológico , Ratas , Masculino , Vértebras Cervicales , Transducción de Señal/efectos de los fármacos , Espondilosis/metabolismo , Espondilosis/patologíaRESUMEN
Objective: To investigate the relationship between degeneration of cervical intervertebral disc and degeneration of paravertebral muscles[multifidus (MF), cervical semispinalis (SCer), semispinalis capitis (SCap) and splenius capitis (SPL)]. Methods: 82 patients with chronic neck pain were randomly selected, including 43 males and 39 females, with 50.73 0.7.51 years old. All patients were scanned by 3.0T MRI Philips Ingenia performed conventional MRI sequence scanning and fat measurement sequence mDIXON-Quant scanning of cervical. Fat infiltration (FI) and cross-sectional area (CSA) of cervical paravertebral muscle (MF, SCer, SCap and SPL) at central level of C5-6 disc were measured by Philips 3.0T MRI image post-processing workstation. According to Pfirrmann grading system, there was no grade I in the included cases. The number of grade IIr IV cases were n=16, 40, 19 and 7 respectively. CSA and FI of cervical paravertebral muscles were compared with t test or one-way ANOVA, Spearman correlation analysis was used to evaluate the correlation between age, disc degeneration, and CSA, FI of cervical paravertebral muscles, and multiple linear regression analysis was used to analyze the independent influencing factors of CSA and FI. Results: CSA of cervical paravertebral muscles in male patients was significantly higher than that in female patients (all P<0.001), but there was no significant difference in FI (all P>0.05). Age was weakly correlated with CSA of MF+SCer, moderately correlated with CSA of SCap and SPL (r=-0.256, -0.355 and -0.361, P<0.05), weakly correlated with FI of SCap and SPL (r= 0.182 and 0.264, P<0.001), moderately correlated with FI of MF+SCer (r=0.408, P<0.001). There were significant differences in FI with disc degeneration (P<0.001, P=0.028 and P=0.005). Further correlation analysis showed that disc degeneration was strongly correlated with FI of MF+SCer (r=0.629, P<0.001), and moderately correlated with FI of SCap and SPL (r=0.363, P=0.001; r=0.345, P=0.002). Multiple linear regression analysis showed that sex and age were the influencing factors of CSA of SCap and SPL, sex was the independent influencing factor of CSA of MF+SCer, and disc degeneration was the independent influencing factor of FI. Conclusions: Age is negatively correlated with CSA and positively correlated with FI. Disc degeneration was correlated with FI of paravertebral muscles, especially with FI of MF and SCer. Sex and age were the influencing factors of CSA, while disc degeneration was the independent influencing factor of FI.
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Vértebras Cervicales , Degeneración del Disco Intervertebral , Imagen por Resonancia Magnética , Humanos , Masculino , Femenino , Degeneración del Disco Intervertebral/diagnóstico por imagen , Degeneración del Disco Intervertebral/patología , Persona de Mediana Edad , Vértebras Cervicales/diagnóstico por imagen , Vértebras Cervicales/patología , Imagen por Resonancia Magnética/métodos , Adulto , Músculos Paraespinales/diagnóstico por imagen , Músculos Paraespinales/patología , Dolor de Cuello/diagnóstico por imagen , Dolor de Cuello/patología , AncianoRESUMEN
OBJECTIVE: To determine the occurrence of degenerative changes affecting the vertebral column in cats, assess their clinical significance, and determine the occurrence in cats with intervertebral disk herniation compared to other spinal diseases. ANIMALS: 114 client-owned cats. METHODS: Hospital records were retrospectively reviewed for cats with suspected myelopathy that had undergone spinal MRI. Signalment; history; neurological examination; neurolocalization; primary diagnosis; presence, type, and location of intervertebral disk herniation; and presence and location of other degenerative spinal changes (intervertebral disk degeneration [IVDD], spondylosis deformans [SD], end plate changes, dorsal compressions [DC], and foraminal stenosis [FS]) were recorded. RESULTS: 70% of cats showed at least 1 spinal degenerative change. The most common change was IVDD, followed by SD and intervertebral disk protrusion (IVDP), while intervertebral disk extrusion (IVDE), end plate changes, DC, and FS were uncommon to rare. Primary complaint was attributed to a degenerative condition in 22% of cats, including 100% with IVDE, 9% with IVDP, and 43% with degenerative lumbosacral stenosis (DLSS). The occurrence of degenerative spinal changes and number of intervertebral disks affected by IVDD significantly increased with age and body weight. Age was positively correlated with the occurrence of SD and DLSS. Intervertebral disk degeneration, IVDP, SD, DC, and FS were more prevalent in the lumbosacral junction. Cats with IVDD were significantly more likely to show IVDE and IVDP. CLINICAL RELEVANCE: This study revealed that in a population of cats presenting for signs of myelopathy, IVDE was always responsible for the clinical presentation, DLSS was commonly considered incidental, and IVDP was infrequently related to neurological signs.
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Enfermedades de los Gatos , Degeneración del Disco Intervertebral , Desplazamiento del Disco Intervertebral , Imagen por Resonancia Magnética , Estenosis Espinal , Animales , Gatos , Femenino , Masculino , Enfermedades de los Gatos/diagnóstico por imagen , Enfermedades de los Gatos/patología , Relevancia Clínica , Degeneración del Disco Intervertebral/veterinaria , Degeneración del Disco Intervertebral/diagnóstico por imagen , Degeneración del Disco Intervertebral/patología , Desplazamiento del Disco Intervertebral/diagnóstico por imagen , Desplazamiento del Disco Intervertebral/patología , Desplazamiento del Disco Intervertebral/veterinaria , Imagen por Resonancia Magnética/veterinaria , Estudios Retrospectivos , Estenosis Espinal/diagnóstico por imagen , Estenosis Espinal/patología , Estenosis Espinal/veterinariaRESUMEN
OBJECTIVE: Multiple-level Intervertebral disc degeneration (IDD) in patients with lumbar disc herniation (LDH) is related to postoperative re-herniation and low back pain. Although many investigators believed that there is an interdependence between paraspinal muscles degeneration and IDD, few studies focused on the fatty infiltration of paraspinal muscles on single- and multiple-level IDD in patients with LDH. This study aims to investigate the difference on the fatty infiltration of paraspinal muscles between single- and multiple-levels IDD in patients with LDH. and to explore in patients with LDH whether fatty infiltration is a potential risk factor for multiple-level IDD. METHODS: This study was conducted as a retrospective observational analysis of 82 patients with LDH from January 1, 2020 to December 30, 2020 in our hospital were enrolled. Twenty-seven cases had single-level IDD (Group A), and 55 cases had multiple-level IDD (Group B). We measured the mean computed tomography (CT) density value of the paraspinal muscles, including multifidus (MF), erector spinae (ES) and psoas muscle (PM) at each disc from L1 to S1. Subgroups were set to further analyze the odds ratio (OR) of fatty infiltration of paraspinal muscles in different sex and BMI groups. We measured sagittal angles and analyzed the relationships between these angles and IDD. Finally, we use logistic regression, adjusted for other confounding factors, to investigate whether fatty infiltration is an independent risk factor for multi-level IDD. RESULTS: The average age in multi-level IDD (51.40 ± 15.47 years) was significantly higher than single-level IDD (33.37 ± 7.10 years). The mean CT density value of MF, ES and PM in single-level IDD was significantly higher than multi-level IDD (all ps < 0.001). There was no significant difference of the mean value of angles between the two groups. No matter being fat (body mass index [BMI] > 24.0 kg/m2) or normal, patients with low mean muscle CT density value of MF and ES are significantly easier to suffer from multiple-level IDD. In the pure model, the average CT density value of the MF, ES and PM is all significantly associated with the occurrence of multi-IDD. However, after adjusting for various confounding factors, only the OR of the average CT density value for MF and ES remains statistically significant (OR = 0.810, 0.834, respectively). CONCLUSIONS: In patients with LDH, patients with multiple-level IDD have more severe fatty infiltration of MF and ES than those with single-level IDD. Fatty infiltration of MF and ES are independent risk factors for multiple-level IDD in LDH patients.
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Tejido Adiposo , Degeneración del Disco Intervertebral , Desplazamiento del Disco Intervertebral , Vértebras Lumbares , Músculos Paraespinales , Humanos , Músculos Paraespinales/diagnóstico por imagen , Músculos Paraespinales/patología , Desplazamiento del Disco Intervertebral/diagnóstico por imagen , Masculino , Femenino , Degeneración del Disco Intervertebral/diagnóstico por imagen , Degeneración del Disco Intervertebral/patología , Degeneración del Disco Intervertebral/etiología , Estudios Retrospectivos , Persona de Mediana Edad , Vértebras Lumbares/diagnóstico por imagen , Adulto , Tejido Adiposo/patología , Tejido Adiposo/diagnóstico por imagen , Tomografía Computarizada por Rayos X , Factores de Riesgo , AncianoRESUMEN
Presently, the clinical treatment of intervertebral disc degeneration (IVDD) remains challenging, but the strategy of simultaneously overcoming the overactive inflammation and restoring the anabolic/catabolic balance of the extracellular matrix (ECM) in the nucleus pulposus (NP) has become an effective way to alleviate IVDD. IL-1ra, a natural antagonist against IL-1ß, can mitigate inflammation and promote regeneration in IVDD. Chondroitin sulfate (CS), an important component of the NP, can promote ECM synthesis and delay IVDD. Thus, these were chosen and integrated into functionalized microspheres to achieve their synergistic effects. First, CS-functionalized microspheres (GelMA-CS) with porous microstructure, good monodispersion, and about 200 µm diameter were efficiently and productively fabricated using microfluidic technology. After lyophilization, the microspheres with good local injection and tissue retention served as the loading platform for IL-1ra and achieved sustained release. In in vitro experiments, the IL-1ra-loaded microspheres exhibited good cytocompatibility and efficacy in inhibiting the inflammatory response of NP cells induced by lipopolysaccharide (LPS) and promoting the secretion of ECM. In in vivo experiments, the microspheres showed good histocompatibility, and local, minimally invasive injection of the IL-1ra-loaded microspheres could reduce inflammation, maintain the height of the intervertebral disc (IVD) and the water content of NP close to about 70 % in the sham group, and retain the integrated IVD structure. In summary, the GelMA-CS microspheres served as an effective loading platform for IL-1ra, eliminated inflammation through the controlled release of IL-1ra, and promoted ECM synthesis via CS to delay IVDD, thereby providing a promising intervention strategy for IVDD. STATEMENT OF SIGNIFICANCE: The strategy of simultaneously overcoming the overactive inflammation and restoring the anabolic/catabolic balance of the extracellular matrix (ECM) in nucleus pulposus (NP) has shown great potential prospects for alleviating intervertebral disc degeneration (IVDD). From the perspective of clinical translation, this study developed chondroitin sulfate functionalized microspheres to act as the effective delivery platform of IL-1ra, a natural antagonist of interleukin-1ß. The IL-1ra loading microspheres (GelMA-CS-IL-1ra) showed good biocompatibility, good injection with tissue retention, and synergistic effects of inhibiting the inflammatory response induced by lipopolysaccharide and promoting the secretion of ECM in NPCs. In vivo, they also showed the beneficial effect of reducing the inflammatory response, maintaining the height of the intervertebral disc and the water content of the NP, and preserving the integrity of the intervertebral disc structure after only one injection. All demonstrated that the GelMA-CS-IL-1ra microspheres would have great promise for the minimally invasive treatment of IVDD.
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Sulfatos de Condroitina , Proteína Antagonista del Receptor de Interleucina 1 , Degeneración del Disco Intervertebral , Microesferas , Sulfatos de Condroitina/química , Sulfatos de Condroitina/farmacología , Degeneración del Disco Intervertebral/tratamiento farmacológico , Degeneración del Disco Intervertebral/patología , Animales , Proteína Antagonista del Receptor de Interleucina 1/farmacología , Conejos , Núcleo Pulposo/efectos de los fármacos , Núcleo Pulposo/patología , Núcleo Pulposo/metabolismo , Masculino , Matriz Extracelular/metabolismoRESUMEN
Intervertebral disc degeneration (IDD) is a highly prevalent musculoskeletal disorder that is associated with considerable morbidity. However, there is currently no drug available that has a definitive therapeutic effect on IDD. In this study, we aimed to identify the molecular features and potential therapeutic targets of IDD through a comprehensive multiomics profiling approach. By integrating transcriptomics, proteomics, and ultrastructural analyses, we discovered dysfunctions in various organelles, including mitochondria, the endoplasmic reticulum, the Golgi apparatus, and lysosomes. Metabolomics analysis revealed a reduction in total phosphatidylcholine (PC) content in IDD. Through integration of multiple omics techniques with disease phenotypes, a pivotal pathway regulated by the lysophosphatidylcholine acyltransferase 1 (LPCAT1)-PC axis was identified. LPCAT1 exhibited low expression levels and exhibited a positive correlation with PC content in IDD. Suppression of LPCAT1 resulted in inhibition of PC synthesis in nucleus pulposus cells, leading to a notable increase in nucleus pulposus cell senescence and damage to cellular organelles. Consequently, PC exhibits potential as a therapeutic agent, as it facilitates the repair of the biomembrane system and alleviates senescence in nucleus pulposus cells via reversal of downregulation of the LPCAT1-PC axis.
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1-Acilglicerofosfocolina O-Aciltransferasa , Degeneración del Disco Intervertebral , Fosfatidilcolinas , Humanos , Degeneración del Disco Intervertebral/metabolismo , Degeneración del Disco Intervertebral/tratamiento farmacológico , Degeneración del Disco Intervertebral/patología , Degeneración del Disco Intervertebral/genética , 1-Acilglicerofosfocolina O-Aciltransferasa/metabolismo , 1-Acilglicerofosfocolina O-Aciltransferasa/genética , Fosfatidilcolinas/metabolismo , Fosfatidilcolinas/química , Núcleo Pulposo/metabolismo , Núcleo Pulposo/patología , Metabolómica , Proteómica/métodos , Masculino , Senescencia Celular/efectos de los fármacos , Persona de Mediana Edad , Adulto , Femenino , Perfilación de la Expresión Génica , MultiómicaRESUMEN
Spinal magnetic resonance (MR) scans are a vital tool for diagnosing the cause of back pain for many diseases and conditions. However, interpreting clinically useful information from these scans can be challenging, time-consuming and hard to reproduce across different radiologists. In this paper, we alleviate these problems by introducing a multi-stage automated pipeline for analysing spinal MR scans. This pipeline first detects and labels vertebral bodies across several commonly used sequences (e.g. T1w, T2w and STIR) and fields of view (e.g. lumbar, cervical, whole spine). Using these detections it then performs automated diagnosis for several spinal disorders, including intervertebral disc degenerative changes in T1w and T2w lumbar scans, and spinal metastases, cord compression and vertebral fractures. To achieve this, we propose a new method of vertebrae detection and labelling, using vector fields to group together detected vertebral landmarks and a language-modelling inspired beam search to determine the corresponding levels of the detections. We also employ a new transformer-based architecture to perform radiological grading which incorporates context from multiple vertebrae and sequences, as a real radiologist would. The performance of each stage of the pipeline is tested in isolation on several clinical datasets, each consisting of 66 to 421 scans. The outputs are compared to manual annotations of expert radiologists, demonstrating accurate vertebrae detection across a range of scan parameters. Similarly, the model's grading predictions for various types of disc degeneration and detection of spinal metastases closely match those of an expert radiologist. To aid future research, our code and trained models are made publicly available.