OPN mRNA Research Articles

Effect of Insulin-Like Growth Factor-1 on Bone Formation of Bone Marrow Mesenchymal Stem Cells Under High Glucose Environment by Regulating Insulin Like Growth Factor Binding Protein 2(IGFBP-2)/p38 Pathway

Diabetes easily affects the biological properties of bone marrow mesenchymal stem cells (BMSCs). Insulin-like growth factor-1 (IGF-1) promotes bone healing during osteoporotic fractures. However, IGF-1's effect on BMSCs high glucose is unclear. Rat BMSCs were assigned into control group, high glucose group, and IGF-1 group in which BMSCs were transfected with pc-DNA 3.1-IGF-1 plasmid on the basis of high glucose followed by analysis of IGF-1, RUNX2 and OPN mRNA level by real time PCR, cell proliferation by MTT assay, alkaline phosphatase (ALP) activity, IGFBP-2 level by ELISA, and p38 phosphorylation by Western blot. High glucose group showed significantly decreased IGF-1, RUNX2 and OPN mRNA level, reduced ALP activity, IGFBP-2 expression and p38 phosphorylation compared to control group (P < 0 05). Transfection of IGF-1 plasmid under high-glucose environment significantly upregulated IGF-1, RUNX2 and OPN mRNA, increased ALP activity, IGFBP-2 expression and p38 phosphorylation compared to high-glucose group (P < 0 05). IGF-1 expression is reduced in high glucose environment. Up-regulation of IGF-1 can promote BMSCs osteogenic differentiation through the IGFBP-2/p38 pathway.

Animal Experimental Study on Repair of Alveolar Bone Defect with Double-Pipe Biphase Bioceramic Polyetheretherketone Composite

Polyetheretherketone (PEEK) has the advantages of good biocompatibility, thus become a widely used bone remodeling material. Bioceramics are also highly effective bone repair materials. However, the repairing effect of biphasic bioceramics combined with polyetheretherketone composites on alveolar bone defects has not been elucidated. SD rats were separated into control group that was established as alveolar bone injury model; PEEK group that the alveolar bone injury model was repaired with polyether ether ketone; and composite group that alveolar bone injury model was repaired with double-tube biphase bioceramic/polyetheretherketone. After 8 weeks of treatment, the bone mineral density (BMD) changes were assessed by X-ray absorptiometry. HE staining was used to analyze the changes of tooth defect. The change of alkaline phosphatase (ALP) content was analyzed. Real-time PCR was performed to measure osteogenic factors Opn and Runx2 expressions. Serum BMP-2 level was analyzed by ELISA. The alveolar ridge height was compared and the alveolar bone repair rate was calculated. In the PEEK group and the composite group, BMD was significantly increased, bone was repaired, Runx2 and Opn mRNA expression was upregulated and ALP activity was enhanced along with elevated BMP-2 secretion, alveolar ridge height and bone repair rate compared to control group (P < 0 05). The composite group exhibited more significant changes compared with PEEK group (P < 0 05). Double-tube biphasic bioceramic/polyetheretherketone composites can repair alveolar bone defects, promote osteogenic differentiation, induce BMP-2 secretion, enhance bone density, and accelerate alveolar bone repair.

Homozygous Loss-of-Function Mutations in CCDC134 Are Responsible for a Severe Form of Osteogenesis Imperfecta.

Osteogenesis imperfecta (OI) is a primary bone fragility disorder with an estimated prevalence of 1 in 15,000 births. The majority of OI cases are inherited in an autosomal-dominant manner, while 5% to 10% have recessive or X-linked inheritance. Up to now, approximately 5% of OI cases remain without mutation demonstrated, supporting the involvement of other genes in the disease spectrum. By whole-exome sequencing, we identified a homozygous variant (c.2T>C) in CCDC134 gene in three patients from two unrelated families with severe bone fragility that did not respond to bisphosphonate treatment, short stature, and gracile long bones with pseudarthroses but no dentinogenesis imperfecta. CCDC134 encodes a secreted protein widely expressed and implicated in the regulation of some mitogen-activated protein kinases (MAPK) signaling pathway. Western blot and immunofluorescence analyses confirmed the absence of CCDC134 protein in patient cells compared with controls. Furthermore, we demonstrated that CCDC134 mutations are associated with increased Erk1/2 phosphorylation, decreased OPN mRNA and COL1A1 expression and reduced mineralization in patient osteoblasts compared with controls. These data support that CCDC134 is a new gene involved in severe progressive deforming recessive osteogenesis imperfecta (type III). © 2020 American Society for Bone and Mineral Research.

Effect of Hepatic Kinase B1 (LKB1) on Osteogenic Differentiation of Bone Marrow Mesenchymal Stem Cells During Senescence

Hepatic kinase B1 (LKB1) is a tumor suppressor and regulates cell proliferation and apoptosis. However, whether LKB1 affects bone marrow mesenchymal stem cells (BMSCs) osteogenic differentiation of during aging remains unclear. Two BMSCs derived from Zempster24−/− (aging) and Zempster24+/+ (normal) mice were cultured in vitro followed by measurement of LKB1 expression by real-time quantitative PCR and Western blot. LKB1 siRNA was transfected into Zempster24−/−BMSCs and LKB1 expression was measured. 14 days after osteogenic induction, mineralized nodule formation was evaluated by alizarin red staining, expression of Calcin, type I collagen, RUNX2 and OPN mRNA expression was measured, together with alkaline phosphatase (ALP) activity and the PI3K/mTOR pathway activity. Compared with normal BMSCs, LKB1 expression was significantly increased, calcified nodules were decreased, with reduced expression of osteocalcin, type I collagen, RUNX2 and OPN mRNA as well as decreased ALP activity and PI3K/mTOR signaling protein expression (P < 0.05). LKB1 siRNA transfection into senescent BMSCs down-regulated LKB1 expression, increased calcification nodule formation, expression of osteocalcin, type I collagen, RUNX2 and OPN mRNA, as well as increased ALP activity and PI3K/mTOR pathway protein expression (P < 0.05). Aging can promote the increase of LKB1 expression and inhibit the osteogenic differentiation of BMSCs. Down-regulation of LKB1 expression in BMSCs during senescence can promote osteogenic differentiation through regulating PI3K/mTOR pathway.

Regenerative Effect of Resorbable Scaffold Embedded Boron-Nitride/Hydroxyapatite Nanoparticles in Rat Parietal Bone.

BN has important roles in several physiological events, including bone growth and immune system. New infection-free cranioplasty and has an osteogenic activities material that are compatible with tissue are being developed. We aimed in our study to examine whether different combinations of Boron-nitride/Hydroxyapatite are embedded into the scaffold in the treatment of calvarial defects. 200 adult female Sprague-Dawley rats divided into 10 equal groups. Osteotomy was made by trepan drill in 8 mm diameter. The scaffolds were placed in the rats and were left to recovery for 2 months. During the experiment, CT scans were taken from the calvarial areas of the rats in the 2nd, 4th and 8th weeks. Significant healing was observed in defect diameters in 2.5% BN+10% HA, 2.5% BN and 5% BN+10% HA, respectively. After 8 weeks, it was seen that the amounts of OPN, BMP-2, RunX2 and ALP mRNA expression significantly decreased in 2.5% BN+10% HA, 2.5% BN, 5% BN+10% HA and 5% BN groups. It was shown that bone recovery was at the best grade in the groups, which contained 2.5% BN and 2.5% BN+10% HA when compared to the other groups. BN is a very promising agent that will be used in reconstructive surgery for the treatment of calvarial bone defects.

Interleukin-15 Regulates Osteogenic/Adipogenic Differentiation of Bone Marrow Mesenchymal Stem Cells Under Oxidative Stress by Regulating PI3K/Akt/mTOR Signaling Pathway

The oxidative stress process can affect bone marrow mesenchymal stem cells (BMSCs) differentiation. Interleukin (IL-15) regulates fat differentiation of BMSCs. However, the role of IL-15 in osteogenic/adipogenic differentiation of BMSCs under oxidative stress remains unclear. Rat BMSCs were cultured and randomly divided into control group; oxidative stress group and IL-15 treatment group followed by analysis of IL-15 secretion by ELISA, expression of osteocalcin, type I collagen, RUNX2 and OPN mRNA as well as FABP4 and PPARγ 2 by Real time PCR, ALP activity, ROS content and SOD activity, and expression of PI3K/Akt/mTOR signaling proteins by Western blot. In oxidative stress group, IL-15 secretion was significantly decreased, osteocalcin, type I collagen, RUNX2 and OPN mRNA expression was reduced, along with deceased ALP activity and SOD activity, increased ROS content and FABP4 and PPARγ 2 protein expression as well as decreased expression of p-AKT and mTOR in comparison of control (P < 0 05). IL-15 treatment on oxidative stress BMSCs significantly increased IL-15 secretion and the expression of osteocalcin, type I collagen, RUNX2 and OPN mRNA, along with increased ALP activity and SOD activity, decreased FABP4 and PPAR 2 protein expression and ROS content as well as increased expression of p-AKT and mTOR (P < 0 05). IL-15 secretion was reduced in BMSCs under oxidative stress. Promoting IL-15 secretion can improve redox balance through PI3K/Akt/mTOR signaling pathway, promote osteogenic differentiation of BMSCs.

Effects of LncRNA SNHG15 on Osteogenic/Adipogenic Differentiation of Bone Marrow Mesenchymal Stem Cells Under Oxidative Stress by TGFβ/Smad Signaling Pathway

Oxidative stress can affect bone marrow mesenchymal stem cells (BMSCs). LncRNA SNHG15 involves in a variety of cellular physiological and pathological processes. However, LncRNA SNHG15’s effect on osteogenesis/adipogenic differentiation of BMSCs under oxidative stress remains unclear. BMSCs were divided into normal control group; oxidative stress group; SNHG15 group in which SNHG15 plasmid was transfected under oxidative stress condition followed by analysis of the expression of LncRNA SNHG15, RUNX2 and OPN by real-time quantitative PCR, reactive oxygen species (ROS) and superoxide dismutase (SOD) activity, FABP4 and PPARγ2 mRNA expression, alkaline phosphatase (ALP) activity and TGFβ/Smad signaling by Western blot. Under oxidative stress, the expression of SHHG15, RUNX2 and OPN mRNA was significantly decreased, ROS production was increased, SOD activity was reduced, along with increased expression of FABP4 and PPARγ2 protein, decreased ALP activity, as well as reduced expression of TGFβ1, Smad2, and Smad7 compared to control group (P < 0.05). Transfection of LncRNA SNHG15 plasmid significantly up-regulated the expression of SNHG15, RUNX2 and OPN mRNA, decreased ROS production, increased SOD activity, decreased expression of FABP4 and PPARγ2 mRNA, and increased ALP activity as well as increased expression of TGFβ1, Smad2, and Smad7 compared to control group (P < 0.05). The expression of SNHG15 is decreased in BMSCs under oxidative stress. Up-regulation of SNHG15 expression can improve the redox balance through TGFβ/Smad signaling pathway, promote osteogenic differentiation of BMSCs under oxidative stress, and inhibit its differentiation into fat.

Effect of GGCX on the differentiation function of osteoporosis bone marrow mesenchymal stem cells through regulating TGFβ/smad signaling pathway.

Osteoporosis (OP) has a high incidence and can be found in multiple age groups. The bone marrow mesenchymal stem cells (BMSCs) have the potential for self-renewal and multi-directional differentiation, which are often used for investigating the differentiation function of osteoporosis bone marrow mesenchymal stem cells. γ-glutamyl carboxylase (GGCX) is a carboxylase-related carboxylase and was observed to be abnormally expressed in osteoarthritis. However, the role and related mechanisms of GGCX in OP have not been fully elucidated. This work aimed to evaluate the effect of GGCX on the differentiation function of BMSCs. Sprague-Dawley rats were randomly divided into the OP group prepared by ovariectomy and sham group. GGCX expression was tested by enzyme-linked immunosorbent assay (ELISA). BMSCs were isolated from OP rats and transfected with pcDNA-GGCX plasmids. BMSC proliferation was detected by tetrazolium salt colorimetry (MTT) assay. The osteogenic and adipogenic differentiation of BMSCs was analyzed by alizarin red staining and oil red O staining. The ALP activity was determined by alkaline phosphatase (ALP) activity colorimetric assay. Real time-PCR was used to test the expressions of osteogenesis-related genes RUNX2 and OPN mRNA. Western blot was adopted to assess the TGFβ/smad signaling pathway activity. GGCX expression was significantly decreased in the serum of OP rats compared with the sham group (p < 0.05). The transfection of pcDNA-GGCX plasmid significantly promoted BMSC cell proliferation, increased calcified nodule formation, inhibited adipogenic differentiation, enhanced ALP activity, elevated RUNX2, and OPN mRNA expressions, and upregulated TGFβ1, Smad2, and Smad7 expressions (p < 0.05). GGCX secretion is reduced in osteoporosis. GGCX can regulate osteoporosis via promoting the TGFβ/smad signaling pathway, facilitating BMSCs osteogenic differentiation, and inhibiting BMSCs adipogenic differentiation.

Mir-149 Promotes Survival and Differentiation of Bone Marrow Mesenchymal Stem Cells by Regulating Interleukin-6/Signal Transducer and Activator of Transcription 3 Signaling Pathway

Bone marrow mesenchymal stem cells (BMSCs) can self-renew with multi-directional differentiation. Mir-149 is involved in various diseases, but whether Mir-149 regulates the survival and differentiation of BMSCs and related mechanisms remains unclear. BMSCs were isolated and randomly divided into Si-NC group, Mir-149 siRNA group, and Mir-149 siRNA + STAT3 inhibitor WP1066 group followed by analysis of the expression of Mir-149, RUNX2 and OPN mRNA by real time PCR, BMSCs proliferation by MTT assay, Caspase 3 activity, ALP activity, formation of type II collagen and IL-6 level by ELISA, as well as STAT3 signaling pathway expression by Western blot. Mir-149 expression was reduced in BMSCs of Mir-149 siRNA group, with promoted survival of BMSCs, decreased Caspase 3 activity, increased expression of RUNX2 and OPN, type II collagen formation, ALP activity, IL-6 secretion, as well as elevated pSTAT3 phosphorylation. The differences were statistically significant compared to Si-NC group (P &lt; 0.05). Mir-149 siRNA + WP1066 inhibited pSTAT3 phosphorylation, reduced BMSCs survival, increased Caspase 3 activity, decreased RUNX2 and OPN expression, type II collagen production, ALP activity, as well as reduced IL-6 secretion. Compared with Mir-149 siRNA group, there were significant differences (P &lt; 0.05). Down-regulation of Mir-149 in BMSCs can promote BMSCs survival and osteogenic differentiation by regulating IL-6/STAT3 signaling pathway.

Targeted siRNA Nanoparticles for Mammary Carcinoma Therapy.

Non-viral, polymeric-based, siRNA nanoparticles (NPs) have been proposed as promising gene delivery systems. Encapsulating siRNA in targeted NPs could confer improved biological stability, extended half-life, enhanced permeability, effective tumor accumulation, and therapy. In this work, a peptide derived from apolipoprotein B100 (ApoB-P), the protein moiety of low-density lipoprotein, was used to target siRNA-loaded PEGylated NPs to the extracellular matrix/proteoglycans (ECM/PGs) of a mammary carcinoma tumor. siRNA against osteopontin (siOPN), a protein involved in breast cancer development and progression, was encapsulated into PEGylated poly(d,l-lactic-co-glycolic acid) (PLGA) NPs using the double emulsion solvent diffusion technique. The NPs obtained possessed desired physicochemical properties including ~200 nm size, a neutral surface charge, and high siOPN loading of ~5 µg/mg. ApoB-P-targeted NPs exhibited both enhanced binding to isolated ECM and internalization by MDA-MB-231 human mammary carcinoma cells, in comparison to non-targeted NPs. Increased accumulation of the targeted NPs was achieved in the primary mammary tumor of mice xenografted with MDA-MB-231 mammary carcinoma cells as well as in the lungs, one of the main sites affected by metastases. siOPN NPs treatment resulted in significant inhibition of tumor growth (similar bioactivity of both formulations), accompanied with significant reduction of OPN mRNA levels (~40% knockdown of mRNA levels). We demonstrated that targeted NPs possessed enhanced tumor accumulation with increased therapeutic potential in mice models of mammary carcinoma.

Physiological cyclic hydrostatic pressure induces osteogenic lineage commitment of human bone marrow stem cells: a systematic study

BackgroundPhysical loading is necessary to maintain bone tissue integrity. Loading-induced fluid shear is recognised as one of the most potent bone micromechanical cues and has been shown to direct stem cell osteogenesis. However, the effect of pressure transients, which drive fluid flow, on human bone marrow stem cell (hBMSC) osteogenesis is undetermined. Therefore, the objective of the study is to employ a systematic analysis of cyclic hydrostatic pressure (CHP) parameters predicted to occur in vivo on early hBMSC osteogenic responses and late-stage osteogenic lineage commitment.MethodshBMSC were exposed to CHP of 10 kPa, 100 kPa and 300 kPa magnitudes at frequencies of 0.5 Hz, 1 Hz and 2 Hz for 1 h, 2 h and 4 h of stimulation, and the effect on early osteogenic gene expression of COX2, RUNX2 and OPN was determined. Moreover, to decipher whether CHP can induce stem cell lineage commitment, hBMSCs were stimulated for 4 days for 2 h/day using 10 kPa, 100 kPa and 300 kPa pressures at 2 Hz frequency and cultured statically for an additional 1–2 weeks. Pressure-induced osteogenesis was quantified based on ATP release, collagen synthesis and mineral deposition.ResultsCHP elicited a positive, but variable, early osteogenic response in hBMSCs in a magnitude- and frequency-dependent manner, that is gene specific. COX2 expression elicited magnitude-dependent effects which were not present for RUNX2 or OPN mRNA expression. However, the most robust pro-osteogenic response was found at the highest magnitude (300 kPa) and frequency regimes (2 Hz). Interestingly, long-term mechanical stimulation utilising 2 Hz frequency elicited a magnitude-dependent release of ATP; however, all magnitudes promoted similar levels of collagen synthesis and significant mineral deposition, demonstrating that lineage commitment is magnitude independent. This therefore demonstrates that physiological levels of pressures, as low as 10 kPa, within the bone can drive hBMSC osteogenic lineage commitment.ConclusionOverall, these findings demonstrate an important role for cyclic hydrostatic pressure in hBMSCs and bone mechanobiology, which should be considered when studying pressure-driven fluid shear effects in hBMSCs mechanobiology. Moreover, these findings may have clinical implication in terms of bioreactor-based bone tissue engineering strategies.

Long noncoding RNA MALAT1 promotes osterix expression to regulate osteogenic differentiation by targeting miRNA-143 in human bone marrow-derived mesenchymal stem cells.

Osteogenic differentiation of human bone marrow-derived mesenchymal stem cells (hBMSCs) is essential for the human bone formation, and emerging evidence shows that long non-coding RNAs (lncRNAs) play important roles in hBMSC osteogenic differentiation. MALAT1 is often regarded as a tumor-related lncRNA, but its function in mesenchymal stem cell differentiation remains to be defined. In this study, we aimed to investigate whether MALAT1 regulates Osterix (Osx) expression by sponging miR-143 to promote hBMSC osteogenic differentiation. Firstly, we found that the expression of MALAT1 was much lower in hBMSCs from osteoporosis patients and miR-143 was contrarily higher. In addition, MALAT1 expression increased, and miR-143 decreased when hBMSCs were treated with osteogenic induction. Then, we used short hairpin RNAs to knockdown MALAT1, and the results showed that hBMSC osteogenic differentiation decreased significantly, indicating that MALAT1 is a positive regulator of osteogenic differentiation in hBMSCs. Furthermore, by luciferase assays, we found that MALAT1 could directly bind to miR-143 and negatively regulate its expression. Similarly, miR-143 could directly bind to the target site on the Osx 3'-UTR and then inhibit Osx expression. Knockdown of MALAT1 decreased Osx expression, and co-transfection of miR-143 inhibitor could rescue Osx mRNA expression. While Osx expression was increased in MALAT1-overexpressing hBMSCs, it was reversed by the miR-143 mimics. Moreover, Osx silencing decreased ALP, OCN, and OPN mRNA expression induced by the miR-143 inhibitor. Altogether, our findings suggest that MALAT1 acts to regulate Osx expression through targeting miR-143; thus, it is considered as a positive regulator in hBMSC osteogenic differentiation.

Overexpression of miR-182 inhibits ossification of ligamentum flavum cells by targeting NAMPT

Ossification of the ligamentum flavum (OLF) is a debilitating disease resulting from the development of ectopic bone formation, which leads to the compression of the spinal cord. Nicotinamide phosphoribosyltransferase (NAMPT) was found to be upregulated and microRNA-182 (miR-182) was downregulated in OLF tissue. We investigated the effects of NAMPT and miR-182 expression in OLF cells and the influence on proteins associated with osteogenic differentiation. MiR-182 overexpression inhibited NAMPT, RUNX2, OCN and OPN mRNA and protein expression in OLF cells. Alkaline phosphatase (ALP) assay and Alizarin red staining confirmed reduced levels of osteogenic differentiation and mineralized nodule formation. Knockdown of NAMPT and the NAMPT inhibitor FK866 also inhibits RUNX2, OCN and OPN mRNA expression and protein levels, whereas overexpression of NAMPT promotes the expression of RUNX2, OCN and OPN and the generation of bone nodules. Dual-luciferase reporter assays revealed that miR-182 directly targets NAMPT and downregulates its expression. Transfection of OLF cells with miR-182 downregulated NAMPT and suppressed the regulation of RUNX2, OCN, and OPN by NAMPT overexpression. Overall, these data demonstrate that miR-182 suppresses OLF by downregulating NAMPT.

Effects of small molecules water that may retard kidney stone formation.

Water intake is important for preventing kidney stones. Small molecules water is a more active restructured water under magnetic field. Here, we studied the relation between small molecules water and crystal formation in the rat kidney. The small molecules water was prepared by a water machine providing a 0.8-T magnetic field. Calcium oxalate crystals were induced by 0.75% EG (ethylene glycol) in male Sprague-Dawley rats by drinking small molecules water and plain water for up to 6weeks, respectively. Urinary ions were assayed. Osteopontin mRNA expression and urinary LDH were detected. Crystals were observed using a light microscope and a polarizing microscope. A significantly reduced urinary calcium and phosphorus excretion occurred in the 2nd and the 4th week after treatment of small molecules water. Crystals were initially detected in 40% of the experimental rats in the small molecules water group at the 6th week, later than the control group in which crystals were detected in 60% of rats at the 4th week. After 6weeks of treatment, crystals were observed to form in renal cortex, medulla and papilla in the control group, whereas only to form in renal medulla and papilla in the small molecules water group. OPN mRNA expression significantly increased earlier in the 2nd week after treatment of the small molecules water compared to the control (P=0.016). Small molecules water may retard crystal formation, reduce urinary calcium and phosphorus excretion and promote earlier OPN mRNA expression in the rat kidney.

Serum osteopontin: a biomarker of disease activity and predictor of relapsing course in patients with giant cell arteritis. Potential clinical usefulness in tocilizumab-treated patients

BackgroundOsteopontin (OPN) is a glycoprotein involved in Th1 and Th17 differentiation, tissue inflammation and remodelling. We explored the role of serum OPN (sOPN) as a biomarker in patients with giant...

Role of Reactive Oxygen Species during Low-Intensity Pulsed Ultrasound Application in MC-3 T3 E1 Pre-osteoblast Cell Culture

We evaluated the activation of mitogen-activated protein kinase (MAPK) activation through reactive oxygen species (ROS) by application of low-intensity ultrasound (LIPUS) to MC-3 T3 E1 pre-osteoblasts. The cells were subjected to one LIPUS application for either 10 or 20 min, and the control group was exposed to a sham transducer. For ROS inhibition, 10 μM diphenylene iodonium (DPI) was added to the cells an hour before LIPUS application. Samples were collected 1, 3, 6, 12 and 24 h after LIPUS application, and cells were evaluated for ROS generation, cell viability, gene expression and MAPK activation by immunoblot analyses. LIPUS caused a significant increase in ROS and cell viability in the non-DPI-treated group. Expression of RUNX2, OCN and OPN mRNA was higher in the LIPUS-treated groups at 1 h in both the DPI-treated and non-DPI-treated groups; RUNX2 and OCN mRNA levels increased at 6 h. ERK1/2 activation was increased in the LIPUS-treated groups. These results indicate that LIPUS activates MAPK by ROS generation in MC-3 T3 E1 pre-osteoblasts.

Evaluation of the prognostic value of RANK, RANKL, OPG and OPN mRNA expression in high-risk early breast cancer patients treated with anthracycline-containing adjuvant chemotherapy.

e23244Background: Prevention of bone metastases is a major issue for breast cancer patients, as it would improve quality of life in a population where long survival with metastatic disease is antic...

Preconception endurance training with voluntary exercise during pregnancy positively influences on remodeling markers in female offspring bone

Objective: In this study, we investigate the effects of preconception endurance training with or without voluntary exercise during pregnancy on indices of bone formation and resorption in female offspring bone.Methods: Twenty-four C57BL/6 female mice were randomly divided into four groups: trained in preconception period and exercised during pregnancy (TE); trained in preconception periods but unexercised during pregnancy (TC); untrained in preconception periods but exercised during pregnancy (CE); untrained and unexercised (CC). Trained dams were subjected to a protocol of moderate exercise training over a period of 4 weeks before pregnancy. Analyses were performed on the adult female offspring that did not have access to running wheels in any portion of their lives.Results: The OPG, Runx2, COLI, ALP, and OPN mRNA expression was significantly up-regulated in offspring born to dams that was trained in preconception period. However, there was no significant difference in OPG, COLI, Runx2, and ALP expression in TE and TC offspring (p > 0.05). RANKL and osteocalcin expression were significantly down-regulated in TE offspring group (p < 0.001).Conclusions: Improved physical fitness in preconception period results in significant changes in bone gene expressions of female offspring, in particular towards osteogenic responses with improved RANKL/OPG ratio.

Different response of osteoblastic cells to Mg(2+), Zn(2+) and Sr(2+) doped calcium silicate coatings.

Mg(2+), Zn(2+) and Sr(2+) substitution for Ca(2+) in plasma sprayed calcium silicate (Ca-Si) coatings have been reported to impede their degradation in physiological environment and, more importantly, to improve their biological performance. The reason for the improved biological performance is still elusive and, especially, the contribution of the dopant ions is lack of obvious and direct evidence. In this study, we aim to identify the effect of Mg(2+), Zn(2+) and Sr(2+) incorporation on the osteogenic ability of Ca-Si based coatings (Ca2MgSi2O7, Ca2ZnSi2O7 and Sr-CaSiO3) by minimizing the influence of Ca and Si ions release and surface physical properties. Similar surface morphology, crystallinity and roughness were achieved for all samples by optimizing the spray parameters. As expected, Ca and Si ions release from all the coatings showed the comparable concentration with immersing time. The response of MC3T3-E1 cells onto Mg(2+), Zn(2+) and Sr(2+) doped Ca-Si coatings were studied in terms of osteoblastic adhesion, proliferation, differentiation and mineralization. The results showed that the level of cell adhesion and proliferation increased the most on the surface of Mg-modified coating. Gene expressions of early markers of osteoblast differentiation (COL-I and ALP mRNA) were obviously improved on Zn-modified coating. Gene expressions of later markers for osteoblast differentiation (OPN and OC mRNA) and mineralized nodules formation were obviously accelerated on the surface of Sr-modified coating. Since Mg(2+), Zn(2+) and Sr(2+) play a regulatory role in different stages of osteogenesis, it may be possible to utilize this in the development of new coating materials for orthopedic application.

The influence of MicroRNA-150 in Osteoblast Matrix Mineralization.

This study investigated the influence of miR-150 expression on osteoblast matrix mineralization and its mechanisms. The mouse osteoblast cell line MC3T3-E1 was used as an in vitro model of bone formation. On the fifth day of mineralization, transfection experiments using agomiR-150, agomiR-NC, antagomiR-150 antagomiR-NC, and mock groups were set up to test the effects of miR-150 in MC3T3-E1 model. The mRNA and protein levels of OC, ALP, type I collagen, and OPN were measured by qRT-PCR and ELISA. Matrix mineralization was detected by alizarin red S (ARS) staining and flow cytometry was employed to quantify apoptosis in each group. RT-PCR and Western blot were applied to detect the expression of target gene MMP14. Our results demonstrated that the endogenous expression levels of miR-150, OC, ALP, type I collagen, and OPN in MC3T3-E1 cells increased steadily. Exogenous expressions of agomiR-150 and antagomiR-150 can significantly up-/down-regulate, respectively, the expression level of miR-150 in MC3T3-E1 cells. Compared with the mock group, higher expression levels of OC, ALP, type I collagen, and OPN mRNA were observed in the agomiR-150 group, while lower mRNA expression levels of OC, ALP, type I collagen, and OPN were found in the antagomiR-150 group. Based on these results, potential miR-150 targeted genes are discussed. Our results showed that miR-150 supports the osteoblastic phenotype related to osteoblast function and bone mineralization. Thus, miR-150 may have potential therapeutic applications in promoting bone formation in certain disease settings, such as in osteoporosis and in elderly patients.