Osteoblast Differentiation Research Articles

Low concentration of quercetin promotes BDNF expression and osteoblast differentiation during fracture healing via TrkB-ERK1/2 signaling pathway.

Quercetin (Que), one of the flavonoids, plays a role in fracture healing, while brain-derived neurotrophic factor (BDNF) and tyrosine kinase receptor kinase B (TrkB) have also been shown to be involved. Que and BDNF signaling pathways are interrelated in the nervous system, but their reciprocal regulatory mechanisms in fracture healing and osteoblast differentiation have not yet been studied. We conducted cellular experiments and fracture animal models to preliminarily clarify the roles and signaling mechanisms of Que and BDNF in osteoblast differentiation and fracture healing. Cellular experiments confirmed that low concentrations of Que (0.01-5μM) promoted osteoblast differentiation and the expression of osteogenesis-related markers, including COL1A1, ALP and Runx2; activated EKR1/2 and promoted the expression of BDNF (P<0.05); and that the combination of Que and exogenous BDNF had the strongest effect on promoting osteogenic differentiation (P<0.05); further mechanistic studies revealed that, the ERK1/2-specific chemical inhibitor PD98059 significantly inhibited the expression of BDNF and osteogenic differentiation markers under the action of Que (P < 0.05), whereas the TrkB-specific chemical inhibitor K252a inhibited the activation of ERK1/2, the expression of BDNF, and the expression of osteoblast differentiation markers under the effect of Que (P < 0.05). In fracture animal models, Que (100mg/kg) significantly promoted fracture healing and increased BDNF expression at the bone callus. Thus, low concentrations of Que promote osteogenic differentiation through the TrkB-ERK1/2-BDNF signaling pathway, which in turn affects fracture healing.

Effect of clindamycin on human osteoblasts treated with zoledronate: An in vitro study.

Effect of clindamycin on human osteoblasts treated with zoledronate: An in vitro study.

GPx-mimetic selenium-enriched yeast nanozymes ameliorate diabetic bone disease via dual-targeting of ROS scavenging and angiogenesis-osteogenesis coupling

GPx-mimetic selenium-enriched yeast nanozymes ameliorate diabetic bone disease via dual-targeting of ROS scavenging and angiogenesis-osteogenesis coupling

Research progress of connexin 43 mediated gap junction communication regulating bone metabolism in glucocorticoid-induced osteonecrosis of the femoral head.

Research progress of connexin 43 mediated gap junction communication regulating bone metabolism in glucocorticoid-induced osteonecrosis of the femoral head.

Nitric oxide synthases: A delicate dance between bone regeneration and neuronal birth.

Nitric oxide synthases: A delicate dance between bone regeneration and neuronal birth.

Comparative Dental Pulp Stem Cells (DPSCs) and Periodontal Ligament Stem Cells (PDLSCs): Difference in effect of aspirin on osteoblast potential of PDLSCs and DPSCs.

Comparative Dental Pulp Stem Cells (DPSCs) and Periodontal Ligament Stem Cells (PDLSCs): Difference in effect of aspirin on osteoblast potential of PDLSCs and DPSCs.

Long noncoding RNA hottip maintained skeletal homeostasis via suppressing the enhancer of zeste homolog 2 (Ezh2)/histone methylation regulatory axis.

Long noncoding RNA hottip maintained skeletal homeostasis via suppressing the enhancer of zeste homolog 2 (Ezh2)/histone methylation regulatory axis.

Dynamic ion-releasing biomaterials actively shape the microenvironment to enhance healing.

Dynamic ion-releasing biomaterials actively shape the microenvironment to enhance healing.

The impact of mechanical unloading on the gut microbiota and the mitigating role of butyrate in bone loss.

The impact of mechanical unloading on the gut microbiota and the mitigating role of butyrate in bone loss.

Intake of eggshell membrane enhances bone mass and suppresses bone marrow adiposity in normal growing rats.

Intake of eggshell membrane enhances bone mass and suppresses bone marrow adiposity in normal growing rats.

Bioactive deproteinized bovine bone mineral based on self-assembled albumin nanoparticles promoted bone regeneration via activation of Wnt/β-catenin pathway.

Bioactive deproteinized bovine bone mineral based on self-assembled albumin nanoparticles promoted bone regeneration via activation of Wnt/β-catenin pathway.

Enhanced osteogenic differentiation of human periodontal ligament cells by mature osteoclasts.

Enhanced osteogenic differentiation of human periodontal ligament cells by mature osteoclasts.

Insights into the potential role of BMSCs-exo delivered USP14 on SIRT1 deubiquitination in Staphylococcus aureus-induced model of osteomyelitis

Osteomyelitis resulting from a traumatic fracture is a recurrent and difficult-to-treat bone infection. Ubiquitin-specific protease 14 (USP14), a deubiquitinating enzyme, and Sirtuin-1 (SIRT1), an NAD+-dependent deacetylase, both play critical roles in regulating cellular processes, including inflammation. It has been discovered that exosomes originated from bone marrow mesenchymal stem cells (BMSCs-exo) can promote the repair and regeneration of bone fractures. In this study, we aimed to investigate the role of BMSCs-exo in osteoblast differentiation in osteomyelitis and the related molecular mechanisms. MC3T3-E1 cells induced with S. aureus were used as an in vitro model of osteomyelitis. BMSCs-exo were isolated and characterized using ultracentrifugation, transmission electron microscopy (TEM), and Western blot. RT-qPCR, Western blot, CCK-8, ALP staining, ELISA, and CO-IP were utilized to evaluate USP14 and SIRT1 levels, the osteogenic differentiation ability of MC3T3-E1 cells, and the deubiquitination level of SIRT1. Low expression of USP14 and SIRT1 was observed in the bone tissue of osteomyelitis patients. BMSCs-exo could upregulate the expression of USP14 and promote the expression of SIRT1 protein in the cell model of osteomyelitis. In addition, BMSCs-exo reduced the levels of inflammatory factors TNFα and IL-6, enhanced cell viability, promoted the expression of osteogenic differentiation markers RUNX2 and OCN in MC3T3-E1 cells, and improved cell osteogenic capacity. However, these trends were significantly reversed in MC3T3-E1 cells following treatment with BMSCs-exo transfected with si-USP14. Furthermore, knockdown of USP14 promoted SIRT1 ubiquitination and degradation, the process that was reversed by the proteasome inhibitor MG132, whereas USP14 overexpression inhibited SIRT1 ubiquitination. In MC3T3-E1 cells infected with S. aureus, BMSCs-exo delivers USP14, which may enhance SIRT1 deubiquitination and increase SIRT1 protein activity. This process inhibits inflammation and promotes osteogenesis, warranting further investigation into its mechanisms and in vivo efficacy.

Chondrocyte maturation bridges two cross-inhibitory subnetworks of the skeletal cell gene regulatory network.

The mechanisms by which crucial transcription factors of a gene regulatory network (GRN) interact continue to be revealed. In the vertebrate skeleton, SOX9 and RUNX2 combine to specify three different cell types. Sox9 drives immature chondrocyte differentiation, Runx2 regulates osteoblast differentiation, and both Sox9 and Runx2 are somehow required for mature chondrocyte formation. To elucidate mechanisms of GRN regulation in mature chondrocytes, transcriptomic data were examined from all three skeletal cell types isolated by laser capture microdissection of embryonic mouse. Multiple bioinformatic analyses supported the hypothesis that SOX9 and RUNX2 operate two cross-inhibitory subnetworks of the skeletal cell GRN during immature chondrocyte and osteoblast formation, but mature chondrocyte differentiation involves cooperation between these subnetworks. Several mature chondrocyte gene clusters had expression levels that represented an averaging of SOX9 and RUNX2 subnetworks, while one cluster, containing the hallmark mature chondrocyte genes collagen type 10a1 and Indian hedgehog, suggested a synergistic interaction between subnetworks. Generally, this in vivo LCM-RNA-seq approach enabled new understanding of interactions between distinct GRN subnetworks during cell differentiation and can similarly reveal regulatory control of any developmental process.

A-485 alleviates postmenopausal osteoporosis by activating GLUD1 deacetylation through the SENP1-Sirt3 signal pathway

ObjectivePostmenopausal osteoporosis (OP) is a bone disease caused by estrogen deficiency. A-485 is a selective inhibitor of p300/CBP histone acetyltransferase (HAT) with potential regulatory effects on bone remodeling. This study aims to investigate the effects of A-485 on postmenopausal OP and its underlying mechanisms.MethodsFor animal experiments, 61 female Wistar rats were used to establish an OP model through ovariectomy (OVX). The rats were administered with A-485 (100 mg/kg/day) via intraperitoneal injection for six weeks. Bone mineral density (BMD) was measured using dual-energy X-ray absorptiometry (DXA). Histopathological changes were observed using HE and Masson’s trichrome staining. ELISA was used to measure bone resorption markers (CTX-1, DPD) and the bone formation marker (P1NP) in rats. Osteoblast differentiation markers (Runx2, OCN), SENP1, Sirt3 expression levels, and GLUD1 acetylation were assessed via Western blot (WB) and RT-qPCR. In vitro, MC3T3-E1 osteogenic progenitor cells were cultured in osteogenic differentiation medium supplemented with ascorbic acid, β-glycerophosphate, dexamethasone, and fulvestrant. CCK-8 was performed to evaluate cell proliferation. Flow cytometry was selected to measure apoptosis and mitochondrial membrane potential. WB and RT-qPCR were employed to analyze ERα, ERβ, Runx2, Sirt3, and GLUD1 acetylation. Additionally, Alizarin red staining was applied to monitor osteoblast mineralization. ATP levels were detected using a commercial kit, and ROS levels were measured by MitoSOX Red.ResultsIn vivo, ovariectomized rats exhibited lower BMD, impaired bone trabeculae, increased CTX-1 and DPD, and altered expression of Runx2 and OCN, all of which were reversed by A-485 treatment. In vitro, A-485 activated GLUD1 deacetylation, enhanced osteogenic differentiation, and improved mitochondrial function. Regarding the mechanism, A-485 activated the SENP1-Sirt3 signal pathway, with SENP1 knockdown negating the effects of A-485. In vivo, A-485 reduced GLUD1 acetylation and promoted improvement of OP, which were reversed by SENP1 knockdown.ConclusionA-485 ameliorates postmenopausal OP by activating GLUD1 deacetylation via the SENP1-Sirt3 signal pathway, thus improving mitochondrial function, and promoting osteogenic differentiation and mineralization.

Spexin-induced MC3T3-E1 cell-derived exosomes enhance osteoblast differentiation.

The roles of exosomes in osteoblast differentiation has been widely investigated. Low exosome production from donor cells constitutes the greatest challenges in exosome-based therapies. Spexin (SPX) is a neuropeptide that is involved in various biological activities including osteogenic differentiation and bone regeneration. Therefore, the purpose of this study was to investigate the effects of SPX on exosome production in osteogenic medium (OM)-treated MC3T3-E1 cells and SPX induced MC3T3-E1 cell-derived exosomes (OM + SPX-Exos) on osteoblast differentiation. To evaluate exosome yield, MC3T3-E1 cells were treated with SPX. Exosome marker expression and particle number were validated via reverse transcriptase-quantitative polymerase chain reaction (RT-qPCR) and nanoparticle tracking analysis (NTA), respectively. MC3T3-E1 cells were then treated with various concentrations of OM + SPX-Exos and osteogenic medium treated MC3T3-E1 derived exosomes (OM-Exos). Cell proliferation, osteogenic differentiation marker expression, alkaline phosphatase (ALP) activity, and mineralization were evaluated using the CCK-8 assay, RT-qPCR, ALP staining, and alizarin red S staining, respectively. SPX significantly increased exosome production and the expression of the exosome markers; Cd63, Rab27a and Alix in MC3T3E1 cells. Furthermore, OM + SPX-Exos significantly increased in the expression of runt-related transcription factor 2(Runx2), alkaline phosphatase, biomineralized associated (Alpl), collagen type I alpha 1 (Col1a1), secreted phosphoprotein 1 (Spp1) and Integrin-binding sialoprotein (Ibsp) at a concentration of 5µg/ml. ALP staining and alizarin red S staining also revealed that OM + SPX-Exos (5 µg/ml) resulted in more ALP-positive cells and markedly promoted mineralization, respectively. In general, these results indicate that SPX stimulates exosome production. OM + SPX-Exos enhances MC3T3-E1 cells proliferation, osteogenic differentiation and mineralization.

Accelerating Sono-Piezoelectric Charge Transport for Antibacterial Therapy and Bone Regeneration by Metal-Deficient TiO2 Spin-Polarization Effect.

Sonodynamic therapy (SDT) is recognized as an effective method for treating deep tissue bacterial infections, but enhancing charge transfer to achieve efficient SDT remains a formidable challenge. Here, deficient TiO2 (DTO) is fabricated and Barium titanate (BTO) heterojunction with oxygen vacancies (OVs) and abundant Ti3+ species on a Ti substrate. The OVs in DTO not only narrow the band gap of TiO2 but also expedite the transfer of surrounding electrons to Ti3+. Meanwhile, the spin-polarization effect induced by the unpaired spin electrons in Ti atoms can expedite the transfer of ultrasonic piezoelectric charges in BTO, enhancing surface sonocatalytic efficiency. Consequently, the DTO/BTO with US irradiation can eradicate 99.82% of Staphylococcus aureus and inhibit biofilm formation. Additionally, the microcurrent generated by ultrasound-excited DTO/BTO enhanced mitochondrial fusion and promoted osteoblastic differentiation. The successful application of this ultrasound-mediated DTO/BTO in treating bone infection defects offers an effective antibiotic-free therapeutic strategy for deep-seated infections.

Emerging nanomaterials capable of effectively facilitating osteoblast maturation.

Efficient osteoblast maturation is essential for successful bone regeneration, yet achieving this goal remains challenging. This review explores the emerging role of nanomaterials in promoting osteoblast differentiation and bone formation. A literature search was conducted in the Web of Science Core Collection in February 2025, covering publications from 2014 to 2024 and limited to articles and proceedings. Keywords included "nanoparticles" and "osteoblast." Among the most extensively studied nanomaterials were hydroxyapatite, carbon-based, and bioactive glass nanoparticles (NPs). These materials influence osteoblast function through intracellular mechanisms, including enhanced mitochondrial activity, autophagy, and osteoinductive gene expression. Additionally, they modulate the extracellular microenvironment by mimicking the native bone matrix, releasing bioactive ions, and reducing inflammation and oxidative stress. Notably, several NP-based systems have reached clinical application, including Signafuse (a bioactive calcium phosphate composite), nanoLOCK (a nanostructured titanium spinal implant), and Vitoss (a synthetic bone graft of nanocrystalline calcium phosphate). More recently, multimodal NPs that integrate different NP types and combine surface roughness, ion release, and chemical cues offer synergistic effects. These materials provide a dual-function approach, targeting both intracellular processes and the bone microenvironment. Their ability to modulate inflammation, oxidative stress, and cellular signaling underscores their translational potential in regenerative medicine and bone tissue engineering.

Sodium Benzoate Inhibits Osteoblast Differentiation and Accelerates Bone Loss by Regulating the FGF2/p38/RUNX2 Pathway.

Sodium benzoate (NaB) is a commonly used food ingredient that is also found in cosmetics and medicines. Previous studies have demonstrated that long-term NaB intake has detrimental effects on human health, while its effects on bone mass remain unknown. In the present study, intragastric NaB administration was found to decrease bone mass and deteriorate bone microstructure in vivo, while prolonged NaB gavage further accelerated bone loss. The in vitro study revealed that NaB inhibited osteoblast differentiation of bone marrow mesenchymal stem cells and MC3T3-E1 cells. Mechanistically, RNA sequencing analysis elucidated that NaB greatly suppressed fibroblast growth factor 2 (FGF2) expression. Further studies revealed that NaB inhibited p38/RUNX2 signaling transduction, which was downstream of FGF2 for modulating osteoblast differentiation. The rescue studies suggested that NaB inhibited RUNX2 expression and osteoblast differentiation through the p38/MAPK signaling pathway. Collectively, NaB accelerated bone loss by inhibiting osteoblast differentiation through downregulating FGF2/p38/RUNX2 signaling pathway. The present study revealed that the long-term intake of NaB-containing food increased the risk of bone loss and osteoporosis (OP). Therefore, a reasonable oral intake of NaB-containing food is an important but convenient initiative for preventing OP.

Humulus lupulus Promoting Osteoblast Activity and Bone Integrity: Effects and Mechanisms.

Osteoporosis is characterized by an imbalance between bone formation and resorption, leading to decreased bone mass and an increased fracture risk, mainly associated with aging. Current treatments include anti-resorptive and anabolic drugs. However, these often have side effects, leading many patients to seek natural biological alternatives. We have demonstrated previously that hops extract, rich in compounds with estrogenic activity classified as phytoestrogens, exerts osteogenic effects by promoting the osteoblastic differentiation of bone marrow stem cells (BMSCs) while inhibiting osteoclast activity in vitro. In our study, young male Sprague Dawley rats were randomized into two groups: one received hops extract (LPL, 1% w/v in drinking water) for two months, and the other drank water alone (C). The rats were euthanized, and their femurs were dissected and processed for static histomorphometry and bone biomechanics. Additionally, BMSCs were isolated from the humeri to evaluate their osteogenic potential. Our result demonstrated that LPL treatment enhanced the osteogenic potential of humeral BMSCs in ex vivo assays, upregulating osteogenic genes and downregulating pro-resorptive markers. These findings correlated with improved femoral bone microarchitecture and biomechanical parameters. In conclusion, a two-month treatment with LPL enhanced the osteogenic capacity of BMSCs, improving bone microarchitecture and biomechanical properties. These results suggest its potential as a natural alternative for promoting bone health.