Osteogenic Differentiation Of MC3T3-E1 Research Articles

Herbal formula xuling-jiangu improves bone metabolic balance in rats with ovariectomy-induced osteoporosis via the gut-bone axis

IntroductionThe XuLing JianGu recipe (XLJGR) is an empirical traditional Chinese medicine formula used for the treatment of osteoporosis. This study aims to explore the effects of XLJGR on the intestinal microbiota composition and endogenous metabolites in ovariectomized (OVX) rats.MethodsAn OVX rat model was established to evaluate the intervention effects of XLJGR. The measured indicators included bone density, serum bone metabolism markers, and an analysis of the types and abundances of intestinal microbiota, along with changes in endogenous metabolites. Additionally, MC3T3-E1 cells were used to validate the differential metabolites.ResultsXLJGR significantly reduced the abundance of Bacteroides, Butyricicoccus, and other bacterial strains in the gut. KEGG metabolic pathway enrichment analysis showed that XLJGR intervention led to notable changes in pathways such as peptidoglycan biosynthesis, carbapenem biosynthesis, and vancomycin resistance. Moreover, XLJGR significantly upregulated key intestinal microbiota metabolites, including gabapentin(GAB), camphoric acid(CAA), and nonanedioic acid(AZA), thereby promoting the proliferation and osteogenic differentiation of MC3T3-E1 cells.DiscussionThis study highlights the potential biomedical applications of XLJGR in promoting bone health by positively affecting intestinal microbiota and metabolic characteristics. These findings suggest that XLJGR may serve as a viable alternative in the treatment of osteoporosis, warranting further exploration of its therapeutic mechanisms and clinical applications.

A 3D printed magnesium ammonium phosphate/polycaprolactone composite membrane for Guided bone regeneration

Guided bone regeneration (GBR) is a widely utilized technique for alveolar bone augmentation and the GBR membrane plays a crucial role in this procedure. However, developing a membrane that possesses both mechanical property and biodegradability still remains a challenge. In this study, MgNH4PO4·6H2O (MNP) and polycaprolactone (PCL) were utilized to prepare a novel MNP-PCL composite GBR membrane via 3D printing method, and five different radios of MNP were included (5, 10, 15, 20, 25 wt%). All samples were well prepared into 25 mm × 25 mm square membranes with a thickness of about 0.8 mm. The membranes exhibited promising interconnected network structure with great distribution uniformity, mechanical properties, and biodegradability. Additionally, the membranes also possessed the capacities of sustained magnesium ions release (4 weeks at least) and weakly alkaline environment maintainance (pH 8.0–––8.5), which might be beneficial for promoting bone formation, this hypothesis was then confirmed by in vitro and in vivo studies that the adhesion, proliferation, osteogenic differentiation of MC3T3-E1 and new bone formation were signicantly enhanced on 10 % and 15 % MNP-PCL surfaces. In summary, the MNP-PCL membrane may providing a new approach for the development of GBR membranes in the future.

DMP1 and IFITM5 Regulate Osteogenic Differentiation of MC3T3-E1 on PEO-Treated Ti-6Al-4V-Ca2+/Pi surface.

Despite numerous studies on various surface modifications on titanium and its alloys, it remains unclear what kind of titanium-based surface modifications are capable of controlling cell activity. This study aimed to understand the mechanism at the cellular and molecular levels and investigate the in vitro response of osteoblastic MC3T3-E1 cultured on the Ti-6Al-4V surface modified by plasma electrolytic oxidation (PEO) treatment. A Ti-6Al-4V surface was prepared by PEO at 180, 280, and 380 V for 3 or 10 min in an electrolyte containing Ca2+/Pi ions. Our results showed that PEO-treated Ti-6Al-4V-Ca2+/Pi surfaces enhanced the cell attachment and differentiation of MC3T3-E1 compared to the untreated Ti-6Al-4V control but did not affect cytotoxicity as shown by cell proliferation and cell death. Interestingly, on the Ti-6Al-4V-Ca2+/Pi surface treated by PEO at 280 V for 3 or 10 min, MC3T3-E1 showed a higher initial adhesion and mineralization. In addition, the alkaline phosphatase (ALP) activity significantly increased in MC3T3-E1 on the PEO-treated Ti-6Al-4V-Ca2+/Pi (280 V for 3 or 10 min). In RNA-seq analysis, the expression of dentin matrix protein 1 (DMP1), sortilin 1 (Sort1), signal-induced proliferation-associated 1 like 2 (SIPA1L2), and interferon-induced transmembrane protein 5 (IFITM5) was induced during the osteogenic differentiation of MC3T3-E1 on the PEO-treated Ti-6Al-4V-Ca2+/Pi. DMP1 and IFITM5 silencing decreased the expression of bone differentiation-related mRNAs and proteins and ALP activity in MC3T3-E1. These results suggest that the PEO-treated Ti-6Al-4V-Ca2+/Pi surface induces osteoblast differentiation by regulating the expression of DMP1 and IFITM5. Therefore, surface microstructure modification through PEO coatings with Ca2+/Pi ions could be used as a valuable method to improve biocompatibility properties of titanium alloys.

MiR-181a-5p promotes osteogenesis by targeting BMP3

High-throughput microRNA (miRNA) sequencing of osteoporosis was analyzed from the Gene Expression Omnibus (GEO) database to investigate specific microRNAs that control osteogenesis. MiR-181a-5p was differentially expressed among healthy subjects and those with osteoporosis. Inhibitors and mimics were transfected into cells to modulate miR-181a-5p levels to examine the role in MC3T3-E1 functions. Alkaline phosphatase (ALP) staining and Alizarin Red S (ARS) staining were used for morphological detection, and proteins of ALP and Runt-related transcription factor 2 (RUNX2), as osteogenesis markers, were detected. During the osteogenic differentiation of MC3T3-E1, the transcription level of miR-181a-5p was significantly increased. The inhibition of miR-181a-5p suppressed MC3T3-E1 osteogenic differentiation, whereas its overexpression functioned oppositely. Consistently, the miR-181a-5p antagomir aggravated osteoporosis in old mice. Additionally, we predicted potential target genes via TargetScan and miRDB and identified bone morphogenetic protein 3 (BMP3) as the target gene. Moreover, the reduced expression of miR-181a-5p was validated in our hospitalized osteoporotic patients. These findings have substantial implications for the strategies targeting miR-181a-5p to prevent osteoporosis and potential related fractures.

In Situ Growth of Tunable Gold Nanoparticles by Titania Nanotubes Templated Electrodeposition for Improving Osteogenesis through Modulating Macrophages Polarization.

Driving macrophages M2 polarization has attracted growing attention for improving osteogenesis. Here, the in situ growth of tunable gold nanoparticles (AuNPs) on titania nanotubes (TiNTs) array was realized by electrodeposition, with the guidance of TiNTs. The fabricated Au layer showed excellent biocompatibility with different osteoimmune effects. Briefly, the Au deposition on 5 and 10 V anodized TiNTs surface could induce RAW264.7 cells to M2 polarization, whereas the Au deposition on 20 V anodized TiNTs surface showed M1 polarization, as indicated by various markers determination through immunofluorescence staining, qPCR, Western blot, and ELISA. Furthermore, the osteogenic differentiation of MC3T3-E1 was significantly enhanced by the macrophages conditioned medium from the Au@10VNTs surface. The in vivo tests also confirmed denser and thicker new trabecula bone formation and more M2 macrophages infiltration both on and adjacent to the Au@10VNTs implant surface. In mechanism, the cytokine array analysis of macrophages conditioned medium from the Au@10VNTs surface revealed the upregulation of pro-healing cytokines such as IL-10 and VEGF and downregulation of pro-inflammatory cytokines such as IL-1β and MCSF. In addition, the NF-κB pathway was significantly inhibited. In conclusion, the electrodeposition of a Au layer guided by TiNTs is a promising strategy for reducing postoperative inflammatory reactions and improving osseointegration through modulating macrophages polarization.

Silymarin prevents iron overload induced bone loss by inhibiting oxidative stress in an ovariectomized animal model

Silibinin (SIL) has been used extensively for its hepatoprotective properties and antioxidant properties, including bone health. Iron overload can inhibit osteogenic proliferation and differentiation and promote bone loss. However, whether SIL can reverse the harmful effects of iron overload inovariectomized (OVX) rats and the mechanism is not clear. Therefore, this study intends to investigate the effect of SIL on bone mass and bone metabolism in iron overload rats and also explore the role of SIL on osteogenic differentiation of MC3T3-E1.RT-qPCR was used to measure the transcribe of target genes. Furthermore, alizarin red staining, alkaline phosphatase staining, immunofluorescence and CCK-8 assay were conducted to detect cell viability and target protein expression, osteogenic function. The OVX rat model with iron overload was set up to investigate bone reconstruction.Our results demonstrated that SIL promotes the proliferation and differentiation of osteoblasts, increases the ALP secretion and mineralization ability of osteoblasts, and enhances the transcribe and expression of target genes including OC, Runx-2, SOD2 and SIRT1 in an iron overload environment. In addition, it was confirmed that systemic SIL administration inhibits bone loss in OVX rats with iron overload and changes bone metabolism and oxidative stress status. Further study has shown that iron overload exerts its harmful function by accelerating bone turnover-mediated changes in higher bone metabolism to worsen osteoporosis. SIL can inhibit the unfriendly effects of iron overload, and by modifying bone metabolism and oxidative stress levels, the results contribute to clinical prevention and treatment of the progression of postmenopausal osteoporosis.

Modulation of T Cell Responses by Fucoidan to Inhibit Osteogenesis.

Fucoidan has sparked considerable interest in biomedical applications because of its inherent (bio)physicochemical characteristics, particularly immunomodulatory effects on macrophages, neutrophils, and natural killer cells. However, the effect of fucoidan on T cells and the following regulatory interaction on cellular function has not been reported. In this work, the effect of sterile fucoidan on the T-cell response and the subsequent modulation of osteogenesis is investigated. The physicochemical features of fucoidan treated by high-temperature autoclave sterilization are characterized by UV–visible spectroscopy, X-ray diffraction, Fourier transform infrared and nuclear magnetic resonance analysis. It is demonstrated that high-temperature autoclave treatment resulted in fucoidan depolymerization, with no change in its key bioactive groups. Further, sterile fucoidan promotes T cells proliferation and the proportion of differentiated T cells decreases with increasing concentration of fucoidan. In addition, the supernatant of T cells co-cultured with fucoidan greatly suppresses the osteogenic differentiation of MC3T3-E1 by downregulating the formation of alkaline phosphatase and calcium nodule compared with fucoidan. Therefore, our work offers new insight into the fucoidan-mediated T cell and osteoblast interplay.

Involvement of MiRNA-211-5p and Arhgap11a Interaction During Osteogenic Differentiation of MC3T3-E1 Cells.

ObjectiveMicroRNAs (miRNAs) are well-recognized for their abilities to regulate gene expression post-transcriptionally in plants and animals. Recently, miRNA-messenger RNA (mRNA) regulatory relationships have been confirmed during biological processes, including osteogenic differentiation. This study aimed to find out more candidate miRNA-mRNA pairs involved in the osteogenic differentiation of MC3T3-E1 cells.MethodsAn MC3T3-E1-based microarray dataset (accessioned as GSE46400) downloaded from the Gene Expression Omnibus included MC3T3-E1 cells with or without 14-day osteoblast differentiation osteoblast induction. Multiple miRNA-mRNA prediction databases were searched by differentially expressed genes (DEGs) to obtain pairs of a miRNA-DEG regulatory network. The MC3T3-E1 cells were cultured and incubated in the osteogenic differentiation medium for 14 days. The expressions of candidate miRNAs and mRNAs were determined by real-time quantitative PCR(RT-qPCR) in MC3T3-E1 cells. The miRNA-mRNA interactions were verified by dual-luciferase reporter gene assays and experiments using mimics miRNA or their inhibitors.ResultsWe identified 715 upregulated DEGs and 603 downregulated DEGs between MC3T3-E1 cells with and without osteoblast induction by analyzing the raw data of the GSE46400 dataset. There were 7 overlapped miRNA-mRNA pairs identified during osteogenic differentiation of MC3T3-E1 cells, including mmu-miR-204-5p-Arhgap11a, mmu-miR-211-5p-Arhgap11a, mmu-miR-24-3p-H2afx, mmu-miR-3470b-Chek2, mmu-miR-3470b-Dlgap5, mmu-miR-466b-3p-Chek1, and mmu-miR-466c-3p-Chek1. The Arhgap11a, H2afx, Chek2, Dlgap5, and Chek1 were hub genes downregulated in MC3T3-E1 cells after osteogenic differentiation, verified by RT-qPCR results. The RT-qPCR also determined declined expressions of miR-204-5p and miR-24-3p concomitant with elevated expressions of miR-211-5p, miR-3470b, miR-466b-3p, and miR-466c-3p in the MC3T3-E1 cells, with osteoblast induction compared with undifferentiated MC3T3-E1 cells. Dual-luciferase reporter gene assays demonstrated Arhgap11a as the target of miR-211-5p. MiR-211-5p upregulation by its mimic increased Arhgap11a expression in MC3T3-E1 cells.ConclusionOur study characterizes miR-211-5p targeting Arhgap11a promotes the osteogenic differentiation of MC3T3-E1 cells, which provides novel targets to promote the osteogenesis process during bone repair.

Arctiin elevates osteogenic differentiation of MC3T3-E1 cells by modulating cyclin D1

ABSTRACT Osteoporosis is a systemic disorder of bone metabolism. This study aimed to investigate the impacts and possible mechanisms of Arctiin, a lignin isolated from Arctium lappa on MC3T3-E1 osteoblast differentiation. In this study, after treatment with different concentrations of Arctiin, reverse transcription-quantitative polymerase chain reaction (RT-qPCR) and western blotting were used to estimate the expression of osteogenesis markers. Then, the activity of alkaline phosphatase (ALP) was detected by an ALP assay kit and calcium nodules staining was evaluated by alizarin red staining (ARS). Additionally, the regulatory effects of Arctiin on cyclin D1 (Ccnd1) was assessed by measurement of protein expression. Subsequently, the functions of Ccnd1 silencing on the osteogenic differentiation was examined in Arctiin-treated MC3T3-E1 cells. Results indicated that Arctiin dose-dependently upregulated the expression of runt-related transcription factor 2 (RUNX2), collagen type 1 (COL1A1), osteocalcin (OCN) and osteopontin (OPN). Elevated ALP activity and calcification degree was prominently observed in the Arctiin-treated groups. Moreover, Ccnd1 expression was notably enhanced after Arctiin intervention. Importantly, Ccnd1-knockdown abrogated the impacts of Arctiin on osteogenic differentiation of MC3T3-E1. To conclude, findings in this study suggested that Arctiin could regulate MC3T3-E1 osteoblast differentiation via up-regulating Ccnd1, supporting that Arctiin might be a therapeutic target for osteoporosis.

Fluoride induces hypomethylation of BMP2 and activates osteoblasts through the Wnt/β-catenin signaling pathway

BackgroundSkeletal fluorosis has become a public health issue in recent years as its serious impact on patients’ life expectancy. Bone morphogenetic protein 2 (BMP2) plays a key role in promoting osteogenesis. However, the mechanism of BMP2-Wnt/β-catenin axis in skeletal fluorosis needs further exploration. MethodsThe RT-qPCR and western blot assay were carried out to examine the mRNA and protein levels. Cell viability was measured by MTT assay. A commercial ALP assay kit was used to detect ALP activities. Alizarin Red staining was performed to measure the formation of mineralized nodules. Methylation-specific PCR (MSP) was performed to measure the methylation level of BMP2. ResultsFluoride promoted the expression of osteogenic marker genes (OPN, OCN, OSX and RUNX2) and induced the proliferation and differentiation of MC3T3-E1 cells. Fluoride induced hypomethylation and high expression of BMP2. Furthermore, knockdown of BMP2 reversed the promoting effect of fluoride on osteogenic differentiation of MC3T3-E1. The expression of β-catenin, glycogen synthase kinase 3β (GSK3β), wingless/integrated 3α (Wnt3α), low-density lipoprotein receptor-related protein 5 (LRP5) and dishevelled 1 (Dv1) were increased in osteoblasts treated with fluoride, however, knockdown of BMP2 reversed this phenomenon. Simultaneous knockdown of BMP2 and β-catenin significantly inhibited the differentiation of osteoblasts induced by fluoride. ConclusionFluoride contributed to proliferation and differentiation of osteoblasts through BMP2-Wnt/β-catenin axis, providing a feasible theoretical basis for the treatment of skeletal fluorosis.

The effects and mechanism of paeoniflorin in promoting osteogenic differentiation of MC3T3-E1

BackgroundThe incidence of osteoporosis and osteoporotic fractures is increasing every year. Traditional Chinese Medicine (TCM) can shed new light on the treatment of osteoporosis. This study aimed to explore the role and mechanism of paeoniflorin in promoting osteogenic differentiation of an osteoblast precursor cell line (MC3T3-E1).MethodsMC3T3-E1 cells were cultured in osteogenic induction medium (OIM) and OIM combined with different concentrations of paeoniflorin. The optimal dose of paeoniflorin was assessed by a cell counting kit-8 (CCK-8) assay. Then, alkaline phosphatase (ALP) and Alizarin Red S (ARS) staining were performed to assess the osteogenic capacity of paeoniflorin. The transcription of osteogenic genes and the expression of osteogenic proteins were assessed by RT-PCR and Western blotting, respectively. The transcription of Wnt/β-catenin signaling pathway genes and proteins was assessed by RT-PCR and Western blotting, respectively. Finally, Dickkopf-1 (DKK-1), a Wnt/β-catenin signaling pathway inhibitor, was used to identify whether the Wnt/β-catenin signaling pathway was involved in the osteogenic differentiation of paeoniflorin. Osteoclastogenesis in RAW264.7 cells was identified by tartrate-resistant acid phosphatase (TRAP) staining.ResultsAt concentrations ranging from 0.1 to 100 μM, paeoniflorin was not cytotoxic to MC3T3-E1 cells. Paeoniflorin significantly increased the osteogenic differentiation of MC3T3-E1 cells in a dose-dependent manner. Moreover, paeoniflorin significantly increased osteogenic differentiation gene and protein expression. Through bioinformatic analysis, paeoniflorin-affected genes were found to be involved in different signaling pathways, such as the Wnt/β-catenin signaling pathway. Paeoniflorin enhanced β-catenin and CyclinD1 expression compared with that of the control groups. DKK-1 partially reversed the promoting effects of paeoniflorin in promoting osteogenic differentiation of MC3T3-E1 cells. Moreover, paeoniflorin inhibited the osteoclastogenesis of RAW264.7 cells.ConclusionPaeoniflorin promotes osteogenic differentiation in MC3T3-E1 cells by regulating the Wnt/β-catenin pathway. Paeoniflorin is a potential therapeutic agent for the treatment of osteoporosis.

Exosomes from gingival mesenchymal stem cells enhance migration and osteogenic differentiation of pre-osteoblasts.

Gingival mesenchymal stem cells (GMSCs) have great potential in bone tissue regeneration. However, it is not well known how on exosomes derived from GMSCs affect the functions of bone-related cells. In this study, we explored the impact of GMSCs-derived exosomes (GMSCs-Exos) on pre-osteoblast MC3T3-E1 proliferation, migration and osteogenic differentiation. Results of CCK-8 assay showed that GMSCs-Exos had no effect on proliferation of pre-osteoblasts. Further, we found that GMSCs-Exos promoted the migration of pre-osteoblasts and osteogenic differentiation of MC3T3-E1 as revealed by enhanced Alizarin red staining, elevated alkaline phosphatase (ALP) activity and upregulated expression of osteogenic genes. This study provides new insights into the potential exosome-mediated paracrine mechanism of GMSCs in bone regeneration.

2D titanium carbide(MXene) nanosheets and 1D hydroxyapatite nanowires into free standing nanocomposite membrane: in vitro and in vivo evaluations for bone regeneration.

Bone loss or insufficiency remains a great challenge for implant integrated and subsequently functional loading, where developing biomaterials to augment bone quantity and regenerate alveolar bone defects at implant site is vitally necessary. Recently, MXene, as a large new family of 2D materials, exhibits a great prospect in biomedical applications owing to its ultrathin structure and morphology with a range of extraordinary properties such as chemical, electronic, optical and biological properties etc. Besides, hydroxyapatite is a favorable biomaterial with outstanding bioactivity and osteogenic capacity. In this study, we prepared free standing UHAPNWs/MXene nanocomposite membranes via introducing ultralong hydroxyapatite nanowires (UHAPNWs) with different weight ratios into MXene to explore their potential in bone regeneration. SEM, XPS, FTIR, XRD, tensile strength, Young's modulus and water contact angles were used to characterize the morphology, chemical composition, surface properties, mechanical properties and hydrophilicity of the materials. Subsequently, in vitro studies like cell adhesion, proliferation and osteogenic differentiation of MC3T3-E1 were evaluated. The incorporation of UHAPNWs improved mechanical properties and hydrophilicity with an enhancement in cell adhesion, proliferation, and osteogenic differentiation. More importantly, 10wt% UHAPNWs/MXene exhibited the optimal mechanical properties while biological improvement was more pronounced along with the addition of UHAPNWs when the weight fraction of UHAPNWs was from 0 to 30wt%. Furthermore, in vivo experiments the UHAPNWs/MXene nanocomposite membranes effectively enhanced bone tissue formation quantitatively and qualitatively in a rat calvarial bone defect. Therefore, an appropriate amount of UHAPNWs into MXene plays a positive and evident role in enhancing mechanical properties, biocompatibility and osteoinductivity, leading a novel inorganic composite material for regeneration of bone tissue.

Enhancing Effect of Vitexin on Osteogenic Activity of Murine Pre-Osteoblastic Mc3t3-E1 Cells

Vitexin (5,7,4-trihydroxyflavone-8-glucoside), a natural flavone present in a variety of plants, is well known for its rich pharmacological properties. However, its osteogenic activity remains unclear to date. The purpose of this study was to explore the effects of vitexin on osteogenic activity in murine pre-osteoblastic MC3T3-E1 cells using the MTT assay for cell proliferation, alkaline phosphatase (ALP) activity assay for cell differentiation, and Von Kossa staining for cell mineralization. Quantitative real-time PCR was used for the detection of osteocalcin (OCN) mRNA expression in cells. Furthermore, effects of vitexin on the differentiation and matrix mineralization of dexamethasone (DEX)- suppressed cells was also investigated. The results showed vitexin could significantly enhance cell proliferation in a low concentration range of 10-10-10-6 μg mL-1. ALP activity was significantly increased after the cells were treated with vitexin at 10-8 and 10-6 μg mL-1. The expression levels of the osteogenic OCN gene in cells treated with vitexin at 10-6, 10- 8, and 10-10 μg mL-1 were improved by 3.1-fold, 5.8-fold, and 4.2-fold over the control, respectively. Additionally, vitexin (10-8 μg mL-1) significantly alleviated the inhibitory effect of osteoblast differentiation and mineralization induced by DEX. Collectively, our findings suggest vitexin could enhance cell proliferation and osteogenic differentiation of MC3T3-E1 cells, as well as rescue the inhibitory effect of cell differentiation and matrix mineralization induced by DEX. Therefore, vitexin may be useful as a promising therapeutic agent for bone disease and plays an important role in the prevention of glucocorticoid-induced osteoporosis.

In vitro bioactivity of AH plus with the addition of nano-magnesium hydroxide.

BackgroundAH Plus (AH) has been widely used as a root canal sealer in the endodontic field due to its superior physicochemical properties. However, clinical application of AH is limited due to its weak bioactivity.MethodsIn this study, we have developed an AH cement containing nano-magnesium hydroxide (NMH) as an additive to enhance the bioactivity of AH. The NMH can neutralize pH and facilitate bone formation. The objective of this study was to evaluate the effects of NMH and modified AH on osteoblasts behavior in vitro. The CCK-8, alkaline phosphatase (ALP) staining, and real-time polymerase chain reaction (PCR) assays were used to assess the proliferation and differentiation of MC3T3-E1 cells, respectively. The adhesion and spreading of MC3T3-E1 cells were investigated in vitro by scanning electron microscopy (SEM). Meanwhile, the flow and magnesium ion release of the modified AH was also concerned.ResultsIn vitro cell assays further showed that the addition of NMH into AH cement, which was denoted as modified AH (especially AH+3%NMH), could effectively improve the proliferation and osteogenic differentiation of MC3T3-E1 cells.ConclusionsTaken all together, we believe that the modified AH samples (especially AH+3%NMH) have outstanding biocompatibility and osteogenic properties and may have great potential in endodontic field.

The effect of celecoxib in traumatic heterotopic ossification around temporomandibular joint in mice

In this study, the role of inflammation in traumatic heterotopic ossification around temporomandibular joint (THO-TMJ), as well as the preventive and treatment effect of celecoxib in THO-TMJ both in vivo and in vitro were explored. A surgically-induced THO-TMJ mouse model and a co-culture model of ATDC-5 or MC3T3-E1 and RAW-264.7cells were used in this study for in vivo and in vitro research. A series of inflammatory factors, such as CD3, CD68, CD20, IL-10, IL-6 and TNF-α, were activated 48h after trauma in a THO-TMJ model. Local trauma initiated systemic inflammatory responses as well as T cell- and macrophage-mediated local inflammatory responses around TMJ. In addition, expression of COX-2 was significantly elevated. The findings also showed that local injection of celecoxib could effectively alleviate the inflammatory response around TMJ at the early stage of trauma and inhibit the formation of THO-TMJ in vivo. Meanwhile, celecoxib could inhibit chondrogenic differentiation of ATDC-5 and osteogenic differentiation of MC3T3-E1 under inflammatory condition in vitro. Furthermore, celecoxib could inhibit the expression of Bmpr1b in the injured condylar cartilage at the initiation stage of THO-TMJ, which implied that Bmpr1b expressed by the residual condylar cartilage might be related to the pathogenesis of THO-TMJ. Inflammation played a crucial role in the pathogenesis of THO-TMJ, and anti-inflammation might be a possible choice to inhibit THO-TMJ, which provided scientific clues for the mechanisms, pharmacotherapy and molecular intervention of THO-TMJ.

Zirconia Reinforced Trace Element Co-doped Hydroxyapatite Coating

Zirconia is a kind of prospective hard tissue implant materials, which possesses superior mechanical properties and excellent biocompatibilities. To promote the stable osseointegration between Zirconia implants and tissue, zirconia-toughened trace Sr-, Si- and F- co-doped hydroxyapatite (ZrO2-DHA) was coated by plasma spraying on zirconia. The phase composition and structure of coatings were characterized, and the mechanical properties and in vitro biological properties were investigated. The results show that co-doping of trace Sr, Si and F enhances the biological properties of coatings on adhesion and differentiation of osteoblasts through the signal transduction pathway of osteogenic differentiation. All of ZrO2-DHA coatings promote the cell viability and gene expression in osteogenic differentiation of MC3T3-E1. On the 7th day of cell culture, the relative expression levels of Alp and Col-I in the ZrO2-DHA coating containing 70% DHA (7DHA) were about 2.8 times and 2.3 times higher than those in the ZrO2 substrate group, respectively. Mechanical properties of ZrO2-DHA coatings are improved with the increment of zirconia ratio. Hardness of DHA coating and 7DHA coating are 250.8 and 313.8HV respectively and their bond strength are 25.1 and 31.8 MPa, respectively. 7DHA coatings with network structure possess both excellent mechanical and biological performance for the implant coating application.

Transcriptome analysis of osteoblasts in an ovariectomized mouse model in response to physical exercise.

ObjectivesOsteoporosis is a metabolic disease resulting in progressive loss of bone mass as measured by bone mineral density (BMD). Physical exercise has a positive effect on increasing or maintaining BMD in postmenopausal women. The contribution of exercise to the regulation of osteogenesis in osteoblasts remains unclear. We therefore investigated the effect of exercise on osteoblasts in ovariectomized mice.MethodsWe compared the activity of differentially expressed genes of osteoblasts in ovariectomized mice that undertook exercise (OVX+T) with those that did not (OVX), using microarray and bioinformatics.ResultsMany inflammatory pathways were significantly downregulated in the osteoblasts after exercise. Meanwhile, IBSP and SLc13A5 gene expressions were upregulated in the OVX+T group. Furthermore, in in vitro assay, IBSP and SLc13A5 mRNAs were also upregulated during the osteogenic differentiation of MC3T3-E1 and 7F2 cells.ConclusionThese findings suggest that exercise may not only reduce the inflammatory environment in ovariectomized mice, indirectly suppressing the overactivated osteoclasts, but may also directly activate osteogenesis-related genes in osteoblasts. Exercise may thus prevent the bone loss caused by oestrogen deficiency through mediating the imbalance between the bone resorptive activity of osteoclasts and the bone formation activity of osteoblasts.Cite this article: W-B. Hsu, W-H. Hsu, J-S. Hung, W-J. Shen, R. W-W. Hsu. Transcriptome analysis of osteoblasts in an ovariectomized mouse model in response to physical exercise. Bone Joint Res 2018;7:601–608. DOI: 10.1302/2046-3758.711.BJR-2018-0075.R2.

Vaspin regulates the osteogenic differentiation of MC3T3-E1 through the PI3K-Akt/miR-34c loop

Vaspin regulates the osteogenic differentiation of MC3T3-E1 through the PI3K-Akt/miR-34c loop

Effect of CXCR4 inhibitor AMD3100 on alkaline phosphatase activity and mineralization in osteoblastic MC3T3-E1 cells

The aim of the study was to investigate the effect of C-X-C chemokine receptor type 4 (CXCR4) inhibitor AMD3100 on the osteogenic differentiation of pre-osteoblastic cell line MC3T3-E1. In this study we found that blocking SDF-1/CXCR4 signaling with AMD3100 strongly suppressed osteogenic differentiation in MC3T3-E1 cells, as evidenced by an early decrease in the activity of alkaline phosphatase (ALP), and down-regulation of mRNA expression of the osteogenic master regulator Runx2, ALP, osteocalcin, and progressive ankylosis genes. Moreover, we found that the regulatory effect of AMD3100 might be mediated via intracellular STAT3 activation. However, AMD3100 exerted no significant effect on generation of matrix mineralization at the terminal stage of osteogenic induction. In conclusion, our results demonstrated an inhibitory role of AMD3100 in osteogenic differentiation of MC3T3-E1 cells, especially in the early stage, which provides novel insights into the effect of CXCR4 antagonists on modulation of osteogenesis.