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Rheumatoid arthritis (RA) is a chronic autoimmune disorder characterized by synovial inflammation and progressive joint destruction driven by macrophage polarization and osteoclast activation. Current therapies lack lesion-specificity and cause systemic side effects, highlighting the need for targeted treatment strategies. Here, we developed carrier-free myricetin-arginine conjugate nanozymes (MANZs) via a Mannich reaction-mediated conjugation of l-arginine and myricetin, followed by self-assembly driven by noncovalent interaction. The MANZs selectively target M1 macrophages via cationic amino acid transporter 2 (CAT2)-mediated uptake, facilitating preferential accumulation in inflamed joints. MANZs exert multi-modal therapeutic effects by scavenging reactive oxygen species (ROS), repolarizing M1 macrophages, and inhibiting osteoclast differentiation. In a collagen-induced arthritis (CIA) mouse model, MANZs significantly alleviated joint swelling, synovitis, and bone erosion without systemic toxicity. This work establishes a promising paradigm for RA therapy by integrating cationic amino acids with natural polyphenols into a self-assembled, target-specific nanoplatform with high biocompatibility and translational potential.
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BACKGROUND: MicroRNAs (miRNAs) are crucial regulators of osteoclast differentiation. Icariin (ICA), the primary active constituent of the traditional Chinese herb Epimedium, has been shown to inhibit osteoclastogenesis. However, whether ICA suppresses osteoclast differentiation via miRNA regulation remains to be elucidated. METHODS: We employed bioinformatic analysis coupled with in vivo validation to screen for and identify potential targets that mediate the effects of ICA on osteoclast formation. Subsequently, we carried out in vitro experiments involving the overexpression and knockdown of miR-376c-3p in bone marrow-derived macrophages (BMMs). Additionally, luciferase reporter assays and western blot analysis were utilized to ascertain the direct targets of miR-376c-3p. RESULTS: It was confirmed that miR-376c-3p plays a regulatory role in osteoclast differentiation within bone marrow-derived macrophages (BMMs). Notably, the knockdown of miR-376c-3p led to a partial reversal of the inhibitory effect that ICA exerts on osteoclastogenesis. Furthermore, matrix metalloproteinase-3 (Mmp3) was validated as a direct target of miR-376c-3p, providing evidence to suggest that ICA likely mediates its effects through the miR-376c-3p/Mmp3 signaling axis. CONCLUSION: Our findings demonstrate that ICA exerts its anti-osteoporotic effect by downregulating the miR-376c-3p-mediated Mmp3 expression. This study provides insights that potentially facilitate ICA's clinical application.
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This review focuses on experimental models developed to study myeloma bone disease (MBD), a major cause of morbidity in multiple myeloma (MM). Under physiological conditions, bone remodeling is regulated by osteoclasts (OCs) and osteoblasts (OBs); in MM, this balance is disrupted, resulting in enhanced bone resorption and suppressed bone formation. Myeloma cells alter the bone marrow (BM) microenvironment by increasing the RANKL/OPG ratio and secreting Wnt pathway inhibitors such as DKK-1 and sclerostin, thereby promoting osteoclastogenesis and inhibiting osteoblast differentiation. To dissect these mechanisms and evaluate therapeutic strategies, diverse preclinical systems have been developed. Syngeneic murine models, notably the 5T series, remain the most established for reproducing both osteolysis and impaired bone formation, though interspecies differences limit translational relevance. Humanized mouse systems and three-dimensional (3D) in vitro models increasingly address these constraints by incorporating human stromal and hematopoietic elements. Emerging induced pluripotent stem cell-derived bone marrow organoids (iBMOs) offer a fully human platform capable of modeling both osteoclast and osteoblast dynamics. While current iBMOs lack mineralized bone and mature vascular or immune components, advances in differentiation control and matrix engineering are expected to bridge these gaps, providing physiologically relevant and ethically sustainable models for studying MBD and testing therapeutic interventions.
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Recent studies support that hematopoietic stem cell (HSC)-derived myeloid dendritic cells, monocytes/macrophages (Mo/MÏ), and osteoclast precursors (OCps) share common progenitor(s) during development. This occurs mainly through receptor activator of NF-κB ligand (RANKL) signaling via its cytoplasmic adaptor protein complex (TRAF6) to subsequent osteoclastogenesis for bone loss and/or remodeling. Presently, mounting new evidence suggests that erythro-myeloid progenitor (EMP)-derived macrophages (MÏ) and HSC-derived monocytes (Mo) produce embryonic, fetal, and postnatal OCp pools (i.e., primitive OCp), pinpointing a complex network of multiple OCp developmental origins. However, their ontogenic developments, lineage interactions, and contributions to the alternative osteoclastogenesis-in contrast to overall bone remodeling or loss-remain elusive. Interestingly, studies have also elucidated the contributions of immature CD11c+ myeloid DC-like OCps to osteoclastogenesis, with or without the classical so-called Mo/MÏ-derived OCp subsets, and described that CD11c+ myeloid DCs (mDCs) develop into functionally active OCs; meanwhile, the cytokine TGF-ß mediates a stepwise regulation of de novo immature mDCs/OCps through distinct crosstalk(s) with IL-17, an unrecognized interaction featuring TRAF6(-/-)CD11c+ mDDOCps that coexist and proficiently colocalize in the local environment to drive a bona fide route for alternative osteoclastogenesis in vivo. Collectively, new findings-critically hinged on progenitor osteoclastogenic pathways (primitive OCps, mDCs/OCps, osteomorphs, etc.) and involving classical and/or alternative routes to inflammation-induced bone loss-are discussed via the illustrated schemes. This review highlights plausible ontogenic vs. principal or alternative developmental paths and their consequential downstream effects.
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Antígeno CD11c , Células Dendríticas , Osteoclastos , Osteogénesis , Humanos , Animales , Osteoclastos/citología , Osteoclastos/metabolismo , Células Dendríticas/metabolismo , Células Dendríticas/citología , Diferenciación Celular , Transducción de Señal , Células Mieloides/metabolismo , Células Mieloides/citología , Macrófagos/metabolismo , Macrófagos/citología , Antígeno CD11c/metabolismoRESUMEN
Wear particle-induced periprosthetic osteolysis (PPO) represents the primary reason for implant failure following joint replacement, driven by macrophage polarization and excessive osteoclastogenesis. While cell migration inducing hyaluronidase 1 (CEMIP) has been implicated in inflammation and bone metabolism, its role in wear particle-induced PPO remains unexplored. Here, we discover that CEMIP expression was upregulated in Ti particle-induced calvarial osteolysis model. Functional studies revealed that CEMIP knockdown could attenuate Ti particle-triggered bone resorption via suppressing macrophage M1 polarization, promoting M2 polarization, and inhibiting osteoclast differentiation, as evidenced by reduced M1 markers, increased M2 markers, decreased inflammatory cytokines, and osteoclast-specific gene expression. In vitro, CEMIP silencing suppressed IFNγ-induced M1 polarization, enhanced IL-4-induced M2 polarization, and RANKL/M-CSF-triggered osteoclastogenesis in bone marrow-derived macrophages (BMDMs). Mechanistically, wear particle exposure decreases N6-methyladenosine (m6A) modification of CEMIP mRNA, impairing m6A reader protein YTH N6-methyladenosine RNA binding protein F2 (YTHDF2)-mediated recognition and subsequent mRNA decay, thereby stabilizing CEMIP transcripts and enhancing their expression. Furthermore, CEMIP knockdown counteracted the enhancement of M1 macrophage polarization and osteoclast differentiation resulting from YTHDF2 silencing. Collectively, these findings highlight the critical role of the YTHDF2/CEMIP axis in regulating macrophage polarization and osteoclast differentiation, thus offering a promising therapeutic target for wear particle-induced PPO.
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Macrófagos , Osteólisis , Proteínas de Unión al ARN , Animales , Osteólisis/metabolismo , Osteólisis/inducido químicamente , Ratones , Proteínas de Unión al ARN/metabolismo , Proteínas de Unión al ARN/genética , Ratones Endogámicos C57BL , Macrófagos/inmunología , Osteoclastos , Titanio , Adenosina/análogos & derivados , Adenosina/metabolismo , Masculino , Osteogénesis , Humanos , Diferenciación CelularRESUMEN
Renal osteodystrophy (ROD) is a major debilitating skeletal complication of chronic kidney disease characterized by dysregulated bone remodeling. Neutrophils are essential for bone homeostasis. However, their functionality in the ROD osteoimmune microenvironment remains unclear. To investigate the crosstalk between neutrophils and bone metabolic imbalance in ROD, we firstly performed transcriptomic profiling in patients with ROD, revealing a pronounced disruption of bone homeostasis. These findings were corroborated in a murine model of ROD, which demonstrated an osteoporotic phenotype and increased osteoclast activity. Through integrated bioinformatics and experimental validation, we identified substantial functional impairments of neutrophils in the ROD bone microenvironment, including downregulated degranulation, reduced neutrophil extracellular trap formation, and decreased abundance. In co-culture systems, ROD-derived neutrophils potently enhanced osteoclastogenesis from bone marrow-derived macrophages indirectly through the modulation of cytokine release, which was mediated by the PI3K-Akt signaling pathway. Furthermore, potential biomarkers and a therapeutic compound were identified and preliminarily validated via in silico and in vitro experiments, highlighting the translational prospects for ROD management. Our findings demonstrated that neutrophil dysfunction disrupts osteoclastogenesis-osteogenesis equilibrium, contributing to pathological bone loss in the ROD bone microenvironment.
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Resorción Ósea , Trastorno Mineral y Óseo Asociado a la Enfermedad Renal Crónica , Neutrófilos , Osteoclastos , Osteogénesis , Animales , Neutrófilos/inmunología , Trastorno Mineral y Óseo Asociado a la Enfermedad Renal Crónica/inmunología , Trastorno Mineral y Óseo Asociado a la Enfermedad Renal Crónica/genética , Trastorno Mineral y Óseo Asociado a la Enfermedad Renal Crónica/patología , Trastorno Mineral y Óseo Asociado a la Enfermedad Renal Crónica/metabolismo , Ratones , Biología Computacional , Humanos , Osteogénesis/inmunología , Masculino , Ratones Endogámicos C57BL , Modelos Animales de Enfermedad , Femenino , Transducción de Señal , Resorción Ósea/inmunología , MacrófagosRESUMEN
INTRODUCTION: Root resorption is a pathologic process characterized by the breakdown of dentin and cementum by odontoclasts, mirroring the mechanisms of osteoclast-driven bone resorption. Osteoclastogenesis is tightly regulated by the RANK/RANKL/OPG axis and other signaling pathways, including Wnt, ATP-P2RX7-IL-1, programmed cell death, and inflammasome activation. OBJECTIVE: This review provides a comprehensive analysis of the key signaling pathways and molecular mediators orchestrating root resorption in orthodontic, traumatic, and inflammatory conditions. METHODS: A literature-based analysis was conducted, focusing on molecular and cellular mechanisms involved in root resorption. Key pathways such as RANK/RANKL/OPG, Wnt signaling, ATP-P2RX7-IL-1, and inflammasome activation were examined. The role of proinflammatory and anti-inflammatory cytokines, matrix metalloproteinases, and periostin were also analyzed. RESULTS: Proinflammatory mediators such as IL-1, IL-6, IL-8, tumor necrosis factor-alpha, IL-17, IL-22, and IL-23 drive odontoclastic differentiation, whereas anti-inflammatory cytokines, including IL-4, IL-10, and transforming growth factor-beta, counteract resorptive activity. Additionally, matrix metalloproteinases and periostin modulate extracellular matrix remodeling, impacting resorption progression. The balance between resorptive and reparative processes is influenced by the inflammatory microenvironment, fibroblast-macrophage interactions, and mechanotransduction. While the molecular mechanisms underlying odontoclastogenesis parallel bone resorption, unique features of root structures, such as the cementoid layer, contribute to resistance against resorption. CONCLUSION: The intricate cross talk between pro-resorptive and antiresorptive factors, emphasizing their roles in odontoclast activation and extracellular matrix remodeling, dictate the extent of root degradation.
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Inflamación , Osteoclastos , Resorción Radicular , Transducción de Señal , Humanos , Resorción Radicular/metabolismo , Resorción Radicular/fisiopatología , Osteoclastos/fisiología , Osteoclastos/metabolismo , Transducción de Señal/fisiología , Inflamación/fisiopatología , Inflamación/metabolismo , Citocinas/metabolismo , Animales , Osteogénesis/fisiología , Remodelación Ósea/fisiologíaRESUMEN
Osteoporosis (OP) is a disease characterized by decreased bone mass and damaged architectures. The promising treatment strategy for OP is to inhibit bone resorption while promoting bone formation. MicroRNAs (miRNAs) have been shown to be associated with osteoclastogenesis and osteogenesis processes in OP. In our previous study, we discovered that miR-378 inhibits bone marrow mesenchymal stem cell (BMSC) osteogenesis and bone formation during fracture healing. However, the role of miR-378 during OP progression is not validated. In this study, we found that miR-378 transgenic (Tg) mice exhibited excessive bone loss after ovariectomy (OVX) treatment. MiR-378 increased BMSC's osteoclastogenesis by activating both canonical and non-canonical nuclear factor kappa-light-chain-enhancer of activated B (NFκB) signaling pathway. Tumor necrosis factor receptor-associated factor 3 (Traf3) was directly regulated by miR-378 during osteoclast differentiation. miR-378 also aggravated transforming growth factor beta (TGFß) impaired osteogenesis upon OVX treatment. Traf3 was involved in this process as well. In in vivo study, the intravenous injection of anti-miR-378 lentivirus could significantly rescue OVX induced bone loss and bone microarchitecture impairment. This study uncovered the novel role of miR-378 in OVX induced osteoporosis. The potential of developing miRNA-378 inhibitors as novel diagnostics or blockers as therapeutics for osteoporosis is worth exploring.
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Osteoclast functioning determines homeostasis of bone texture, healing speed after bone injury, and integrity after bone remodeling. Osteoclastogenesis is tightly controlled by transcriptional factors and cytokine-mediated signaling. The activating transcription factor 4 (Atf4) and interleukin 6 (IL6) have been considered to be involved in osteoclastogenesis, but their interaction on osteoclast differentiation is still largely unclear. Here we aimed to investigate their interaction on osteoclast differentiation and activity. Through lentiviral overexpression and siRNA transfection, we showed that Atf4 regulated osteoclastogenesis by upregulating the expression of osteoclast markers, including Nfatc1, c-Fos, Dcst1, Mmp2 and 9, and cathepsin K. Atf4-mediated osteoclastogenesis was largely dependent on activation of IL6 signaling through the direct binding of Atf4 to the IL6 promoter by ChIP assay. Atf4-IL6 axis activated the Jak1/Stat3 signaling to promote the formation of large, multinucleated osteoclasts. Taken together, these results indicate the important interaction between Atf4 and IL6 signaling on osteoclast differentiation beyond canonical MCSF and RANKL signaling and emphasize the multi-regulatory mechanism of Atf4-mediated osteoclastogenesis, which provides us with understanding of osteoporotic diseases and cues for therapeutic strategies.
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Factor de Transcripción Activador 4 , Diferenciación Celular , Interleucina-6 , Osteoclastos , Osteogénesis , Osteoclastos/metabolismo , Osteoclastos/citología , Factor de Transcripción Activador 4/metabolismo , Factor de Transcripción Activador 4/genética , Animales , Diferenciación Celular/fisiología , Ratones , Interleucina-6/metabolismo , Interleucina-6/genética , Transducción de Señal/fisiología , Osteogénesis/fisiología , Ratones Endogámicos C57BL , Factores de Transcripción NFATC/metabolismo , Células RAW 264.7 , Factor de Transcripción STAT3/metabolismo , Ligando RANK/metabolismo , Células CultivadasRESUMEN
Osteoporosis is a chronic bone disease characterized by impaired bone remodeling and increased fracture risk. While classical mechanisms implicate estrogen deficiency, aging, and altered receptor activator of the nuclear factor-κB ligand (RANKL)/osteoprotegerin (OPG) signaling, growing evidence supports a pivotal role of immune and inflammatory pathways in sustaining osteoclast-mediated bone resorption. A distinctive hallmark observed in osteoporotic patients is spontaneous osteoclastogenesis (SO), defined as the ability of mononuclear precursors to differentiate into osteoclasts even in the absence of exogenous stimuli such as RANKL or macrophage colony-stimulating factor (M-CSF), a process driven by an intrinsically primed in vivo microenvironment that includes platelets. We hypothesize that platelets may contribute to this priming not only through soluble mediators but also via the release of extracellular vesicles, particularly exosomes enriched in regulatory microRNAs (miRs). Within this framework, platelet-derived exosomal miRs (P-EXO-miRs) may orchestrate multiple intercellular interactions within the bone marrow microenvironment, modulating monocytes, macrophages, stromal and endothelial cells, as well as T and B lymphocytes. Specifically, miR-21, miR-223, miR-214, and miR-155 emerge as key candidates capable of regulating cytokine secretion, inflammatory signaling, and the RANKL/OPG balance, thereby promoting a pro-osteoclastogenic milieu. Network-based analysis using miRNet further supports the involvement of these miRs in pathways such as Hedgehog, Wnt, and actin cytoskeleton regulation, all relevant to osteoclast differentiation and function. Through these mechanisms, P-EXO-miRs may amplify chronic low-grade inflammation and facilitate spontaneous osteoclast differentiation and activity, ultimately contributing to bone loss in osteoporosis. Future investigations should aim to experimentally validate this platelet-bone axis, delineate the molecular targets of individual miRs, and explore their potential as circulating biomarkers or therapeutic targets. By unveiling this previously unrecognized role of platelet-derived miRs in SO, this hypothesis opens new perspectives for the understanding, early detection, and treatment of osteoporosis.
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Bruceine A (BA) is a quassinoid compound derived from Brucea javanica with various pharmacological activities. Our previous work has revealed that BA exhibits anti-inflammatory and glucose-lowering effects in db/db mice, resulting in significant protection against diabetic kidney disease. However, considering the pivotal role of inflammatory signaling pathways during osteoclast differentiation, whether and how BA exerts protection against osteoporosis remains to be explored. In the present work, we have demonstrated that BA exhibits significant protective efficacy against bone loss in ovariectomized and diabetic mice, which mimic postmenopausal and diabetic osteoporosis, respectively. Moreover, BA is found to attenuate RANKL-induced osteoclast differentiation and function in isolated bone marrow-derived macrophages and the RAW264.7 cell line at non-cytotoxic concentrations. Mechanistically, BA binds to RACK1 to disrupt the RACK1-c-SRC interaction, thereby suppressing the inflammatory signaling pathways involved in regulating osteoclastogenesis in a RACK1-dependent manner. Altogether, our findings establish BA as a novel therapeutic agent for treating osteoporosis.
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Osteoclastos , Osteogénesis , Osteoporosis , Cuassinas , Receptores de Cinasa C Activada , Animales , Receptores de Cinasa C Activada/metabolismo , Ratones , Osteoporosis/tratamiento farmacológico , Osteoporosis/metabolismo , Osteoporosis/patología , Osteoclastos/efectos de los fármacos , Osteoclastos/metabolismo , Osteogénesis/efectos de los fármacos , Femenino , Células RAW 264.7 , Ratones Endogámicos C57BL , Cuassinas/farmacología , Diferenciación Celular/efectos de los fármacos , Transducción de Señal/efectos de los fármacos , Macrófagos/metabolismo , Macrófagos/efectos de los fármacosRESUMEN
Metastasis in breast cancer frequently spreads to the bones, significantly impacting patient outcomes and escalating mortality rates. The ataxia-telangiectasia mutated (ATM) kinase plays a pivotal role in regulating the DNA damage response (DDR) and has been linked to the invasion and spread of breast cancer. In this study we investigated the regulatory mechanisms of ATM in bone metastasis of breast cancer. The bone metastases models were constructed in female nude mice: The MDA-MB-231 tumor model was generated by implanting luciferase-tagged MDA-MB-231 cells into the left hind tibia and intra-caudal artery. For the SK-BR-3 tumor model, luciferase-tagged SK-BR-3 cells were injected through the intra-caudal artery. By conducting bioinformatics analyses and in vitro and in vivo experiments, we found that ATM expression was markedly elevated in bone metastasis samples compared to liver, lung or skin metastases. We demonstrated that ATM boosted the migrative and invasive abilities and pre-osteoclast differentiation of MDA-MB-231 and SK-BR-3 cell lines via expression of CCL2, an osteoclast-related cytokine. The regulation of ATM on CCL2 was found to be NFκB dependent. In vivo experiments confirmed that ATM knockout (ATM KO) or treatment with small-molecule ATM inhibitor KU55933 markedly inhibited osteoclastogenesis of SK-BR-3 cells and the progression of breast cancer bone metastasis. Our results underscore the pivotal role of ATM in regulating NFκB-CCL2 expression and promoting the progression of breast cancer bone metastasis.
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Bone-related disorders (BRDs), such as osteoporosis, rheumatoid arthritis, and osteosarcoma, are major contributors to morbidity and disability worldwide. Conventional treatments are often limited by poor targeting, systemic toxicity, and insufficient long-term efficacy. Selenium (Se), an essential trace element, plays a pivotal role in maintaining redox homeostasis, immune balance, and bone remodeling. Selenium-containing nanoparticles (SeNPs) have emerged as promising platforms that integrate the biological activities of selenium with the tunable features of nanomaterials. This review provides a comprehensive overview of SeNPs, covering synthesis strategies, physicochemical properties, and their roles in regulating osteogenesis, osteoclastogenesis, oxidative stress, and inflammatory signaling in the skeletal microenvironment. We further highlight recent advances in applying SeNPs for the treatment of BRDs, including their incorporation into biomaterials and combination therapies such as photothermal and chemodynamic approaches. While preclinical studies show encouraging results, challenges remain in understanding long-term biosafety, biodistribution, and clinical translation. Overall, SeNPs-based nanomedicine offers significant potential for precision bone-targeted therapies and tissue regeneration.
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Enfermedades Óseas , Huesos , Nanopartículas , Selenio , Humanos , Selenio/administración & dosificación , Selenio/uso terapéutico , Selenio/química , Nanopartículas/química , Nanopartículas/uso terapéutico , Nanopartículas/administración & dosificación , Animales , Enfermedades Óseas/tratamiento farmacológico , Enfermedades Óseas/metabolismo , Huesos/metabolismo , Huesos/efectos de los fármacos , Osteogénesis/efectos de los fármacosRESUMEN
Bone homeostasis depends on spatially orchestrated interactions among osteoclasts, osteoblasts, and osteocytes that are embedded within a unique extracellular matrix that is mineralized on the nanoscale to define the structure and function of bone. Reconstructing these interactions to enable autonomous cell differentiation and tissue remodeling has remained a significant challenge towards mimicking adequate bone physiology in-vitro. Here, we present an engineered model that spatially defines the paracrine communication of heterogeneous cell populations within bone tissue that support the rapid maturation of primary osteoblasts into osteocytes, the differentiation of macrophages into osteoclasts, and calcified tissue resorption within a mineralized cell-laden bone-like tissue. We demonstrate that nanoscale mineralization of cell-laden collagen hydrogels on-a-chip enhances osteoblast to osteocyte differentiation, whereas osteocytes in the matrix accelerate osteoclastogenesis and remodeling in a spatially defined manner without the need for exogenous growth factors. Osteocyte-dependent osteoclastogenesis on-a-chip outperformed conventional stimulation with RANKL and M-CSF, reproduced the clinical response of anti-resorptive drugs, and mimicked established tumor-bone interactions observed in invasive oral cancer. By replicating essential aspects of bone composition and function, this system provides a robust, self-regulated microphysiologic model to investigate bone remodeling, cancer-bone crosstalk, and therapeutic interventions.
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Osteoporosis, characterized by excessive osteoclast-mediated bone resorption, necessitates innovative therapies to overcome the limitations of current treatments. This study explores the effects of epifriedelinol (EFD), a triterpene obtained from Aster tataricus L.fil., on RANKL-induced osteoclastogenesis in RAW264.7 cells and bone marrow-derived macrophages. Concentrations up to 10 µM of EFD exhibited no cytotoxicity but strongly impeded RANKL-induced osteoclast differentiation, as indicated by a decrease in TRAP-positive multinucleated cells in both cell types. EFD at least partially inhibited the activation of the MAPK signaling pathways (ERK, JNK, p38) and suppressed key transcription factors NFATc1 and c-Fos, essential for osteoclast-specific gene expression (TRAP, RANK, MMP-9, cathepsin K). Moreover, EFD reduced RANKL-induced bone resorption in BMMs, demonstrating decreased resorption pit area in a dose-dependent manner. These findings indicate that EFD inhibits osteoclast differentiation and function by targeting the MAPK-mediated NFATc1/c-Fos pathway, highlighting its potential as a natural treatment for osteoclast-related disorders such as osteoporosis.
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Resorción Ósea , Diferenciación Celular , Factores de Transcripción NFATC , Osteoclastos , Proteínas Proto-Oncogénicas c-fos , Triterpenos , Animales , Ratones , Factores de Transcripción NFATC/metabolismo , Factores de Transcripción NFATC/antagonistas & inhibidores , Osteoclastos/efectos de los fármacos , Osteoclastos/citología , Osteoclastos/metabolismo , Diferenciación Celular/efectos de los fármacos , Proteínas Proto-Oncogénicas c-fos/metabolismo , Proteínas Proto-Oncogénicas c-fos/antagonistas & inhibidores , Células RAW 264.7 , Resorción Ósea/tratamiento farmacológico , Resorción Ósea/metabolismo , Resorción Ósea/patología , Ligando RANK/farmacología , Ligando RANK/metabolismo , Macrófagos/efectos de los fármacos , Macrófagos/metabolismo , Macrófagos/citología , Relación Dosis-Respuesta a Droga , Proteínas Quinasas Activadas por Mitógenos/metabolismo , Proteínas Quinasas Activadas por Mitógenos/antagonistas & inhibidores , Triterpenos/farmacología , Triterpenos/química , Triterpenos/aislamiento & purificación , Relación Estructura-Actividad , Sistema de Señalización de MAP Quinasas/efectos de los fármacos , Supervivencia Celular/efectos de los fármacosRESUMEN
The development of craniofacial bones and teeth relies heavily on the Wnt signaling pathway, yet the specific mechanisms and Wnt variants involved remain under continual investigation. Using publicly available single-cell sequencing data from the mouse incisor, we reveal Wnt1 expression across dental structures and investigate its role using a Col1a1-dependent Wnt1 transgenic mouse model. Inducing Wnt1 early on affects craniofacial bone without disturbing tooth development, but prolonged embryonic induction leads to postnatal mortality with osteopetrosis-like bone overgrowth and malformed teeth. While tooth formation was initially unaffected by postnatal Wnt1 induction, prolonged activation impaired tooth root formation and odontoblast differentiation, resulting in shortened roots and thinner dentin. Three-dimensional micro-computed tomography quantification reveal that both embryonic and postnatal activation of Wnt1 significantly increase neural crest-derived craniofacial bone volume, whereas mesenchymal-derived craniofacial bones are unaffected. Importantly, osteoclastogenesis is suppressed by Wnt1 in a dose-dependent manner, revealed through bulk RNA sequencing and in vitro experiments. These findings emphasize the differential effects of Wnt1 on bone development based on origin and highlight its role in modulating osteoclast activity, indicating broader implications for craniofacial development and potential therapeutic avenues.
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Huesos Faciales , Odontogénesis , Cráneo , Proteína Wnt1 , Animales , Proteína Wnt1/fisiología , Proteína Wnt1/genética , Proteína Wnt1/metabolismo , Ratones , Ratones Transgénicos , Odontogénesis/genética , Odontogénesis/fisiología , Microtomografía por Rayos X , Huesos Faciales/embriología , Huesos Faciales/crecimiento & desarrollo , Diferenciación Celular , Vía de Señalización Wnt , Odontoblastos , Osteogénesis , Cresta Neural , Cadena alfa 1 del Colágeno Tipo IRESUMEN
Orthodontic treatment depends on periodontal tissue remodeling. While the immune system has been found to regulate this process, previous studies mostly focused on the static molecular changes in conventional immune cell sets. A recent study used single-cell RNA sequencing (scRNA-seq) to analyze macrophages under orthodontic tooth movement (OTM), introducing new subclusters and functions. However, changes in other immune cells and their interactions with surrounding stromal cells are yet unclarified. Therefore, we performed scRNA-seq in mice, aiming to describe a more comprehensive immune landscape during OTM, while further exploring the dynamic changes, functions, and cellular interactions of immune cells at a higher resolution. We first confirmed the dynamic activation of osteoclasts, osteoblasts, and immune cells at various time points. The scRNA-seq analysis identified 7 cell lineages and 18 major cell types, with immune cells forming the largest proportion. Monocytic cells, granulocytes, and lymphocytes were selected for individual reclustering, followed by analysis of specific gene expression, functional enrichment, and distribution changes during OTM. Pseudotime analysis was applied to monocytic cells and granulocytes. We identified 4 developmental pathways in monocytic cells toward dendritic cells, different subsets of macrophages, and osteoclasts. Monocytic cells tended to be more differentiated during OTM. Meanwhile, in granulocytes, neutrophil subclusters were all highly differentiated. Additionally, we assessed the cellular interactions during OTM, revealing enhanced signaling from macrophages toward osteoclasts, especially in Ccl, Tnf, and Spp1 pathways. We identified that the C3ar1+ macrophage subcluster expressed these cytokines at a high level and was enriched in positive regulation of the mitogen-activated protein kinase cascade, indicating its positive regulation toward osteoclast activity. In conclusion, this study revealed the complex immune microenvironment during OTM, providing a detailed perspective on the diverse immune cell types, their specific functions, and cellular interactions.
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Microambiente Celular , Análisis de la Célula Individual , Técnicas de Movimiento Dental , Animales , Ratones , Osteoclastos/inmunología , Macrófagos/inmunología , Microambiente Celular/inmunología , Ratones Endogámicos C57BL , Análisis de Secuencia de ARN , Osteoblastos/inmunología , Monocitos/inmunologíaRESUMEN
Two new chromone derivatives, cnidimols I and J (1 and 2), together with ten known aromatic derivatives (3-12), were isolated from the Beibu Gulf algicolous fungus Aspergillus versicolor GXIMD 02518. Their structures were determined by comprehensive physicochemical and spectroscopic data interpretation. The absolute configurations of 1 and 2 were accomplished by ECD calculations and X-ray diffraction analysis. Compound 1 was obtained as a pair of enantiomers, which were separated by chiral-phase HPLC analysis. Notably, 3,7-dihydroxy-1,9-dimethyldibenzofuran (6) displayed significant inhibition in LPS-induced NF-κB luciferase activity in RAW 264.7 macrophages, which further inhibited RANKL-induced osteoclast differentiation without cytotoxicity in bone marrow macrophage cells.
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Aspergillus , Cromonas , Osteoclastos , Osteogénesis , Animales , Aspergillus/química , Ratones , Células RAW 264.7 , Osteoclastos/efectos de los fármacos , Osteoclastos/metabolismo , Cromonas/farmacología , Cromonas/química , Cromonas/aislamiento & purificación , Diferenciación Celular/efectos de los fármacos , Osteogénesis/efectos de los fármacos , Macrófagos/efectos de los fármacos , Macrófagos/metabolismo , FN-kappa B/metabolismo , Ligando RANK , Organismos Acuáticos , Estructura Molecular , Lipopolisacáridos/farmacologíaRESUMEN
This study identified six novel azaphilones, isochromophilones G-L (1-6), and three novel biosynthetically related congeners (7-9) from Diaporthe sp. SCSIO 41011. The structures and absolute configurations were elucidated through comprehensive spectroscopic analyses combined with experimental and calculated electronic circular dichroism (ECD) spectra. Significantly, three highly oxygenated azaphilones contain an acetyl group at the terminal chain (4) or linear conjugated polyenoid moieties (5 and 6), which occur infrequently in the azaphilone family. Additionally, several compounds demonstrated inhibition of lipopolysaccharide (LPS)-induced nuclear factor kappa-B (NF-κB) activation in RAW 264.7 macrophages at 20 µmol·L-1. The novel compound (1) effectively inhibited receptor activator of NF-κB ligand (RANKL)-induced osteoclast differentiation without exhibiting cytotoxicity in bone marrow and RAW 264.7 macrophages, indicating its potential as a promising lead compound for osteolytic disease treatment. This research presents the first documented evidence of azaphilone derivatives as inhibitors of RANKL-induced osteoclastogenesis.
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Ascomicetos , Benzopiranos , Osteoclastos , Osteogénesis , Pigmentos Biológicos , Ligando RANK , Animales , Ratones , Ligando RANK/metabolismo , Ligando RANK/genética , Células RAW 264.7 , Osteoclastos/efectos de los fármacos , Osteoclastos/citología , Osteoclastos/metabolismo , Benzopiranos/química , Benzopiranos/farmacología , Benzopiranos/aislamiento & purificación , Osteogénesis/efectos de los fármacos , Macrófagos/efectos de los fármacos , Macrófagos/citología , Macrófagos/metabolismo , Estructura Molecular , Pigmentos Biológicos/química , Pigmentos Biológicos/farmacología , Pigmentos Biológicos/aislamiento & purificación , Ascomicetos/química , FN-kappa B/metabolismo , FN-kappa B/genética , Diferenciación Celular/efectos de los fármacosRESUMEN
Background: Bone metastasis of breast cancer (BC) is a key reason for poor prognosis. Recently, natural ingredients derived from plants have been found to exert a broad anti-tumor effect and are considered to be promising candidates for adjuvant therapy. Astragalin (AS) was found to inhibit the progression of several types of tumors; however, the role of AS in regulating the bone metastasis of BC is still unclear. Methods: The effects of AS on the progression of bone metastasis of BC were detected in vivo through safranin O and fast green staining, in vivo living imaging and microCT. The BrdU assay and Annexin V-PI analysis were used to detect the effects of AS on the growth of BC cells. Furthermore, TRAP staining was performed to examine the formation of osteoclasts regulated by AS. A transcriptome was performed to explore the downstream effects of AS on regulating the growth of BC cells, and the mechanism was further confirmed by Western blot and real-time PCR. Results: Administration of AS could effectively attenuate the bone destruction and the progression of bone metastasis of BC. The growth of BC cells can be inhibited by AS by inducing ER stress-mediated upregulation of Ddit3. In addition, AS can also prevent osteoclastogenesis through inhibiting the activation of the AKT pathway. Conclusions: Our studies suggest that AS could be an ideal adjuvant therapy for attenuating the progression of bone metastasis of BC, since it can directly restrict the growth of tumor, as well as attenuate osteolysis.