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BACKGROUND: Postmenopausal osteoporosis significantly increases the risk of fracture, which seriously affects the quality of life of patients. Exercise therapy is an important non-drug means and prevention and treatment strategy for patients with osteoporosis, in which strength training is the best mode, but its specific biological mechanism has not been determined. OBJECTIVE: To investigate the effects of strength training on bone morphology, materials and biomechanics in ovariectomized rats and to explore the mechanism of extracellular matrix remodeling. METHODS: Forty-eight female Sprague-Dawley rats were divided into sham operation group, sham operation exercise group, ovariectomized group and ovariectomized exercise group according to the random number table method. The menopausal animal model was established by bilateral ovariectomy in the ovariectomized group and ovariectomized exercise group, while sham operation was performed in the sham operation group and sham operation exercise group. Four weeks after operation, the sham operation exercise group and the ovariectomized exercise group underwent 12-week tail weight-bearing ladder training, and the sham operation group and the ovariectomized group were raised quietly in the cage. The bilateral femur and tibia were separated after training. The right tibia was used for dual-energy X-ray densitometry and biomechanical, biophysical and biochemical analyses, the left tibia was detected using micro-computed tomography for bone microstructural examination, the right femur was subjected to hematoxylin-eosin staining for histological observation, and the left femur was used for western blot and gelatin zymography detection of protein expression and enzyme activity of extracellular matrix metabolismrelated factors, respectively. RESULTS AND CONCLUSION: Compared with the sham operation group, the maximal load and stiffness decreased (P < 0.05), bone density, bone mineral density, bone inorganic matter content, bone calcium content decreased (P < 0.05), bone water content increased (P < 0.05), trabecular bone volume fraction, trabecular connectivity density, and trabecular number decreased (P < 0.05), trabecular separation, structural model index increased (P < 0.05), bone adipocyte number and cross-sectional area increased (P < 0.05), matrix metalloproteinase-2 activity decreased (P < 0.05), and protein expression of tissue inhibitor of metalloproteinase-1 and osteoprotegerin increased (P < 0.05) in the ovariectomized group. Compared with the ovariectomized group, the maximal load, stiffness, fracture load and resilience increased (P < 0.05), bone mineral density, bone mineral content, bone mineral density, bone inorganic matter content, and bone calcium content increased (P < 0.05), bone water content decreased (P < 0.05), trabecular separation and bone marrow area decreased (P < 0.05), trabecular bone thickness, cortical bone volume fraction, cortical bone area fraction, cortical bone thickness, and cortical bone porosity increased (P < 0.05), bone adipocyte number and cross-sectional area reduced (P < 0.05), matrix metalloproteinase-2 activity increased (P < 0.05), and protein expression of tissue inhibitor of metalloproteinase-1, Runt-related transcription factor 2 and osteoprotegerin decreased (P < 0.05) in the ovariectomized exercise group. To conclude, strength training can protect against bone injury caused by estrogen deficiency, which is characterized by improvement of bone biomechanical properties, bone tissue composition and bone microstructure, and its mechanism is related to the regulation of extracellular matrix remodeling. [ABSTRACT FROM AUTHOR]

Background: Multiple epiphyseal dysplasia-4 (MED-4, MIM 226900) is a rare autosomal recessive disease characterized by disproportionate height and early onset osteoarthritis of the lower limbs. MED-4 is caused by homozygous or compound heterozygous pathogenic variants in the SLC26A2 gene. However, the underlying pathogenic mechanisms in chondrocytes remains unknown. This study aimed to identify the pathogenic variants within a MED-4 family and explore the molecular etiology of this condition in human primary chondrocyte cells. Methods: Clinical data were recorded and peripheral blood samples were collected for analysis. Whole exome sequencing (WES) and bioinformatic analyses were performed to determine causative variants. Wild-type SLC26A2 and corresponding mutant expression plasmids were constructed and transfected into human primary chondrocytes. The expression and subcellular distribution of SLC26A2 protein in chondrocytes were detected by immunoblotting and immunofluorescence. Effects of these variants on chondrocytes viability and apoptosis were measured by Cell Counting Kit-8 (CCK-8) assay. Expression of genes related to cartilage homeostasis was subsequently analyzed by quantitative real-time polymerase chain reaction (qRT-PCR). Results: We identified two compound heterozygous variants c.1020_1022delTGT(p.Val341del) and c.1262 T > C(p.Ile421Thr) in the SLC26A2 gene in the patients. Mutant SLC26A2Val341del and SLC26A2Ile421Thr proteins were distributed in relatively few cells and were observed only within the nucleus. The viability of chondrocytes with the SLC26A2 variant group was similar to the wild-type (WT) group. However, the protein expressions of SLC26A2Val341del and SLC26A2Ile421Thr were decreased compared with SLC26A2WT. Expression levels of matrix metallopeptidase 13 (MMP13), α-1 chain of type X collagen (COL10A1), and Runt-related transcription factor 2 (RUNX2) were significantly decreased in the variant group. However, aggrecan (ACAN) expression was higher in the variant group than the WT group. Conclusions: Overall, our data demonstrate that the variants p.Val341del and p.Ile421Thr in SLC26A2 cause MED-4 and that these two variants promote chondrocyte proliferation while inhibiting chondrocyte differentiation. [ABSTRACT FROM AUTHOR]

Active artificial bone substitutes are crucial in bone repair and reconstruction. Calcium phosphate bone cement (CPC) is known for its biocompatibility, degradability, and ability to fill various shaped bone defects. However, its low osteoinductive capacity limits bone regeneration applications. Effectively integrating osteoinductive magnesium ions with CPC remains a challenge. Herein, we developed magnesium malate-modified CPC (MCPC). Incorporating 5% magnesium malate significantly enhances the compressive strength of CPC to (6.18 ± 0.49) MPa, reduces setting time and improves disintegration resistance. In vitro, MCPC steadily releases magnesium ions, promoting the proliferation of MC3T3-E1 cells without causing significant apoptosis, proving its biocompatibility. Molecularly, magnesium malate prompts macrophages to release prostaglandin E2 (PGE2) and synergistically stimulates dorsal root ganglion (DRG) neurons to synthesize and release calcitonin gene-related peptide (CGRP). The CGRP released by DRG neurons enhances the expression of the key osteogenic transcription factor Runt-related transcription factor-2 (RUNX2) in MC3T3-E1 cells, promoting osteogenesis. In vivo experiments using minipig vertebral bone defect model showed MCPC significantly increases the bone volume fraction, bone density, new bone formation, and proportion of mature bone in the defect area compared to CPC. Additionally, MCPC group exhibited significantly higher levels of osteogenesis and angiogenesis markers compared to CPC group, with no inflammation or necrosis observed in the hearts, livers, or kidneys, indicating its good biocompatibility. In conclusion, MCPC participates in the repair of bone defects in the complex post-fracture microenvironment through interactions among macrophages, DRG neurons, and osteoblasts. This demonstrates its significant potential for clinical application in bone defect repair. [ABSTRACT FROM AUTHOR]

[Display omitted] • Synthesis of superparamagnetic iron oxide nanoparticles (SPIONs) by thermal decomposition method. • Integrin-targeted SPIONs for mechanotransduction stimulus in osteogenic differentiation. • Remotely activated mechanotransduction via Arg-Gly-Asp (RGD) functionalized SPIONs. • Magnetic mechanotransduction in human bone marrow mesenchymal stem cells (hBM-MSCs) for osteogenic differentiation. • Endocytic mechanisms of SPIONs in hBM-MSCs. • Up-regulation of osteogenic markers after magnetic activation of mechanotransduction in hBM-MSCs. Nowadays, diseases associated with an ageing population, such as osteoporosis, require the development of new biomedical approaches to bone regeneration. In this regard, mechanotransduction has emerged as a discipline within the field of bone tissue engineering. Herein, we have tested the efficacy of superparamagnetic iron oxide nanoparticles (SPIONs), obtained by the thermal decomposition method, with an average size of 13 nm, when exposed to the application of an external magnetic field for mechanotransduction in human bone marrow-derived mesenchymal stem cells (hBM-MSCs). The SPIONs were functionalized with an Arg-Gly-Asp (RGD) peptide as ligand to target integrin receptors on cell membrane and used in colloidal state. Then, a comprehensive and comparative bioanalytical characterization of non-targeted versus targeted SPIONs was performed in terms of biocompatibility, cell uptake pathways and mechanotransduction effect, demonstrating the osteogenic differentiation of hBM-MSCs. A key conclusion derived from this research is that when the magnetic stimulus is applied in the first 30 min of the in vitro assay, i.e. , when the nanoparticles come into contact with the cell membrane surface to initiate endocytic pathways, a successful mechanotransduction effect is observed. Thus, under the application of a magnetic field, there was a significant increase in runt-related transcription factor 2 (Runx2) and alkaline phosphatase (ALP) gene expression as well as ALP activity, when cells were exposed to RGD-functionalized SPIONs, demonstrating osteogenic differentiation. These findings open new expectations for the use of remotely activated mechanotransduction using targeted magnetic colloidal nanoformulations for osteogenic differentiation by drug-free cell therapy using minimally invasive techniques in cases of bone loss. [ABSTRACT FROM AUTHOR]

With rates growing quickly with age, osteoarthritis (OA) is the most common cause of chronic disability in aging persons. The discomfort and reduced motion associated with osteoarthritis have a significant impact on quality of life, and there is no known solution. Runt-related transcription factor 1(Runx1) has been shown to play a protective role in the development of osteoarthritis by promoting chondrogenesis. We had created models of ageing mice with osteoarthritis by anterior cruciate ligament transection (ACLT) and analyzed the effects of intra-articular injection of adeno-associated virus/Runx1 (AAV/Runx1) on the models. The results showed that the AAV/Runx1-group maintained better articular cartilage integrity and retained more proteoglycan than the OA group after injection of AAV-Runx1. The markers related to pathological changes in cartilage were downregulated, while the markers related to physiological changes in cartilage were upregulated. This suggests that Runx1 may impede OA progression on the knee joint of ageing mice, potentially playing a protective role in OA and becoming a probable treatment target for osteoarthritis among ageing patients in the future. [ABSTRACT FROM AUTHOR]

Ferroptosis of the retinal pigment epithelial (RPE) cells leads to retinal neuron injury and even visual loss. Our study aims to investigate the role of the SET domain with lysine methyltransferase 7/9 (SET7/9) in regulating high glucose (HG)-induced ferroptosis in RPE cells. The cell model was established by HG treatment. The levels of SET7/9 and Sirtuin 6 (SIRT6) were inhibited and Runt-related transcription factor 1 (RUNX1) was overexpressed through cell transfection, and then their levels in ARPE-19 cells were detected. Cell viability and apoptosis was detected. The levels of reactive oxygen species, malondialdehyde, glutathione, ferrous ion, glutathione peroxidase 4, and acyl-CoA synthetase long-chain family member 4 were detected. SET7/9 and trimethylation of histone H3 at lysine 4 (H3K4me3) levels in the RUNX1 promoter region and RUNX1 level in the SIRT6 promoter region were measured. The relationship between RUNX1 and SIRT6 was verified. SET7/9 and RUNX1 were highly expressed while SIRT6 was poorly expressed in HG-induced ARPE-19 cells. SET7/9 inhibition increased cell viability and inhibited cell apoptosis and ferroptosis. Mechanistically, SET7/9 increased H3K4me3 on the RUNX1 promoter to promote RUNX1, and RUNX1 repressed SIRT6 expression. Overexpression of RUNX1 or silencing SIRT6 partially reversed the inhibitory effect of SET7/9 silencing on HG-induced ferroptosis. In conclusion, SET7/9 promoted ferroptosis of RPE cells through the SIRT6/RUNX1 pathway. [ABSTRACT FROM AUTHOR]

Background: Glycitin, generated from the seeds of the legume soybean, has an essential function in antioxidant, obesity inhibition, and wound healing promotion. Furthermore, recent research has shown that it has anti-inflammatory and cartilage-protective properties. Objectives: This study was designed to investigate the osteogenic effects of glycitin and the transforming growth factor-β (TGF-β) signaling pathway on bone marrow mesenchymal stem cells (BMSCs), as well as their interaction. Materials and Methods: We isolated rabbit BMSCs by whole bone marrow adherent method and cultured, identified, and induced differentiation. The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) method was used to determine the proliferation ability of BMSCs with different concentrations of glycitin at different times; reverse transcription-polymerase chain reaction (RT-PCR) was used to detect the expression levels of osteogenesis-related genes runt-related transcription factor 2 (Runx2), collagen type I (Col-1), and osteocalcin (OC) mRNA; and a kit was used to determine the activities of alkaline phosphatase and triglycerides (TG) in BMSCs. A Western blot was used to detect the expression of TGF-β protein in each group. Results: On the fifth day of BMSC primary culture, the morphology of long spindle-shaped cells might be plentiful, dominated by a daisy-like or whirling arrangement. They proliferated rapidly and fused to 70%–80% on the eighth day of culture; the third generation of BMSCs was taken for phenotyping, and the detection results of flow cytometry analysis showed that the surface antigenic markers of BMSCs, CD44, and CD90 were positive, whereas CD34 and CD45 were negative; after osteogenic induction of BMSCs, visible calcified nodules were evident. There was cartilage matrix formation after osteogenic induction; glycitin promoted BMSC proliferation in a time-dependent and dose-dependent manner, and BMSC proliferation increased significantly at 20 µM glycitin for 3–5 days; glycitin increased the expression of Runx2, Col-1, and OC mRNA in BMSCs and increased the activity of alkaline phosphatase (ALP) while decreasing the activity of TG. Meanwhile, 20 µM glycitin dramatically increased the expression of TGF-β. Conclusion: These findings imply that glycitin increases BMSCs proliferation and osteogenic differentiation. By regulating the TGF-β signaling pathway, glycitin modulates the differentiation of BMSCs into osteoblasts. [ABSTRACT FROM AUTHOR]

Bone mechanotransduction is a critical process during skeletal development in embryogenesis and organogenesis. At the same time, the type and level of mechanical loading regulates bone remodeling throughout the adult life. The aberrant mechanosensing of bone cells has been implicated in the development and progression of bone loss disorders, but also in the bone-specific aspect of other clinical entities, such as the tumorigenesis of solid organs. Novel treatment options have come into sight that exploit the mechanosensitivity of osteoblasts, osteocytes, and chondrocytes to achieve efficient bone regeneration. In this regard, runt-related transcription factor 2 (Runx2) has emerged as a chief skeletal-specific molecule of differentiation, which is prominent to induction by mechanical stimuli. Polycystins represent a family of mechanosensitive proteins that interact with Runx2 in mechano-induced signaling cascades and foster the regulation of alternative effectors of mechanotransuction. In the present narrative review, we employed a PubMed search to extract the literature concerning Runx2, polycystins, and their association from 2000 to March 2024. The keywords stated below were used for the article search. We discuss recent advances regarding the implication of Runx2 and polycystins in bone remodeling and regeneration and elaborate on the targeting strategies that may potentially be applied for the treatment of patients with bone loss diseases. [ABSTRACT FROM AUTHOR]

Background: Laryngeal squamous cell carcinoma (LSCC) is one of the most common malignant tumors of the head and neck. Vasculogenic mimicry (VM) is crucial for tumor growth and metastasis and refers to the formation of fluid channels by invasive tumor cells rather than endothelial cells. However, the regulatory mechanisms underlying VM during the malignant progression of LSCC remain largely unknown. Methods: Gene expression and clinical data for LSCC were obtained from the TCGA and Gene GEO (GSE27020) databases. A risk prediction model associated with VM was established using LASSO and Cox regression analyses. Based on their risk scores, patients with LSCC were categorized into high- and low-risk groups. The disparities in immune infiltration, tumor mutational burden (TMB), and functional enrichment between these two groups were examined. The core genes in LSCC were identified using the machine learning (SVM-RFE) and WGCNA algorithms. Subsequently, the involvement of bone morphogenetic protein 2 (BMP2) in VM and metastasis was investigated both in vitro and in vivo. To elucidate the downstream signaling pathways regulated by BMP2, western blotting was performed. Additionally, ChIP experiments were employed to identify the key transcription factors responsible for modulating the expression of BMP2. Results: We established a new precise prognostic model for LSCC related to VM based on three genes: BMP2, EPO, and AGPS. The ROC curves from both TCGA and GSE27020 validation cohorts demonstrated precision survival prediction capabilities, with the nomogram showing some net clinical benefit. Multiple algorithm analyses indicated BMP2 as a potential core gene. Further experiments suggested that BMP2 promotes VM and metastasis in LSCC. The malignant progression of LSCC is promoted by BMP2 via the activation of the PI3K-AKT signaling pathway, with the high expression of BMP2 in LSCC resulting from its transcriptional activation by runt-related transcription factor 1 (RUNX1). Conclusion: BMP2 predicts poor prognosis in LSCC, promotes LSCC VM and metastasis through the PI3K-AKT signaling pathway, and is transcriptionally regulated by RUNX1. BMP2 may be a novel, precise, diagnostic, and therapeutic biomarker of LSCC. [ABSTRACT FROM AUTHOR]

Antioxidant vanillin (4-hydroxy-3-methoxybenzaldehyde) is used as a flavoring in foods, beverages, and pharmaceuticals. Vanillin possesses various biological effects, such as antioxidant, anti-inflammatory, antibacterial, and anticancer properties. This study aimed to investigate the biological activities of vanillin purified from Adenophora triphylla var. japonica Hara on bone-forming processes. Vanillin treatment induced mineralization as a marker for mature osteoblasts, after stimulating alkaline phosphatase (ALP) staining and activity. The bone-forming processes of vanillin are mainly mediated by the upregulation of the bone morphogenetic protein 2 (BMP2), phospho-Smad1/5/8, and runt-related transcription factor 2 (RUNX2) pathway during the differentiation of osteogenic cells. Moreover, vanillin promoted osteoblast-mediated bone-forming phenotypes by inducing migration and F-actin polymerization. Furthermore, we validated that vanillin-mediated bone-forming processes were attenuated by noggin and DKK1. Finally, we demonstrated that vanillin-mediated antioxidant effects prevent the death of osteoblasts during bone-forming processes. Overall, vanillin has bone-forming properties through the BMP2-mediated biological mechanism, indicating it as a bone-protective compound for bone health and bone diseases such as periodontitis and osteoporosis. [ABSTRACT FROM AUTHOR]

Simple Summary: Cleidocranial dysplasia is a rare disease, manifested by anomalies in the skull, face, teeth, thorax, clavicle, extremities, and short stature. The disease is caused by mutations in the RUNX2 gene, which is involved in the differentiation of cells that give rise and formation of bones. In this study, the genetic material of four patients and their families was analyzed, with the purpose of identifying changes in the sequence of the RUNX2 gene, finding three new changes and one change already reported in the literature. One patient presented cataract and damage to the retina of the eye, and data were not reported in other patients. A bioinformatic analysis of the RUNX2 protein was carried out with the aim of predicting mechanisms, such as the function, stability, and conformation of the protein, and of trying to understand its relationship with the variable presentation of signs and symptoms in this condition. These results can be useful in the genetic counselling of patients. Cleidocranial dysplasia (CCD) is an autosomal dominant skeletal dysplasia characterized by persistent open skull sutures with bulging calvaria, hypoplasia, or aplasia of clavicles permitting abnormal opposition of the shoulders; wide public symphysis; short middle phalanx of the fifth fingers; and vertebral, craniofacial, and dental anomalies. It is a rare disease, with a prevalence of 1–9/1,000,000, high penetrance, and variable expression. The gene responsible for CCD is the Runt-related transcription factor 2 (RUNX2) gene. We characterize the clinical, genetic, and bioinformatic results of four CCD cases: two cases within Mexican families with six affected members, nine asymptomatic individuals, and two sporadic cases with CCD, with one hundred healthy controls. Genomic DNA analyses of the RUNX2 gene were performed for Sanger sequencing. Bioinformatics tools were used to predict the function, stability, and structural changes of the mutated RUNX2 proteins. Three novel heterozygous mutations (c.651_652delTA; c.538_539delinsCA; c.662T>A) and a previously reported mutation (c.674G>A) were detected. In silico analysis showed that all mutations had functional, stability-related, and structural alterations in the RUNX2 protein. Our results show novel mutations that enrich the pool of RUNX2 gene mutations with CCD. Moreover, the proband 1 presented clinical data not previously reported that could represent an expanded phenotype of severe expression. [ABSTRACT FROM AUTHOR]

BACKGROUND: Colorectal cancer (CRC) is a digestive tract malignancy microRNAs (miRNAs) have attracted much attention as biomarkers in tumor studies. OBJECTIVE: This work focused on the predictive potential and mechanism of miR-4310 in CRC. METHODS: The miRNA expression profile sets were obtained from the Gene Expression Omnibus (GEO) database, and the appropriate miRNA was screened by GEO2R. The CRC tissues and control tissues of 88 patients with CRC were collected, and the expression of miR-4310 was detected by quantitative real-time PCR, and the efficacy of miR-4310 in diagnosing CRC was evaluated by the receiver operating characteristic curve (ROC). The effects of miR-4310 on the proliferation, migration, and invasion of CRC cells were explored by Cell Counting Kit-8 (CCK-8) and Transwell experiments. Predicting the potential binding sites of miR-4310 and Runt-related transcription factor 1 (RUNX1) by four predictive websites. The relationship between miR-4310 and RUNX1 was confirmed by a double luciferase reporter gene experiment. RESULTS: The bioinformatics analysis found that miR-4310 was differentially expressed in CRC tissues and this finding was certified by the expression of miR-4310 in CRC tissues of collected patients and cultured CRC cell lines. The expression of miR-4310 had a predictive possibility for CRC patients. MiR-4310/RUNX1 pathway had effects on CRC viability, migration, and invasion. CONCLUSION: MiR-4310 had the potential to be a biomarker for early screening of CRC. MiR-4310 and RUNX1 participated in the regulation of CRC cells. [ABSTRACT FROM AUTHOR]

Objectives: Steroid‐induced osteonecrosis of the femoral head (SONFH) is characterized by impaired osteogenesis in bone marrow mesenchymal stem cells (BMSCs). This study investigates the role of lysine‐specific demethylase 5A (KDM5A) in SONFH to identify potential therapeutic targets. Methods: Human BMSCs were isolated and characterized for cell surface markers and differentiation capacity. A SONFH cell model was established using dexamethasone treatment. BMSCs were transfected with KDM5A overexpression vectors or si‐KDM5A, and the expression of KDM5A, miR‐107, runt‐related transcription factor 2 (RUNX2), osteocalcin (OCN), and osteopontin (OPN) was assessed. Alizarin red staining was used to observe mineralization nodules, while alkaline phosphatase activity and cell viability were measured. The enrichment of KDM5A and histone 3 lysine 4 trimethylation (H3K4me3) on the promoters of RUNX2, OCN, and OPN was analyzed. The binding between miR‐107 and KDM5A 3′UTR was validated, and the combined effect of miR‐107 overexpression and KDM5A overexpression on BMSC osteogenic differentiation was evaluated. Results: KDM5A was upregulated in BMSCs from SONFH. Inhibition of KDM5A promoted osteogenic differentiation of BMSCs, associated with increased RUNX2, OCN, and OPN promoters. KDM5A bound to the promoters of RUNX2, OCN, and OPN, leading to reduced H3K4me3 levels and downregulation of their expression. Overexpression of miR‐107 inhibited KDM5A and enhanced BMSC osteogenic differentiation. Conclusion: KDM5A negatively regulates BMSC osteogenic differentiation by modulating H3K4me3 levels on the promoters of key osteogenic genes. miR‐107 overexpression counteracts the inhibitory effect of KDM5A on osteogenic differentiation. These findings highlight the potential of targeting the KDM5A/miR‐107 axis for SONFH therapy. [ABSTRACT FROM AUTHOR]

Overexpression of the Runt-related transcription factor 2 (RUNX2) has been reported in several cancer types, and the C-X-C motif chemokine receptor 4 (CXCR4) has an important role in tumour progression. However, the interplay between CXCR4 and RUNX2 in melanoma cells remains poorly understood. In the present study, we used melanoma cells and a RUNX2 knockout (RUNX2-KO) in vitro model to assess the influence of RUNX2 on CXCR4 protein levels along with its effects on markers associated with cell invasion and autophagy. Osteotropism was assessed using a 3D microfluidic model. Moreover, we assessed the impact of CXCR4 on the cellular levels of key cellular signalling proteins involved in autophagy. We observed that melanoma cells express both RUNX2 and CXCR4. Restored RUNX2 expression in RUNX2 KO cells increased the expression levels of CXCR4 and proteins associated with the metastatic process. The protein markers of autophagy LC3 and beclin were upregulated in response to increased CXCR4 levels. The CXCR4 inhibitor WZ811 reduced osteotropism and activated the mTOR and p70-S6 cell signalling proteins. Our data indicate that the RUNX2 transcription factor promotes the expression of the CXCR4 chemokine receptor on melanoma cells, which in turn promotes autophagy, cell invasiveness, and osteotropism, through the inhibition of the mTOR signalling pathway. Our data suggest that RUNX2 promotes melanoma progression by upregulating CXCR4, and we identify the latter as a key player in melanoma-related osteotropism. [ABSTRACT FROM AUTHOR]

Objectives: We initially explored the link between the differentially expressed long non-coding RNAs (lncRNAs) and the number of regulatory T (Treg) cells by detecting the lncRNA expression profiles in patients with systemic lupus erythematosus (SLE), then analyzed the correlation between Treg-related lncRNAs and the clinical features of SLE patients, predicting the mechanism by which lncRNAs regulate the differentiation and development of Treg cells, and provided new ideas for the treatment of SLE. Methods: Peripheral blood of 9 active SLE patients were collected and mononuclear cells (PBMCs) were extracted; the lncRNA expression profiles of PBMCs were analyzed by whole transcriptome sequencing. Nine healthy people were used as controls to screen the differentially expressed lncRNAs, to analyze the correlation between lncRNAs and Treg cell number. Pearson test was used to analyze the correlation between lncRNAs and the number of Treg cell, and the correlation between Treg-associated lncRNA and SLEDAI score, ESR, C3, and C4 in SLE patients. The targeted genes of Treg-associated lncRNAs were predicted with miRcode and Targetscan databases and coexpression network. Results: There were 240 differentially expressed lncRNAs in SLE patients compared with healthy controls, including 134 highly expressed lncRNAs (p < 0.05) and 106 lowly expressed lncRNAs (p < 0.05). The expression of ANKRD44-AS1 (r = 0.7417, p = 0.0222), LINC00200 (r = 0.6960, p = 0.0373), AP001363.2 (r = 0.7766, p = 0.0138), and LINC02824 (r = 0.7893, p = 0.0114) were positively correlated with the number of Treg cell, and the expression of AP000640.1 (r = − 0.7225, p = 0.0279), AC124248.1 (r = − 0.7653, p = 0.0163), LINC00482 (r = − 0.8317, p = 0.0054), and MIR503HG (r = − 0.7617, p < 0.05) were negatively correlated with the number of Treg cell. Among these Treg-associated lncRNAs, the expression of LINC00482 (r = − 0.7348, p < 0.05) and MIR503 HG (r = − 0.7617, p < 0.05) were negatively correlated with C3. LINC00200, ANKRD44 - AS1, and AP000640.1 related to Treg cells regulate the expression of signal transducer and activator of transcription 5 (STAT5), phospholipase D1 (PLD1), homeodomain-only protein X (HOPX), and runt-related transcription factor 3 (RUNX3) through competitive binding of miRNA or trans-regulatory mechanism, thereby regulating the differentiation and development of Treg cell. Conclusions: The lncRNA expression profiles were changed in SLE patients, the differentially expressed lncRNAs were associated with abnormal number and function of Treg cells in SLE, and Treg-associated lncRNAs were associated with SLE-disease activity, which may affect the expression of STAT5, PLD1, HOPX, RUNX3 and regulate Treg cell function and participate in the pathogenesis and progression of SLE by competitively binding to miRNAs or trans-regulatory mechanism. Key points • Systemic lupus erythematosus (SLE) is an autoimmune disease involving multiple organs and systems. lncRNAs may affect Treg cells function by regulating genes expression, which may be an important pathogenesis of SLE. • This study, taking SLE as an example, preliminarily analyzed the correlation between lncRNA and Treg cells in SLE patients, analyzed the correlation between Treg-related lncRNA and the clinical characteristics of SLE, and speculated that lncRNA could regulate the differentiation and development of Treg cells through competitive combination with miRNA or trans-regulatory mechanisms. • It is possible to target epigenetic therapy for SLE. [ABSTRACT FROM AUTHOR]

Calcific aortic valve disease (CAVD) is characterized by the fibrosis and mineralization of the aortic valve, which leads to aortic stenosis and heart failure. At the cellular level, this is due to the osteoblastic-like differentiation of valve interstitial cells (VICs), resulting in the calcification of the tissue. Unfortunately, human VICs are not readily available to study CAVD pathogenesis and the implicated mechanisms in vitro; however, adipose-derived stromal/stem cells (ASCs), carrying the patient's specific genomic features, have emerged as a promising cell source to model cardiovascular diseases due to their multipotent nature, availability, and patient-specific characteristics. In this study, we describe a comprehensive transcriptomic analysis of tissue-engineered, scaffold-free, ASC-embedded mineralized tissue sheets using bulk RNA sequencing. Bioinformatic and gene set enrichment analyses revealed the up-regulation of genes associated with the organization of the extracellular matrix (ECM), suggesting that the ECM could play a vital role in the enhanced mineralization observed in these tissue-engineered ASC-embedded sheets. Upon comparison with publicly available gene expression datasets from CAVD patients, striking similarities emerged regarding cardiovascular diseases and ECM functions, suggesting a potential link between ECM gene expression and CAVDs pathogenesis. A matrisome-related sub-analysis revealed the ECM microenvironment promotes the transcriptional activation of the master gene runt-related transcription factor 2 (RUNX2), which is essential in CAVD development. Tissue-engineered ASC-embedded sheets with enhanced mineralization could be a valuable tool for research and a promising avenue for the identification of more effective aortic valve replacement therapies. [ABSTRACT FROM AUTHOR]

Rosavin, a phenylpropanoid in Rhodiola rosea's rhizome, and an adaptogen, is known for enhancing the body's response to environmental stress. It significantly affects cellular metabolism in health and many diseases, particularly influencing bone tissue metabolism. In vitro, rosavin inhibits osteoclastogenesis, disrupts F-actin ring formation, and reduces the expression of osteoclastogenesis-related genes such as cathepsin K, calcitonin receptor (CTR), tumor necrosis factor receptor-associated factor 6 (TRAF6), tartrate-resistant acid phosphatase (TRAP), and matrix metallopeptidase 9 (MMP-9). It also impedes the nuclear factor of activated T-cell cytoplasmic 1 (NFATc1), c-Fos, the nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB), and mitogen-activated protein kinase (MAPK) signaling pathways and blocks phosphorylation processes crucial for bone resorption. Moreover, rosavin promotes osteogenesis and osteoblast differentiation and increases mouse runt-related transcription factor 2 (Runx2) and osteocalcin (OCN) expression. In vivo studies show its effectiveness in enhancing bone mineral density (BMD) in postmenopausal osteoporosis (PMOP) mice, restraining osteoclast maturation, and increasing the active osteoblast percentage in bone tissue. It modulates mRNA expressions by increasing eukaryotic translation elongation factor 2 (EEF2) and decreasing histone deacetylase 1 (HDAC1), thereby activating osteoprotective epigenetic mechanisms, and alters many serum markers, including decreasing cross-linked C-telopeptide of type I collagen (CTX-1), tartrate-resistant acid phosphatase 5b (TRACP5b), receptor activator for nuclear factor κ B ligand (RANKL), macrophage-colony-stimulating factor (M-CSF), and TRAP, while increasing alkaline phosphatase (ALP) and OCN. Additionally, when combined with zinc and probiotics, it reduces pro-osteoporotic matrix metallopeptidase 3 (MMP-3), interleukin 6 (IL-6), and tumor necrosis factor α (TNF-α), and enhances anti-osteoporotic interleukin 10 (IL-10) and tissue inhibitor of metalloproteinase 3 (TIMP3) expressions. This paper aims to systematically review rosavin's impact on bone tissue metabolism, exploring its potential in osteoporosis prevention and treatment, and suggesting future research directions. [ABSTRACT FROM AUTHOR]

Periodontal ligament stem cells (PDLSCs) are important mesenchymal stem cells contributing to regenerating lost periodontal tissues and repairing bone defects. Studies on the molecular mechanism affecting the osteogenic differentiation of PDLSCs are necessary. Scopolamine (SCO) is known as a regulator of neural cell damage. The focus of the current study is on unveiling the role of SCO‐mediated molecular mechanism in the osteogenic differentiation of PDLSCs. Through CCK‐8 assay and LDH detection, we confirmed that SCO enhanced the viability of PDLSCs. Moreover, we determined that SCO induced the PDLSCs osteogenic differentiation, according to data of ALP activity measurement and ARS staining. Mechanistically, we performed western blot and identified that SCO could promote the lactylation of runt‐related transcription factor 2 (RUNX2). We also found through rescue assays that knockdown of RUNX2 could reverse the effect of SCO treatment on the osteogenic differentiation of PDLSCs. Further mechanism investigation revealed that lactylation of RUNX2 at K176 site enhances the protein stability of RUNX2 through deubiquitination. Collectively, our present study unveils that SCO stabilizes RUNX2 to promote the osteogenic differentiation of PDLSCs through the lactylation modification of RUNX2. [ABSTRACT FROM AUTHOR]

Background: Alveolar hypercoagulation and fibrinolytic inhibition are mainly responsible for massive alveolar fibrin deposition, which are closely related with refractory hypoxemia in acute respiratory distress syndrome (ARDS). Our previous study testified runt-related transcription factor (RUNX1) participated in the regulation of this pathophysiology in this syndrome, but the mechanism is unknown. We speculate that screening the downstream genes associated with RUNX1 will presumably help uncover the mechanism of RUNX1. Methods: Genes associated with RUNX1 were screened by CHIP-seq, among which the target gene was verified by Dual Luciferase experiment. Then the efficacy of the target gene on alveolar hypercoagulation and fibrinolytic inhibition in LPS-induced ARDS was explored in vivo as well as in vitro. Finally, whether the regulatory effects of RUNX1 on alveolar hypercoagulation and fibrinolytic in ARDS would be related with the screened target gene was also sufficiently explored. Results: Among these screened genes, AKT3 was verified to be the direct target gene of RUNX1. Results showed that AKT3 was highly expressed either in lung tissues of LPS-induced rat ARDS or in LPS-treated alveolar epithelia cell type II (AECII). Tissue factor (TF) and plasminogen activator inhibitor 1 (PAI-1) were increasingly expressed both in lung tissues of ARDS and in LPS-induced AECII, which were all significantly attenuated by down-regulation of AKT3. Inhibition of AKT3 gene obviously ameliorated the LPS-induced lung injury as well as the collagen I expression in ARDS. RUNX1 overexpression not only promoted the expressions of TF, PAI-1, but also boosted AKT3 expression in vitro. More importantly, the efficacy of RUNX1 on TF, PAI-1 were all effectively reversed by down-regulation of AKT3 gene. Conclusion: AKT3 is an important target gene of RUNX1, through which RUNX1 exerted its regulatory role on alveolar hypercoagulation and fibrinolytic inhibition in LPS-induced ARDS. RUNX1/ATK3 signaling axis is expected to be a new target for the exploration of ARDS genesis and treatment. [ABSTRACT FROM AUTHOR]

BACKGROUND: CXC motif chemokine 5 (CXCL5) is a neutrophil activating peptide derived from epithelial cells, which may be involved in arterial diseases. However, there is yet no report on the effect of CXCL5 in vascular calcification. OBJECTIVE: To explore the role of CXCL5 in the vascular calcification of carotid atherosclerosis (CAS). METHODS: (1) Cytological experiment: Mouse vascular smooth muscle cells (VSMCs) were divided into five groups: osteogenic medium group, Vector group (vector, blank plasmid transfected into VSMCs), CXCL5 group (CXCL5 plasmid transfected into VSMCs), si-NC group (CXCL5 negative control siRNA transfected into VSMCs), si-CXCL5 group (CXCL5 siRNA transfected into VSMCs), Vector+LY2157299 group and CXCL5+LY2157299 group (LY2157299 transferred into the cells 24 hours after cell transfection). Alizarin red staining, alkaline phosphatase staining, and calcium content determination were performed to evaluate the osteogenic differentiation level of VSMCs. (2) Animal experiment: Forty-eight ApoE-/- mice were randomly divided into four groups (n=12 per group): Con+siNC group, Con+si-CXCL5 group, CAS+si-NC group and CAS+si-CXCL5 group. Animal models were not prepared in the first two groups, in which si-NC or si-CXCL5 lentivirus was injected into the tail vein; carotid atherosclerosis models were made in the latter two groups, in which si-NC or si-CXCL5 lentivirus was injected into the tail vein. Von Kossa staining and immunohistochemical staining were used to evaluate carotid vascular calcification and the expression of CXCL5 and transforming growth factor-β receptor 1 (TGFBR1) in mice. RESULTS AND CONCLUSION: In the CXCL5 group, the protein level of runt-related transcription factor 2 (RUNX2) was up-regulated and the level of α-smooth muscle actin was down-regulated, in contrary to the findings in the si-CXCL5 group. In addition, CXCL5 overexpression upregulated the level of TGFBR1, while CXCL5 knockdown inhibited the level of TGFBR1. Compared with the Vector group, the intensity of alizarin red staining, alkaline phosphatase activity and calcium content in the CXCL5 group increased significantly (P < 0.05). Compared with the si-NC group, the intensity of alizarin red staining, alkaline phosphatase activity and calcium content in the si-CXCL5 group decreased significantly (P < 0.05). When LY2157299 inhibited TGFBR1 expression, the osteogenic differentiation of VSMCs induced by CXCL5 was reduced. Compared with the Con+si-NC group, the expression of CXCL5 protein in the carotid artery and calcification area in the CAS+si-NC group increased significantly (P < 0.05). Compared with the CAS+si-NC group, the expression of CXCL5 protein in the carotid artery and vascular calcification area in the CAS+si-CXCL5 group decreased significantly (P < 0.05). In addition, compared with the Con+si-NC group, the expression of RUNX2 protein in the carotid artery in the CAS+si-NC group increased significantly (P < 0.05), while the expression of α-smooth muscle actin protein decreased significantly (P < 0.05). Compared with the CAS+si-NC group, the expression of RUNX2 protein in the carotid artery in CAS+si-CXCL5 group decreased significantly (P < 0.05), while the expression of α-smooth muscle actin protein increased significantly (P < 0.05). In conclusion, CXCL5 can induce osteogenic transformation of VSMCs by activating the TGFBR1 pathway, and inhibition of CXCL5 expression is effective in improving carotid arterial calcification in CAS mice. [ABSTRACT FROM AUTHOR]

Fracture healing in the aged is associated with a reduced healing capacity, which often results in delayed healing or non-union formation. Many factors may contribute to this deterioration of bone regeneration, including a reduced 'angiogenic trauma response'. The phosphodiesterase-3 (PDE-3) inhibitor cilostazol has been shown to exert pro-angiogenic and pro-osteogenic effects in preclinical studies. Therefore, we herein analyzed in a stable closed femoral fracture model whether this compound also promotes fracture healing in aged mice. Forty-two aged CD-1 mice (age: 16–18 months) were daily treated with 30 mg/kg body weight cilostazol (n = 21) or vehicle (control, n = 21) by oral gavage. At 2 and 5 weeks after fracture, the femora were analyzed by X-ray, biomechanics, micro-computed tomography (µCT), histology, immunohistochemistry, and Western blotting. These analyses revealed a significantly increased bending stiffness at 2 weeks (2.2 ± 0.4 vs. 4.3 ± 0.7 N/mm) and an enhanced bone formation at 5 weeks (4.4 ± 0.7 vs. 9.1 ± 0.7 mm3) in cilostazol-treated mice when compared to controls. This was associated with a higher number of newly formed CD31-positive microvessels (3.3 ± 0.9 vs. 5.5 ± 0.7 microvessels/HPF) as well as an elevated expression of phosphoinositide-3-kinase (PI3K) (3.6 ± 0.8 vs. 17.4 ± 5.5-pixel intensity × 104) and runt-related transcription factor (RUNX)2 (6.4 ± 1.2 vs. 18.2 ± 2.7-pixel intensity × 104) within the callus tissue. These findings indicate that cilostazol accelerates fracture healing in aged mice by stimulating angiogenesis and the expression of PI3K and RUNX2. Hence, cilostazol may represent a promising compound to promote bone regeneration in geriatric patients. [ABSTRACT FROM AUTHOR]

Background: Alamandine is a newly characterized peptide of renin angiotensin system. Our study aims to investigate the osteo-preservative effects of alamandine, explore underlying mechanism and bring a potential preventive strategy for postmenopausal osteoporosis in the future. Methods: An ovariectomy (OVX)-induced rat osteoporosis model was established for in vivo experiments. Micro-computed tomography and three-point bending test were used to evaluate bone strength. Histological femur slices were processed for immunohistochemistry (IHC). Bone turnover markers and nitric oxide (NO) concentrations in serum were determined with enzyme-linked immunosorbent assay (ELISA). The mouse embryo osteoblast precursor (MC3T3-E1) cells were used for in vitro experiments. The cell viability was analysed with a Cell Counting Kit‑8. We performed Alizarin Red S staining and alkaline phosphatase (ALP) activity assay to observe the differentiation status of osteoblasts. Western blotting was adopted to detect the expression of osteogenesis related proteins and AMP-activated protein kinase/endothelial nitric oxide synthase (AMPK/eNOS) in osteoblasts. DAF-FM diacetate was used for semi-quantitation of intracellular NO. Results: In OVX rats, alamandine alleviated osteoporosis and maintained bone strength. The IHC showed alamandine increased osteocalcin and collagen type I α1 (COL1A1) expression. The ELISA revealed alamandine decreased bone turnover markers and restored NO level in serum. In MC3T3-E1 cells, alamandine promoted osteogenic differentiation. Western blotting demonstrated that alamandine upregulated the expression of osteopontin, Runt-related transcription factor 2 and COL1A1. The intracellular NO was also raised by alamandine. Additionally, the activation of AMPK/eNOS axis mediated the effects of alamandine on MC3T3-E1 cells and bone tissue. PD123319 and dorsomorphin could repress the regulating effect of alamandine on bone metabolism. Conclusion: Alamandine attenuates ovariectomy-induced osteoporosis by promoting osteogenic differentiation via AMPK/eNOS axis. [ABSTRACT FROM AUTHOR]

BACKGROUND: Previous studies have shown that human dental pulp stem cells have good osteogenic differentiation potential and are potential seed cells in bone tissue engineering, and the effect of recombinant human growth hormone on the proliferative osteogenic differentiation of human dental pulp stem cells is still unclear. OBJECTIVE: To explore the effect of recombinant human growth hormone on the proliferation and osteogenic differentiation of human dental pulp stem cells. METHODS: Human dental pulp stem cells were isolated and cultured by tissue block culture method. After screening according to the drug concentration gradient, recombinant human growth hormone containing 10, 100, 250, 500, 1 000 μg/L was selected as the experimental group, and 0 μg/L without recombinant human growth hormone was selected as the control group. CCK-8 detection reagents were used on days 1, 3, 5, and 7 after the drug intervention to detect the proliferation of human dental pulp stem cells. Different concentrations (10, 100, 250, 500, and 1 000 μg/L) of recombinant human growth hormone were added to the osteogenesis induction solution to intervene in human dental pulp stem cells. Alkaline phosphatase activity was detected by alkaline phosphatase staining and semi-quantitative analysis on day 7 of mineralization induction. The mRNA expression levels of osteogenic gene type I collagen, osteocalcin and Runt-related transcription factor 2 were detected by fluorescence quantitative RT-qPCR. Alizarin red staining was performed on day 14 of mineralization induction to detect osteogenic mineralized nodules. RESULTS AND CONCLUSION: (1) CCK-8 assay results showed that from the third day of intervention, the 100, 250, 500, 1 000 μg/L recombinant human growth hormone group could promote the proliferation of human dental pulp stem cells compared with the control group (P < 0.01). (2) The alkaline phosphatase activity of human dental pulp stem cells in the 100, 250, and 500 μg/L recombinant human growth hormone group was significantly increased compared with the control group (P < 0.01). The number of alizarin-stained mineralized nodules in human dental pulp stem cells in the 100, 250 μg/L recombinant human growth hormone group was significantly increased compared with the control group (P < 0.01). Compared with the control group, the mRNA expression of type I collagen and osteocalcin increased in the 250 μg/L recombinant human growth hormone group (P < 0.05, P < 0.01). mRNA expression of Runtassociated transcription factor 2 increased in the 100 and 250 μg/L recombinant human growth hormone groups (P < 0.01). (3) According to the above results, recombinant human growth hormone at a concentration of 250 μg/L is a more suitable concentration to promote the proliferation and osteogenic differentiation of human dental pulp stem cells. Key words: mesenchymal stem cell. [ABSTRACT FROM AUTHOR]

BACKGROUND: Piezo1, a mechanosensitive protein, is tightly connected to osteogenic differentiation, and it has been demonstrated that TAZ has a role in regulating osteogenic differentiation. It is unclear whether TAZ participates in the regulation of osteogenic differentiation of human bone marrow mesenchymal stem cells by Piezo1, so it is crucial to investigate its unique mechanism to prevent osteonecrosis of the femoral head. OBJECTIVE: To elucidate what function Piezo1 plays in osteogenic differentiation and TAZ expression in human bone marrow mesenchymal stem cells. METHODS: The siRNA targeting Piezo1 was constructed and transfected into 293T cells. The silencing efficiency was detected by RT-qPCR. The selected Piezo1- Home-2337 was packaged according to the silencing efficiency, and its optimal multiplicity of infection value was assayed by immunofluorescence staining. The packaged Piezo1 silencing recombinant lentivirus was transfected into human bone marrow mesenchymal stem cells, and its silencing effect was detected by RT-qPCR and western blot assay. Alizarin red staining, alkaline phosphatase activity analysis, immunofluorescence staining, RT-qPCR and western blot assay were utilized to analyze the effect of silencing Piezo1 on the osteogenic differentiation of human bone marrow mesenchymal stem cells. RESULTS AND CONCLUSION: (1) The mRNA and protein levels of Piezo1 in human bone marrow mesenchymal stem cells transfected by si-Piezo1 were decreased significantly, with a statistically significant difference compared with normal and negative control groups. (2) The alkaline phosphatase activity in the si-Piezo1 group was much lower and the calcium deposition in the si-Piezo1 group was significantly reduced compared with the negative control group. (3) The mRNA levels of osteogenesis-related genes including Runt-related transcription factor 2 (Runx2), osteopontin (OPN), distal-less homeobox 5 (DLX5), osteocalcin, β-catenin and Tafazzin (TAZ) in the si-Piezo1 group were significantly decreased compared with the negative control group. Afterward, the expression levels of TAZ and β-catenin protein in the si-Piezo1 group were down-regulated significantly compared with the negative control group, whereas the expression levels of p-TAZ and p-β-catenin protein in the si-Piezo1 group had the opposite condition. (4) The results of immunofluorescence staining showed that the expression of TAZ and β-catenin in human bone marrow mesenchymal stem cells in the si-Piezo1 group was less compared with the negative control group. (5) These findings indicate that Piezo1 can promote the osteogenic differentiation of human bone marrow mesenchymal stem cells. The osteogenic ability of human bone marrow mesenchymal stem cells is significantly reduced after silencing Piezo1, and the expression of TAZ is also reduced. [ABSTRACT FROM AUTHOR]

Maxillofacial bone defects are treated by autografting or filling with synthetic materials in various forms and shapes. Electrospun nanobiomaterials are becoming popular due to their easy placement and handling; combining ideal biomaterials extrapolates better outcomes. We used a novel electrospun cotton-like fiber made from two time-tested bioresorbable materials, β-TCP and PLLA/PGA, to check the feasibility of its application to maxillofacial bone defects through an in vivo rat mandibular bone defect model. Novel β-TCP/PLLA/PGA and pure β-TCP blocks were evaluated for new bone regeneration through assessment of bone volume, inner defect diameter reduction, and bone mineral density. Bioactive/osteoconductivity was checked by scoring the levels of Runt-related transcription factor x, Leptin Receptor, Osteocalcin, and Periostin biomarkers. Bone regeneration in both β-TCP/PLLA/PGA and β-TCP was comparable at initial timepoints. Osteogenic cell accumulation was greater in β-TCP/PLLA/PGA than in β-TCP at initial as well as late phases. Periostin expression was more marked in β-TCP/PLLA/PGA. This study demonstrated comparable results between β-TCP/PLLA/PGA and β-TCP in terms of bone regeneration and bioactivity, even with a small material volume of β-TCP/PLLA/PGA and a decreased percentage of β-TCP. Electrospun β-TCP/PLLA/PGA is an ideal nanobiomaterial for inducing bone regeneration through osteoconductivity and bioresorbability in bony defects of the maxillofacial region. [ABSTRACT FROM AUTHOR]

Background: Osteoporosis (OP) is a progressive metabolic disorder that is difficult to cure clinically. The molecular mechanisms of OP urgently need to be further examined. This study was designed to explore the potential function of circ_0027885 during osteogenic differentiation, as well as the systematic interactions among circ_0027885, miR-203-3p and runt-related transcription factor 2 (RUNX2). Methods: Relative levels of circ_0027885, miR-203-3p and RUNX2 were analyzed with RT-qPCR and western blotting. Alizarin red staining was performed to detect the mineralization ability under the control of circ_0027885 and miR-203-3p. Dual-luciferase reporter gene assay was conducted to examine the combination among circ_0027885, miR-203-3p and RUNX2. Results: Our research demonstrated that circ_0027885 was significantly increased during hBMSCs differentiation. Overexpression of circ_0027885 notably facilitated osteogenic differentiation and upregulated RUNX2 expression, while knockdown of circ_0027885 reversed the above results. Through prediction on bioinformatics analysis, miR-203-3p was the target binding circ_0027885, and RUNX2 was the potential target of miR-203-3p. Subsequently, these changes induced by the overexpression of circ_0027885 were reversed upon addition of miR-203-3p mimic. Conclusions: Circ_0027885 could sponge miR-203-3p to regulate RUNX2 expression and alleviate osteoporosis progression. [ABSTRACT FROM AUTHOR]

This document provides an overview of the genetic factors, immune responses, and environmental influences involved in ankylosing spondylitis (AS), a rheumatic disorder. It discusses various genes and proteins that play a role in AS, including RANK, TNF-α, CTLA-4, Toll-like receptors, CARD8, Reelin, Calcium-Sensing Receptor, Anthrax Toxin Receptor 2, G protein-coupled receptors, epigenetic changes, and transcription factors. The document emphasizes the complexity of AS and the potential for targeted therapies and improved patient care through a deeper understanding of its genetic mechanisms. [Extracted from the article]

To counteract the effect of zoledronate and decrease the risk of osteonecrosis of the jaw (BRONJ) development in patients undergoing guided bone regeneration surgery, the use of geranylgeraniol (GGOH) has been proposed. Collagen membranes may act as biomimetical drug carriers. The objective of this study was to determine the capacity of collagen-based membranes doped with GGOH to revert the negative impact of zoledronate on the growth and differentiation of human osteoblasts. MG-63 cells were cultured on collagen membranes. Two groups were established: (1) undoped membranes and (2) membranes doped with geranylgeraniol. Osteoblasts were cultured with or without zoledronate (50 μM). Cell proliferation was evaluated at 48 h using the MTT colorimetric method. Differentiation was tested by staining mineralization nodules with alizarin red and by gene expression analysis of bone morphogenetic proteins 2 and 7, alkaline phosphatase (ALP), bone morphogenetic proteins 2 and 7 (BMP-2 and BMP-7), type I collagen (Col-I), osterix (OSX), osteocalcin (OSC), osteoprotegerin (OPG), receptor for RANK (RANKL), runt-related transcription factor 2 (Runx-2), TGF-β1 and TGF-β receptors (TGF-βR1, TGF-βR2, and TGF-βR3), and vascular endothelial growth factor (VEGF) with real-time PCR. One-way ANOVA or Kruskal–Wallis and post hoc Bonferroni tests were applied (p < 0.05). Scanning electron microscopy (SEM) observations were also performed. Treatment of osteoblasts with 50 μM zoledronate produced a significant decrease in cell proliferation, mineralization capacity, and gene expression of several differentiation markers if compared to the control (p < 0.001). When osteoblasts were treated with zoledronate and cultured on GGOH-doped membranes, these variables were, in general, similar to the control group (p > 0.05). GGOH applied on collagen membranes is able to reverse the negative impact of zoledronate on the proliferation, differentiation, and gene expression of different osteoblasts' markers. [ABSTRACT FROM AUTHOR]

(1) To examine the potential mechanism of the Asarum–Angelica drug pair against periodontitis and provide an experimental basis for the treatment of periodontitis with herbal medicine. (2) The core components and core targets of the Asarum–Angelica drug pair in the treatment of periodontitis were detected according to network pharmacology methods. Finally, the effect of the Asarum–Angelica drug pair on osteogenic differentiation was observed in mouse embryonic osteoblast precursor cells. (3) According to the results of network pharmacology, there are 10 potential active ingredients in the Asarum–Angelica drug pair, and 44 potential targets were obtained by mapping the targets with periodontitis treatment. Ten potential active ingredients, such as kaempferol and β-sitosterol, may play a role in treating periodontitis. Cell experiments showed that the Asarum–Angelica drug pair can effectively promote the expression of osteoblast markers alkaline phosphatase (ALP), Runt-related Transcription Factor 2 (RUNX2), and BCL2 mRNA and protein in an inflammatory environment (p < 0.05). (4) Network pharmacology effectively analyzed the molecular mechanism of Asarum–Angelica in the treatment of periodontitis, and the Asarum–Angelica drug pair can promote the differentiation of osteoblasts. [ABSTRACT FROM AUTHOR]

Ovarian cancer is the leading cause of death from gynecologic illnesses worldwide. High-grade serous ovarian cancer (HGSOC) is a gynecological tumor that accounts for roughly 70% of ovarian cancer deaths in women. Runt-related transcription factor 1(RUNX1) proteins were identified with overexpression in the HGSOC. However, the roles of RUNX1 in the development of HGSOC are poorly understood. In this study, combined with whole-transcriptome analysis and multiple research methods, RUNX1 was identified as vital in developing HGSOC. RUNX1 knockdown inhibits the physiological function of ovarian cancer cells and regulates apoptosis through the FOXO1-Bcl2 axis. Down-regulated RUNX1 impairs EMT function through the EGFR-AKT-STAT3 axis signaling. In addition, RUNX1 knockdown can significantly increase the sensitivity to clinical drug therapy for ovarian cancer. It is strongly suggested that RUNX1 work as a potential diagnostic and therapeutic target for HGSOC patients with better prognoses and treatment options. It is possible to generate novel potential targeted therapy strategies and translational applications for serous ovarian carcinoma patients with better clinical outcomes. [ABSTRACT FROM AUTHOR]

Plant extracts are widely used as traditional medicines. Sophora flavescens Aiton-derived natural compounds exert various beneficial effects, such as anti-inflammatory, anticancer, antioxidant, and antiregenerative activities, through their bioactive compounds, including flavonoids and alkaloids. In the present study, we investigated the biological effects of an S. flavescens-derived flavonoid, trifolirhizin (trifol), on the stimulation of osteogenic processes during osteoblast differentiation. Trifol (>98% purity) was successfully isolated from the root of S. flavescens and characterized. Trifol did not exhibit cellular toxicity in osteogenic cells, but promoted alkaline phosphatase (ALP) staining and activity, with enhanced expression of the osteoblast differentiation markers, including Alp, ColI, and Bsp. Trifol induced nuclear runt-related transcription factor 2 (RUNX2) expression during the differentiation of osteogenic cells, and concomitantly stimulated the major osteogenic signaling proteins, including GSK3β, β-catenin, and Smad1/5/8. Among the mitogen-activated protein kinases (MAPKs), Trifol activated JNK, but not ERK1/2 and p38. Trifol also increased the osteoblast-mediated bone-forming phenotypes, including transmigration, F-actin polymerization, and mineral apposition, during osteoblast differentiation. Overall, trifol exhibits bioactive activities related to osteogenic processes via differentiation, migration, and mineralization. Collectively, these results suggest that trifol may serve as an effective phytomedicine for bone diseases such as osteoporosis. [ABSTRACT FROM AUTHOR]

Bone tissue engineering using different scaffolds is a new therapeutic approach in regenerative medicine. This study explored the osteogenic potential of human dental pulp stem cells (hDPSCs) grown on a hydrolytically modified poly(L-lactide-co-caprolactone) (PLCL) electrospun scaffold and a non-woven hyaluronic acid (HYAFF-11™) mesh. The adhesion, immunophenotype, and osteogenic differentiation of hDPSCs seeded on PLCL and HYAFF-11™ scaffolds were analyzed. The results showed that PLCL and HYAFF-11™ scaffolds significantly supported hDPSCs adhesion; however, hDPSCs' adhesion rate was significantly higher on PLCL than on HYAFF-11™. SEM analysis confirmed good adhesion of hDPSCs on both scaffolds before and after osteogenesis. Alizarin red S staining showed mineral deposits on both scaffolds after hDPSCs osteogenesis. The mRNA levels of runt-related transcription factor 2 (Runx2), collagen type I (Coll-I), osterix (Osx), osteocalcin (Ocn), osteopontin (Opn), bone sialoprotein (Bsp), and dentin sialophosphoprotein (Dspp) gene expression and their proteins were higher in hDPSCs after osteogenic differentiation on both scaffolds compared to undifferentiated hDPSCs on PLCL and HYAFF-11™. These results showed that PLCL scaffolds provide a better environment that supports hDPSCs attachment and osteogenic differentiation than HYAFF-11™. The high mRNA of early osteogenic gene expression and mineral deposits observed after hDPSCs osteogenesis on a PLCL mat indicated its better impact on hDPSCs' osteogenic potential than that of HYAFF-11™, and hDPSC/PLCL constructs might be considered in the future as an innovative approach to bone defect repair. [ABSTRACT FROM AUTHOR]

Cleidocranial dysplasia (CCD) is a rare genetic defect caused by a heterozygous mutation of runt-related transcription factor 2 (RUNX2), which is important for osteoblast and skeletal development. RUNX2-deficiency causes extra- and intra-oral malformations that often require orthodontic treatment. Nicotinamide (NAM) affects bone remodelling processes. As these are crucial for orthodontic therapy, NAM could improve orthodontic treatment in CCD patients. This study investigates the effect of NAM in control and RUNX2-deficient osteoblasts under mechanical strain mimicking orthodontic treatment. First, the optimal NAM concentration and the differences in the expression profile of control and RUNX2-deficient osteoblasts were determined. Subsequently, osteoblasts were exposed to tensile and compressive strain with and without NAM, and the expression of genes critically involved in bone remodelling was investigated. NAM increased the expression of bone remodelling genes. RUNX2-deficient osteoblasts expressed more receptor activator of NFkB ligand (RANKL) and interleukin-6 (IL6), but less colony-stimulating factor-1 (CSF1). Most of the positive effects of NAM on bone remodelling genes were impaired by mechanical loading. In conclusion, NAM stimulated osteoblast differentiation by increasing the expression of RUNX2 and regulated the expression of osteoclastogenic factors. However, the positive effects of NAM on bone metabolism were impaired by mechanical loading and RUNX2 deficiency. [ABSTRACT FROM AUTHOR]

Background and Objective: Ankylosing spondylitis is a refractory immune disease that seriously affects the life and work of patients. Epigenetic modifications, especially DNA methylation, have become a research hotspot in complex diseases. We aim to explore the changes in runt‐related transcription factor 2 (RUNX2) gene promoter methylation and transcription level in AS. Method: We detected the RUNX2 gene promoter methylation in 83 AS patients and 83 healthy controls (HCs), then inspected the mRNA difference of RUNX2 between 30 AS patients and 30 HCs by the quantitative real‐time reverse transcription polymerase chain reaction (qRT‐PCR). Results: The RUNX2 gene promoter was hypomethylated in AS patients compared to HCs (p <.001). The research involved 4 CpG regions and 74 CpG sites of RUNX2, of which CpG‐2, CpG‐4 regions, and 18 CpG sites have been differentially methylated. The CpG‐4 island methylation was negatively correlated with C‐reactive protein (p <.05) in AS patients. In the qRT‐PCR validation phase, the mRNA level of RUNX2 in AS patients was significantly higher than HCs (p <.05), and in AS patients who were treated with biologics, the methylation level of CpG‐2 island showed a negative correlation to mRNA (p <.05). ROC results indicated that RUNX2 methylation and its transcription level have good potential to distinguish AS patients from HCs. Conclusion: The RUNX2 gene promoter was hypomethylated in AS patients. Meanwhile, the qRT‐PCR verified the up‐regulated expression on the transcription level, suggesting the abnormal methylation of RUNX2 contributes to the pathogenesis of AS. [ABSTRACT FROM AUTHOR]

The bone marrow (BM) hematopoietic system (HS) gives rise to blood cells originating from hematopoietic stem cells (HSCs), including megakaryocytes (MKs) and red blood cells (erythrocytes; RBCs). Many steps of the cell-fate decision remain to be elucidated, being important for cancer treatment. To explore the role of Wnt/β-catenin for MK and RBC differentiation, we activated β-catenin signaling in platelet-derived growth factor b (Pdgfb)-expressing cells of the HS using a Cre-lox approach (Ctnnb1BM-GOF). FACS analysis revealed that Pdgfb is mainly expressed by megakaryocytic progenitors (MKPs), MKs and platelets. Recombination resulted in a lethal phenotype in mutants (Ctnnb1BM-GOFwt/fl, Ctnnb1BM-GOFfl/fl) 3 weeks after tamoxifen injection, showing an increase in MKs in the BM and spleen, but no pronounced anemia despite reduced erythrocyte counts. BM transplantation (BMT) of Ctnnb1BM-GOF BM into lethally irradiated wildtype recipients (BMT-Ctnnb1BM-GOF) confirmed the megakaryocytic, but not the lethal phenotype. CFU-MK assays in vitro with BM cells of Ctnnb1BM-GOF mice supported MK skewing at the expense of erythroid colonies. Molecularly, the runt-related transcription factor 1 (RUNX1) mRNA, known to suppress erythropoiesis, was upregulated in Ctnnb1BM-GOF BM cells. In conclusion, β-catenin activation plays a key role in cell-fate decision favoring MK development at the expense of erythroid production. [ABSTRACT FROM AUTHOR]

Aim: Fat mass and obesity‐associated (FTO) protein, the first discovered N6‐methyladenine (m6A) demethylase, played positive roles in bone formation. In this study, the aim was to investigate the function and potential mechanism of Fto in dentine formation. Methodology: In vivo model, postnatal 12‐day (PN12), 4‐week‐old (4 wk), 6‐week‐old (6 wk) healthy male C57BL/6J were randomly divided into Fto knockout (Fto−/−) mice and wild‐type (WT) littermates according to their genotypes, with 3–5 mice in each group. The mandibles of Fto−/− mice and WT control littermates were isolated for analysis by micro–computed tomography (micro‐CT), 3‐dimensional reconstruction and Haematoxylin–eosin (HE) staining. In vitro, mouse dental papilla cells (mDPCs) and human dental stem pulp cells (hDPSCs) were cultured with odontogenetic medium to evaluate differentiation capacity; expression levels of odontoblastic related genes were evaluated using quantitative real‐time polymerase chain reaction (qRT‐PCR). The inclusion levels of Runt‐related transcription factor 2 (RUNX2) exon 5 in mDPCs and hDPSCs were detected by semiquantitative real‐time polymerase chain reaction (RT‐PCR). The RNA binding motif protein 4 (RBM4) m6A site was verified through m6A methylated RNA immunoprecipitation (MeRIP) and the stability of RBM4 mRNA influenced by FTO knockdown was measured by mRNA stability assay. Differences with p values <.05 were regarded as statistically significant. Results: We discovered that Fto−/− mice showed significant dentine formation defects characterized by widened pulp cavity, enlarged pulp‐tooth volume ratio, thinned dentine and pre‐dentine layer of root (p <.05). Fto−/− mDPCs and FTO‐silencing hDPSCs not only exhibited insufficient mineralization ability and decreased expression levels of odontoblastic mineralization related genes (p <.05), but showed significantly reduced Runx2 exon 5 inclusion level (p <.05). FTO knockdown increased the m6A level of RBM4 and destabilized the mRNA of RBM4, thus contributing to the reduced RBM4 expression level. Moreover, Rbm4 overexpression in Fto−/− mDPCs can partly restore Runx2 exon 5 inclusion level and the differentiation ability disrupted by Fto knockout. Conclusion: Thus, within the limitations of this study, the data suggest that FTO promotes odontoblastic differentiation during dentine formation by stabilizing RBM4 mRNA to promote RUNX2 exon 5 inclusion. [ABSTRACT FROM AUTHOR]

The present study used network pharmacology and molecular docking to predict the active ingredients and mechanisms of action of Astragalus radix (AR) to promote osteogenic differentiation of bone marrow mesenchymal stem cells (BM‐MSCs), and cell experiments were conducted for verification. First, network pharmacology was used to predict the effective components, targets, and mechanisms of action of AR to promote osteogenic differentiation. The effective components and corresponding target proteins of AR, and the target proteins of osteogenic differentiation were collected through the database. The intersection targets of the two were used for the construction and analysis of a protein–protein interaction (PPI) network. Gene Oncology (GO) and Kyoto Encyclopedia of Genes, and Genomes (KEGG) enrichment analyses were conducted. Next, molecular docking technology was carried out to verify the interaction between the active ingredient and the target protein, and to select the appropriate effective active ingredient. Finally, the results of network pharmacology analysis were verified by in vitro experiments. A total of 95 potential targets were retrieved by searching the intersection of AR and osteogenic differentiation targets. PPI network analysis indicated that RAC‐α‐serine‐threonine‐protein kinase (Akt1) was considered to be the most reliable target for AR to regulate osteogenic differentiation. GO enrichment analysis included 21 biological processes, 21 cellular components and 100 molecular functions. KEGG enrichment analysis indicated that the class I phosphatidylinositol‐3 kinase (PI3K)‐serine‐threonine kinase (Akt) signaling pathway may play an important role in promoting osteogenic differentiation. The results of molecular docking showed that quercetin's performance was improved compared with that of kaempferol. In vitro experiments showed that quercetin promoted the expression of osteogenic marker proteins (including collagen I, Runt‐related transcription factor 2 and osteopontin) in BMSCs and activated the PI3K/Akt signaling pathway. AR acted on Akt1 targets through its main active component quercetin, and promoted the osteogenic differentiation of BM‐MSCs by activating the PI3K/Akt signaling pathway. [ABSTRACT FROM AUTHOR]

Recent evidence suggested that long non-coding RNAs (lncRNAs) and microRNAs (miRNAs) play critical roles in the pathogenesis of vascular calcification (VC). In this study, we tried to explore the expression and role of a lncRNA, i.e., metastasis-associated lung adenocarcinoma transcript 1 (MALAT1), and a miRNA, i.e., miR-30c, in VC. In vitro VC model was induced in human vascular smooth muscle cells (VSMCs) after 10 days culture in calcifying medium containing 2 mM Na2HPO4. Alizarin red S staining, calcium assay and western blot analysis of runt-related transcription factor 2 (Runx2) and alpha smooth muscle actin (α-SMA) were performed to evaluate VC. Knockdown of MALAT1 and up-regulation of MALAT1, miR-30c and Runx2 was performed to determine the impact of these molecules on VSMCs calcification. Dual-luciferase report assay was performed to confirm the relationship between MALAT1 and miR-30c or miR-30c and Runx2. In addition, quantitative reverse transcription PCR and western blot were used to determine gene and protein expression. MALAT1 was increased, while miR-30c was decreased in calcified VSMCs. Knockdown of MALAT1 suppressed VSMCs calcification; on the contrary, up-regulation of MALAT1 promoted VSMCs calcification. The effect of MALAT1 over-expression on VSMCs calcification was reversed by upregulation of miR-30c, which was reversed again by upregulation of Runx2. Dual-luciferase report assay confirmed that there is a direct interaction between MALAT1 and miR-30c, and Runx2 is a direct target of miR-30c. MALAT1 over-expression promoted VSMCs calcification, which was at least partially through regulating the miR-30c/Runx2 axis. [ABSTRACT FROM AUTHOR]

BACKGROUND: The cellular activity of Lin28A can be influenced in several ways, including the self-renewal of embryonic stem cells, somatic cell reprogramming, individual growth and development, tissue metabolism, and carcinogenic consequences. However, neither domestically nor internationally have its effects on periodontal ligament stem cell ability to differentiate into osteoblasts been well explored; the underlying mechanisms are not known. OBJECTIVE: To investigate the impact of Lin28A on periodontal ligament stem cell capacity for osteogenic differentiation. METHODS: By using conventional enzyme digestion, periodontal ligament stem cells were extracted and cultured. The distribution of Lin28A in these cells and the differential expression of Lin28A at different time points of osteogenic induction were detected by qRT-PCR. Lentiviral vectors with Lin28A overexpression and interference expression were constructed and transfected into periodontal ligament stem cells. Fluorescence microscopy was used to observe the transfection effect. qRT-PCR was used to verify the expression difference of Lin28A after lentivirus transfection. The expression levels of osteoblastic genes alkaline phosphatase, Runt-related transcription factor 2 and osteopontin, alkaline phosphatase staining and alizarin red staining results were further detected from positive and negative aspects. The protein expression level of Runt-related transcription factor 2 was detected by western blot assay. Bioinformatics techniques and dual luciferase were used to predict and verify the relationship between Lin28A-related let-7 family members. The periodontal ligament stem cells differentially expressing Lin28A were cultured through osteoblastic induction, and the miRNA with the most obvious expression change during osteoblastic differentiation was determined. RESULTS AND CONCLUSION: (1) Lin28A was enriched in the nucleus of periodontal ligament stem cells, and the expression of Lin28A increased at different time points after osteogenic induction. (2) In the periodontal ligament stem cell models with Lin28A overexpression, alkaline phosphatase staining and alizarin red staining were significantly deeper than those in the blank control group; the expression levels of alkaline phosphatase, Runt-related transcription factor 2 and osteopontin were increased 3.3, 5.1 and 2.2 times, respectively, compared with the blank ones (P < 0.01). In cell models that interfered with Lin28A expression, alkaline phosphatase staining and alizarin red staining were also decreased compared with the control group. The levels of alkaline phosphatase, Runt-related transcription factor 2 and osteopontin decreased 0.19-fold, 0.61-fold and 0.1-fold, respectively (P < 0.01). (3) The protein level of Runt-related transcription factor 2 was consistent with the mRNA difference tendency. The expression of Runt-related transcription factor 2 was increased in the overexpression group and decreased in the interference group. (4) Bioinformatics predicted that Lin28A was associated with eight let-7 family members, which were let-7a, let-7b, let-7c, let-7d, let-7e, let-7f, let-7g and let-7i, respectively. After screening, the expression levels of let-7a and let-7c showed significant changes with Lin28A expression, and let-7a showed the most significant trend. Double luciferase binding assay confirmed that let-7a was directly bound to the 3'-UTR of Lin28A and was involved in the osteogenic differentiation of periodontal ligament stem cells. (5) The results suggested that Lin28A was involved in the regulation of osteogenic differentiation of periodontal ligament stem cells, with a positive correlation. That was, overexpression of Lin28A promoted the osteogenic ability of periodontal ligament stem cells, and the osteogenic ability of periodontal ligament stem cells was weakened after interference of Lin28A expression. The osteogenic differentiation of periodontal ligament stem cells was regulated by Lin28A . [ABSTRACT FROM AUTHOR]

Evidence suggests that enhancing the osteogenic ability of bone marrow-derived mesenchymal stem cells (BMSCs) may be beneficial in the fight against osteoporosis (OP) effects. Inokosterone (IS) is a major active constituent of Achyranthis bidentatae radix (ABR), which stimulates osteogenic differentiation of mouse embryonic osteoblasts. This study aims to investigate effect of IS on OP using osteogenic differentiated BMSCs and ovariectomy (OVX)-induced OP rats. The BMSCs were treated with 50, 100, or 200 mg/L IS and OP rats were given 2 or 4 mg/kg of IS by gavage. Cell viability, the osteogenic differentiation marker protein expression level, and mineralization were observed. This study proved that IS improved cell viability, osteogenic differentiation, and cellular mineralization in BMSCs and raised expression levels of bone morphogenetic protein-2 (BMP2), Smad1, runt-related transcription factor 2 (RUNX2), collagen I, ALP, and OCN. By BMP2 knockdown/overexpression, this study also proved the BMP2 signaling pathway activation is a potential biological mechanism of IS to improve osteogenic differentiation and mineralization in osteogenic differentiated BMSCs. In OVX-induced OP rats, IS was observed to antagonize bone loss, improve osteogenic differentiation marker protein expression levels, and activate BMP-2, smad1, and RUNX2. These findings provide scientific support for further investigation of the biological mechanisms of IS in ameliorating OP. • In vitro , Inokosterone (IS) improved viability and osteogenic differentiation of BMSCs. • In vivo , IS antagonized bone loss of ovariectomy (OVX)-induced OP rats. • BMP2 knockdown/overexpression suggested that IS improve osteoporosis through BMP-2. [ABSTRACT FROM AUTHOR]

Poor osseointegration and postoperative bacterial infections are the main causes of clinical failure of titanium (Ti) based implants. In this study, calcium (Ca) and strontium (Sr), which have osteogenic activity, were simultaneously introduced into titanium dioxide nanotubes (TN) in order to balance the potential cytotoxicity of GL13K (Antibacterial peptides), and a multifunctional Sr, Ca and GL13K-doped TN (SrCaTN-GL13K) coating was successfully constructed. The results showed that the SrCaTN-GL13K coating exhibited good mechanical properties, hydrophilicity, corrosion resistance and bioactivity. Sr2+, Ca2+ and GL13K could be continuously released from the coating and was pH-responsive. In addition, SrCaTN-GL13K showed good antibacterial properties against E. coli and S. aureus with antibacterial rates of approximately 70.32% and 70.78%, respectively. Cellular assays showed that RAW264.7 (immune), human umbilical vein endothelial cell (HUVEC, angiogenic) and mouse embryo osteoblast precursor cell (MC3T3-E1, osteogenic) exhibited good adhesion, survival and proliferation activities on the SrCaTN-GL13K coated surface. At both gene level and protein level, expression of anti-inflammatory markers (Interleukin-10, IL-10; mannose receptor, CD206), angiogenic markers (vascular endothelial growth factor, VEGF; platelet endothelial cell adhesion molecule-1, CD31) and osteogenic markers (alkaline phosphatase, ALP; runt-related transcription factor 2, RUNX2; human collagen type I, COL1a1; osteocalcin, OCN) were significantly upregulated in SrCaTN-GL13K coating. At the cellular and molecular level, SrCaTN-GL13K possessed the ability to induce RAW264.7 towards M2 polarization and it possessed the ability to induce HUVEC tube-formation; meanwhile, it exhibited the ability to induce MC3T3-E1 towards osteogenic differentiation. In conclusion, Ti implants exhibited improved antibacterial, anti-inflammatory, angiogenic, and osteoinductive capabilities by loading Sr, Ca, and GL13K to alter TN. In this study, a multifunctional modification of Ti surface was achieved by an economical and efficient method, which provides a new idea for the design of antimicrobial implantable devices with immunomodulatory functions in the clinic. [ABSTRACT FROM AUTHOR]

Previous studies have suggested a role of phosphatidylinositol‐3‐kinase gamma (PI3Kγ) in bone remodeling, but the mechanism remains undefined. Here, we explored the contribution of PI3Kγ in the resorption of maxillary bone and dental roots using models of orthodontic tooth movement (OTM), orthodontic‐induced inflammatory root resorption, and rapid maxillary expansion (RME). PI3Kγ‐deficient mice (PI3Kγ−/−), mice with loss of PI3Kγ kinase activity (PI3KγKD/KD) and C57BL/6 mice treated with a PI3Kγ inhibitor (AS605240) and respective controls were used. The maxillary bones of PI3Kγ−/−, PI3KγKD/KD, and C57BL/6 mice treated with AS605240 showed an improvement of bone quality compared to their controls, resulting in reduction of the OTM and RME in all experimental groups. PI3Kγ−/− mice exhibited increased root volume and decreased odontoclasts counts. Consistently, the pharmacological blockade or genetic deletion of PI3K resulted in increased numbers of osteoblasts and reduction in osteoclasts during OTM. There was an augmented expression of Runt‐related transcription factor 2 (Runx2) and alkaline phosphatase (Alp), a reduction of interleukin‐6 (Il‐6), as well as a lack of responsiveness of receptor activator of nuclear factor kappa‐Β (Rank) in PI3Kγ−/− and PI3KγKD/KD mice compared to control mice. The maxillary bones of PI3Kγ−/− animals showed reduced p‐Akt expression. In vitro, bone marrow cells treated with AS605240 and cells from PI3Kγ−/− mice exhibited significant augment of osteoblast mineralization and less osteoclast differentiation. The PI3Kγ/Akt axis is pivotal for bone remodeling by providing negative and positive signals for the differentiation of osteoclasts and osteoblasts, respectively. [ABSTRACT FROM AUTHOR]

Simple Summary: The search for regenerative treatments is one of the main objectives of researchers to find effective ways of repairing bone defects and forming areas of bone of sufficient quality and quantity which can receive osseointegrated implants to reconstruct areas affected by periodontal lesions and/or systemic diseases. Biphasic bioceramics are synthetic bone substitutes made of hydroxyapatite (HA) and β-tricalcium phosphate (β-TCP). These materials have osteoconductive properties and are chemically and structurally similar to bone matrix, providing better safety and predictability during guided bone regeneration. This study aimed to evaluate the bone repair process using a new biphasic bioceramic (Plenum® Osshp—70HA: 30β-TCP) compared to a synthetic graft (Straumann® BoneCeramic™) in a critical defect in the calvaria of rats. It was observed that the new Plenum® Osshp bioceramic, in the rat calvaria model, offered excellent safety and surgical predictability, being efficient in the bone repair process. Plenum® Osshp behaved similarly to the commercial control (BoneCeramic™) and can be considered an effective option as a synthetic bone substitute in the dental clinic, especially for implantology. (1) Background: Biphasic bioceramics are synthetic bone substitutes that provide greater safety and better predictability in guided bone regeneration. This study aimed to evaluate the bone repair process using a new biphasic bioceramic of synthetic origin (Plenum® Osshp—70HA: 30β-TCP) in critical calvarial defects. (2) Methods: seventy-four defects were created in rat calvaria and divided into two groups—Plenum® Osshp (PO), right side, and Straumann® BoneCeramic™ (BC), left side. Euthanasia was performed at 7, 15, 30, and 60 days after surgery. (3) Results: Lower gene expression was observed for runt-related transcription factor 2 (RUNX2) and vascular endothelial growth factor (VEGF) and higher expression for Integrin Binding Sialoprotein (IBSP). The results correlated with moderate immunolabeling for osteocalcin (OCN) and slight immunolabeling for osteopontin (OPN) in the PO group. Histometry showed a greater amount of biomaterial remaining in the PO group at 60 days. The microtomographic analysis showed a lower density of bone connectivity and a greater thickness of the trabeculae for the remnants of the PO group. (4) Conclusions: the Plenum® Osshp showed no differences compared to BoneCeramic™ and is therefore considered an effective option as a synthetic bone substitute in bone regeneration. [ABSTRACT FROM AUTHOR]

MHC associations of ankylosing spondylitis in East Asians are complex and involve non-HLA-B27 HLA contributions. Ankylosing spondylitis (AS), a subtype of seronegative spondyloarthropathy, is a common inflammatory disease that affects the spine and sacroiliac joints of the pelvis, resulting in pain, stiffness, and eventually, joint fusion. (B) Percentage of identified major histocompatibility complex (MHC) and non-MHC loci linked with AS. The complicated genetics of AS were decoded using genome-wide association studies (GWAS).[[6]] Through GWAS, approximately 116 major histocompatibility complex (MHC) loci and 48 non-MHC loci have been linked with AS (Figure 1B). [Extracted from the article]

Osteoarthritis (OA) is the most common form of joint disease affecting articular cartilage and peri-articular tissues. Traditional treatments are insufficient, as they are aimed at mitigating symptoms. Multipotent Stromal Cell (MSC) therapy has been proposed as a treatment capable of both preventing cartilage destruction and treating symptoms. While many studies have investigated MSCs for treating OA, therapeutic success is often inconsistent due to low MSC viability and retention in the joint. To address this, biomaterial-assisted delivery is of interest, particularly hydrogel microspheres, which can be easily injected into the joint. Microspheres composed of hyaluronic acid (HA) were created as MSC delivery vehicles. Microrheology measurements indicated that the microspheres had structural integrity alongside sufficient permeability. Additionally, encapsulated MSC viability was found to be above 70% over one week in culture. Gene expression analysis of MSC-identifying markers showed no change in CD29 levels, increased expression of CD44, and decreased expression of CD90 after one week of encapsulation. Analysis of chondrogenic markers showed increased expressions of aggrecan (ACAN) and SRY-box transcription factor 9 (SOX9), and decreased expression of osteogenic markers, runt-related transcription factor 2 (RUNX2), and alkaline phosphatase (ALPL). In vivo analysis revealed that HA microspheres remained in the joint for up to 6 weeks. Rats that had undergone destabilization of the medial meniscus and had overt OA were treated with empty HA microspheres, MSC-laden microspheres, MSCs alone, or a control vehicle. Pain measurements taken before and after the treatment illustrated temporarily decreased pain in groups treated with encapsulated cells. Finally, the histopathological scoring of each group illustrated significantly less OA damage in those treated with encapsulated cells compared to controls. Overall, these studies demonstrate the potential of using HA-based hydrogel microspheres to enhance the therapeutic efficacy of MSCs in treating OA. [ABSTRACT FROM AUTHOR]

BACKGROUND: Less is reported about the effect of transforming growth factor-beta 3 and bone morphogenetic protein-2 on the proliferation and osteogenic differentiation of dental pulp stem cells. OBJECTIVE: To explore the effect of transforming growth factor-beta 3 and bone morphogenetic protein-2 on the osteogenic differentiation of dental pulp stem cells and its preliminary mechanism. METHODS: Dental pulp stem cells were isolated from the pulp tissue of New Zealand rabbit by the enzymatic digestion method. Dental pulp stem cells at passage 3 were divided into the blank group, bone morphogenetic protein-2 group (80 μg/L) and transforming growth factor-beta 3 group (80 μg/L). The cell proliferation activity in each group was tested by MTT assay. After 7 and 14 days of culture, alkaline phosphatase activities were measured. Runt-related transcription factor 2 and bone sialoprotein protein expression levels were conducted by immunocytochemical staining. Osteogenesis-related gene expression was tested through RT-qPCR method. Alizarin red S staining was conducted after 14 and 21 days of culture to observe mineralized nodules. RESULTS AND CONCLUSION: (1) Dental pulp stem cell proliferation activity and alkaline phosphatase activity were higher in the bone morphogenetic protein-2 and transforming growth factor-beta 3 groups than those in the blank group (P < 0.05). The alkaline phosphatase activity of the transforming growth factor-beta 3 group on day 7 was higher than that of the bone morphogenetic protein-2 group (P < 0.05). (2) The expression levels of Runt-related transcription factor 2 and bone sialoprotein in the transforming growth factor-beta 3 group and the bone morphogenetic protein-2 group were higher than those in the blank group (P < 0.05). The protein expression of Runt-related transcription factor 2 in the transforming growth factor-beta 3 group on day 7 was higher than that in the bone morphogenetic protein-2 group (P < 0.05). (3) The expression levels of alkaline phosphatase, type I collagen A1, Runt-related transcription factor 2, osteocalcin, osteopontin and Osx genes in the transforming growth factor-beta 3 group and bone morphogenetic protein-2 group were higher than those in the blank group (P < 0.05). On day 7, the expression levels of alkaline phosphatase, type I collagen A1, and Runt-related transcription factor 2 genes of transforming growth factor-beta 3 group were higher than those of bone morphogenetic protein-2 group (P < 0.05). (4) The mineralized nodules in the transforming growth factor-beta 3 group and bone morphogenetic protein-2 group were more obvious than those in the blank group. (5) The results show that transforming growth factor-beta 3 and bone morphogenetic protein-2 can improve the proliferation and osteogenic differentiation of dental pulp stem cells. The osteogenic induction ability of transforming growth factor-beta 3 on dental pulp stem cells was better than that of bone morphogenetic protein-2 at the early stage of osteogenesis. [ABSTRACT FROM AUTHOR]

BACKGROUND: In recent years, the use of circular RNA (circRNA) to regulate stem cell differentiation has become a new research focus. Therefore, loading circRNA on bone marrow mesenchymal stem cells to study their osteogenic properties is of great significance for the development of new materials to repair bone defect. OBJECTIVE: To investigate the osteogenic effect of circRNA mmu_circ_0001775 knockdown on bone marrow mesenchymal stem cells. METHODS: Bioinformatics analysis tools speculated that the upstream circRNA most closely related to miR-335-5p was mmu_circ_0001775. Lentivirus and negative control virus knockdown mmu_circ_0001775 were transfected into bone marrow mesenchymal stem cells. qRT-PCR was used to detect the expression levels of Runt-related transcription factor 2, osteocalcin, miR-335-5p, and mmu_circ_0001775 genes at 7 and 14 days of osteogenic induction, respectively, to verify their osteogenic effects. Western blot assay was used to detect the expression levels of bone morphogenetic protein 2 and osteopontin. RESULTS AND CONCLUSION: The expression levels of Runt-related transcription factor 2 and osteocalcin at each time point in LV-mmu_circ_0001775-BMMSCs group were higher than those in LV-BMMSCs group (P < 0.01). The relative expression levels of bone morphogenetic protein 2 and osteopontin proteins were significantly higher in the LV-mmu_circ_0001775-BMMSCs group than those of LV-BMMSCs group (P < 0.05). The results showed that cirCRNA mmu_ circ_0001775 knockdown could enhance the osteogenic ability of bone marrow mesenchymal stem cells. [ABSTRACT FROM AUTHOR]

Oncogenic K-RAS mutations occur in approximately 25% of human lung cancers and are most frequently found in codon 12 (G12C, G12V, and G12D). Mutated K-RAS inhibitors have shown beneficial results in many patients; however, the inhibitors specifically target K-RASG12C and acquired resistance is a common occurrence. Therefore, new treatments targeting all kinds of oncogenic K-RAS mutations with a durable response are needed. RUNX3 acts as a pioneer factor of the restriction (R)-point, which is critical for the life and death of cells. RUNX3 is inactivated in most K-RAS-activated mouse and human lung cancers. Deletion of mouse lung Runx3 induces adenomas (ADs) and facilitates the development of K-Ras-activated adenocarcinomas (ADCs). In this study, conditional restoration of Runx3 in an established K-Ras-activated mouse lung cancer model regressed both ADs and ADCs and suppressed cancer recurrence, markedly increasing mouse survival. Runx3 restoration suppressed K-Ras-activated lung cancer mainly through Arf-p53 pathway-mediated apoptosis and partly through p53-independent inhibition of proliferation. This study provides in vivo evidence supporting RUNX3 as a therapeutic tool for the treatment of K-RAS-activated lung cancers with a durable response. [ABSTRACT FROM AUTHOR]

BACKGROUND: Chinese medicine with its good treatment effect and safety is increasingly applied to the treatment of osteoporosis. Bushen Zhuanggu Fang is currently widely used in clinical practice as an agreement of the Affiliated Hospital of Shandong University of Chinese Medicine, and has obtained excellent treatment results. OBJECTIVE: To evaluate the effects of Bushen Zhuanggu Fang on bone metabolism and bone density in a rat model of ovariectomized osteoporosis. METHODS: Sixty Sprague-Dawley rats were randomly grouped into sham group, model group, alendronate sodium group, high-, middle-, and low-dose Bushen Zhuanggu Fang groups (n=10 per group). Rat models of ovariectomized osteoporosis were made in the latter five groups. The sham group received no intervention except for the removal of periovarian adipose tissue. After 8 weeks of modeling, high-, middle- and low-dose Bushen Zhuanggu Fang groups were given 2.34, 1.17, 0.58 g/kg per day, respectively, the alendronate sodium group was given 1 mg/kg per day, and sham operation group and model group were intragastrically given the same volume of normal saline, once a day. The serum levels of alkaline phosphatase, RUNT-related transcription factor 2 and receptor activator of nuclear factor-κB ligand were detected after 12 weeks of continuous gavage. The bone mineral density and microstructure of the right femur were observed. Hematoxylin-eosin staining was used to observe histopathological changes. The expressions of Wnt1 and bone morphogenetic protein 2 were immunohistochemically detected. RESULTS AND CONCLUSION: Compared with the sham group, the bone mineral density and bone trabecular number of the model group were significantly reduced at 8 weeks after modeling (P < 0.05), indicating the successful modeling. Compared with the model group, the bone mineral density, bone volume ratio, bone surface area/tissue volume ratio, and the number of bone trabeculae were significantly increased, while trabecular separation was significantly reduced in the low-, middle-, and high-dose Bushen Zhuanggu Fang groups and the alendronate sodium group (P < 0.01). Moreover, the alendronate sodium group showed better improvements than the three Bushen Zhuanggu Fang groups. Hematoxylin-eosin staining results showed that compared with the model group, the number and continuity of bone trabeculae were increased in the low-, middle- and high-dose Bushen Zhuanggu Fang groups and alendronate sodium group, indicating that Bushen Zhuanggu Fang could significantly improve the bone microstructure of ovariectomized rats. The serum levels of RUNT-related transcription factor 2 in the low-, middle- and high-dose Bushen Zhuanggu Fang groups and the alendronate sodium group were higher than those in the model group (P < 0.01), while the levels of alkaline phosphatase and receptor activator of nuclear factor-κB ligand were significantly lower than those in the model group (P < 0.01). The alendronate sodium group showed better improvement effects than the three Bushen Zhuanggu Fang groups. Immunohistochemistry results showed that the relative expressions of Wnt1 and bone morphogenetic protein 2 in the femur of rats were significantly increased in the low-, middle- and high-dose Bushen Zhuanggu Fang groups and the alendronate sodium group. To conclude, Bushen Zhuanggu Fang can partially improve bone metabolism imbalance in rats, improve the levels of bone formation-related factors, and thereby improve the imbalance between bone resorption and bone formation. [ABSTRACT FROM AUTHOR]

Studies in recent years have highlighted an elaborate crosstalk between T cells and bone cells, suggesting that T cells may be alternative therapeutic targets for the maintenance of bone homeostasis. Here, it is reported that systemic administration of low‐dose staphylococcal enterotoxin C2 (SEC2) 2M‐118, a form of mutant superantigen, dramatically alleviates ovariectomy (OVX)‐induced bone loss via modulating T cells. Specially, SEC2 2M‐118 treatment increases trabecular bone mass significantly via promoting bone formation in OVX mice. These beneficial effects are largely diminished in T‐cell‐deficient nude mice and can be rescued by T‐cell reconstruction. Neutralizing assays determine interferon gamma (IFN‐γ) as the key factor that mediates the beneficial effects of SEC2 2M‐118 on bone. Mechanistic studies demonstrate that IFN‐γ stimulates Janus kinase/signal transducer and activator of transcription (JAK–STAT) signaling, leading to enhanced production of nitric oxide, which further activates p38 mitogen‐activated protein kinase (MAPK) and Runt‐related transcription factor 2 (Runx2) signaling and promotes osteogenic differentiation. IFN‐γ also directly inhibits osteoclast differentiation, but this effect is counteracted by proabsorptive factors tumor necrosis factor alpha (TNF‐α) and interleukin 1 beta (IL‐1β) secreted from IFN‐γ‐stimulated macrophages. Taken together, this work provides clues for developing innovative approaches which target T cells for the prevention and treatment of osteoporosis. [ABSTRACT FROM AUTHOR]