Design and synthesis of 3-arylbenzopyran based non-steroidal vitamin-D3mimics as osteogenic agents

MedChemComm ◽  
2016 ◽  
Vol 7 (12) ◽  
pp. 2381-2394 ◽  
Author(s):  
Mohd. Imran Ahmad ◽  
Dushyant Singh Raghuvanshi ◽  
Sarita Singh ◽  
Aijaz A. John ◽  
Ravi Prakash ◽  
...  
Keyword(s):  
Osteoblast Differentiation ◽  
Osteoblast Cells ◽  
Design And Synthesis ◽  
Calvarial Osteoblast

27benhanced osteoblast differentiation at 1 pM in mouse calvarial osteoblast cells without inherent toxicity.

scholarly journals Effects of Farnesyl Pyrophosphate Accumulation on Calvarial Osteoblast Differentiation

Endocrinology ◽  
2011 ◽  
Vol 152 (8) ◽  
pp. 3113-3122 ◽  
Author(s):  
Megan M. Weivoda ◽  
Raymond J. Hohl
Keyword(s):  
Bone Formation ◽  
Osteoblast Differentiation ◽  
Biosynthetic Pathway ◽  
Squalene Synthase ◽  
Farnesyl Pyrophosphate ◽  
Matrix Mineralization ◽  
Lower Serum Cholesterol ◽  
Calvarial Osteoblast ◽  
Isoprenoid Biosynthetic Pathway

Statins, drugs commonly used to lower serum cholesterol, have been shown to stimulate osteoblast differentiation and bone formation. Statins inhibit 3-hydroxy-3-methylglutaryl (HMG)-coenzyme A reductase (HMGCR), the first step of the isoprenoid biosynthetic pathway, leading to the depletion of the isoprenoids farnesyl pyrophosphate (FPP) and geranylgeranyl pyrophosphate (GGPP). The effects of statins on bone have previously been attributed to the depletion of GGPP, because the addition of exogenous GGPP prevented statin-stimulated osteoblast differentiation in vitro. However, in a recent report, we demonstrated that the specific depletion of GGPP did not stimulate but, in fact, inhibited osteoblast differentiation. This led us to hypothesize that isoprenoids upstream of GGPP play a role in the regulation of osteoblast differentiation. We demonstrate here that the expression of HMGCR and FPP synthase decreased during primary calvarial osteoblast differentiation, correlating with decreased FPP and GGPP levels during differentiation. Zaragozic acid (ZGA) inhibits the isoprenoid biosynthetic pathway enzyme squalene synthase, leading to an accumulation of the squalene synthase substrate FPP. ZGA treatment of calvarial osteoblasts led to a significant increase in intracellular FPP and resulted in inhibition of osteoblast differentiation as measured by osteoblastic gene expression, alkaline phosphatase activity, and matrix mineralization. Simultaneous HMGCR inhibition prevented the accumulation of FPP and restored osteoblast differentiation. In contrast, specifically inhibiting GGPPS to lower the ZGA-induced increase in GGPP did not restore osteoblast differentiation. The specificity of HMGCR inhibition to restore osteoblast differentiation of ZGA-treated cultures through the reduction in isoprenoid accumulation was confirmed with the addition of exogenous mevalonate. Similar to ZGA treatment, exogenous FPP inhibited the mineralization of primary calvarial osteoblasts. Interestingly, the effects of FPP accumulation on osteoblasts were found to be independent of protein farnesylation. Our findings are the first to demonstrate that the accumulation of FPP impairs osteoblast differentiation and suggests that the depletion of this isoprenoid may be necessary for normal and statin-induced bone formation.

Evaluation of a polyelectrolyte complex ­(PEC) composed of chitin derivatives ­and chitosan, which promotes the rat ­calvarial osteoblast differentiation

2001 ◽  
Vol 13 (1) ◽  
pp. 46-53 ◽  
Author(s):  
Takaichi Hamano ◽  
Daido Chiba ◽  
Katsuhiro Nakatsuka ◽  
Misao Nagahata ◽  
Akira Teramoto ◽  
...  
Keyword(s):  
Osteoblast Differentiation ◽  
Polyelectrolyte Complex ◽  
Calvarial Osteoblast ◽  
Chitin Derivatives

scholarly journals Porphyromonas gingivalis Lipids Inhibit Osteoblastic Differentiation and Function

2010 ◽  
Vol 78 (9) ◽  
pp. 3726-3735 ◽  
Author(s):  
Yu-Hsiung Wang ◽  
Jin Jiang ◽  
Qiang Zhu ◽  
Amer Z. AlAnezi ◽  
Robert B. Clark ◽  
...  
Keyword(s):  
Gene Expression ◽  
Porphyromonas Gingivalis ◽  
Osteoblast Differentiation ◽  
Marker Genes ◽  
Nodule Formation ◽  
Dependent Manner ◽  
Bone Homeostasis ◽  
Total Lipids ◽  
And Function ◽  
Calvarial Osteoblast

ABSTRACT Porphyromonas gingivalis produces unusual sphingolipids that are known to promote inflammatory reactions in gingival fibroblasts and Toll-like receptor 2 (TLR2)-dependent secretion of interleukin-6 from dendritic cells. The aim of the present study was to examine whether P. gingivalis lipids inhibit osteoblastic function. Total lipids from P. gingivalis and two fractions, phosphoglycerol dihydroceramides and phosphoethanolamine dihydroceramides, were prepared free of lipid A. Primary calvarial osteoblast cultures derived from 5- to 7-day-old CD-1 mice were used to examine the effects of P. gingivalis lipids on mineralized nodule formation, cell viability, apoptosis, cell proliferation, and gene expression. P. gingivalis lipids inhibited osteoblast differentiation and fluorescence expression of pOBCol2.3GFP in a concentration-dependent manner. However, P. gingivalis lipids did not significantly alter osteoblast proliferation, viability, or apoptosis. When administered during specific intervals of osteoblast growth, P. gingivalis total lipids demonstrated inhibitory effects on osteoblast differentiation only after the proliferation stage of culture. Reverse transcription-PCR confirmed the downregulation of osteoblast marker genes, including Runx2, ALP, OC, BSP, OPG, and DMP-1, with concurrent upregulation of RANKL, tumor necrosis factor alpha, and MMP-3 genes. P. gingivalis total lipids and lipid fractions inhibited calvarial osteoblast gene expression and function in vivo, as determined by the loss of expression of another osteoblast differentiation reporter, pOBCol3.6GFPcyan, and reduced uptake of Alizarin complexone stain. Finally, lipid inhibition of mineral nodule formation in vitro was dependent on TLR2 expression. Our results indicate that inhibition of osteoblast function and gene expression by P. gingivalis lipids represents a novel mechanism for altering alveolar bone homeostasis at periodontal disease sites.

Proteasome Inhibitor, Bortezomib Induces Mesenchymal Stem Cells toward Osteoblast Differentiation through Wnt-Independent Activation of Beta-catenin/TCF Signaling

Blood ◽  
2008 ◽  
Vol 112 (11) ◽  
pp. 644-644
Author(s):  
Ya-Wei Qiang ◽  
Bo Hu ◽  
Yu Chen ◽  
Ying Zhong ◽  
Bingyin Shi ◽  
...  
Keyword(s):  
Cell Lines ◽  
Proteasome Inhibitor ◽  
Osteoblast Differentiation ◽  
Progenitor Cell ◽  
Transcriptional Activity ◽  
Beta Catenin ◽  
Osteoblast Cells ◽  
Osteoblast Progenitor ◽  
Bortezomib Treatment ◽  
Msc Differentiation

Abstract In multiple myeloma (MM), the proteasome inhibitor bortezomib induces mesenchymal stem cells (MSC) toward osteoblast differentiation. However, it is full unclear about the mechanism(s) underlying bortezomib in this process. Wnt/beta-catenin pathway plays a pivotal role in osteoblast differenciation and bone development. We have demonstrated that inhibition of Wnt/beta-catenin signaling by MM-derived Dkk1 suppresses osteoblast progenitor cell differentiation into osteoblasts (Qiang et al, Bone 2008) and deregulate RANKL and OPG expression in osteoblast cells (Qiang et al Blood 2008a). Increase Wnt signaling by overexpression of Wnt3a in myeloma cells diminished MM-trigged bone lesion in mouse model (Qiang et al Blood 2008b). In the present study we revealed that bortezomib promotes MSC differentiation into osteoblast cells via Wnt-independent activation of beta-catenin/TCF signaling. E-cadherin pull-down assay and subsequently immunoblotting analysis demonstrated that bortezomib induced increases in both free and active forms of beta-catenin protein in cytoplasm and nuclear in bell-shaped dose- and time-dependent manner in mouse and human osteoblast progenitor cell lines including C2C12, C3H10T1/2, Saos-2 and MG63. Similar results were illustrated in primary human 2 cases of normal MSC and MSC from 8 cases of MM pateints. Bortezomib induced increase in ubiquitinated beta-catenin was evidenced by obvious seen slow migration bands of beta-catenin protein in SDS-PAGE gel analysis indicating that bortezomib increased beta-catenin protein by modification of proteasome-mediated degradation of beta-catenin. Increase in cytoplasm and nuclear beta-catenin protein response to bortezomib treatment in the osteoblast cell lines and 4 cases MM derived MSC was further confirmed by immunofluorescent analysis. RT-PCT analysis of TCF family revealed that abundant TCF1 and TCF4 mRNA were expressed in all tested cell lines and in a primary normal MSC, and MM-derived MSC. Bortezomib treatment also resulted in TCF transcriptional activity in bell-shaped, dose-dependent pattern as determined by luciferase activity in these cells transfected with TOPflash plasmid DNAs. Maximal responses to bortezomib were seen at 12.5 nM for both C2C12 (p<0.001) and MG63 (P<0.01), 25 nM for C3H10T1/2 (p<0.001) (p<0.00001) compared with non-stimulation control. These results suggest that transcriptional activation was a downstream effect of bortezomib in osteoblast progenitor cell lines and MM derived MSC. Bortezomib induced increases in beta-catenin protein and TCF transcriptional activity were independent of modification expression of extracellular 19 members of Wnt family ligands, 10 members of Frizzled receptor family, LRP5/6 co-receptors, and antagonists of 4 members of Dkk and sFRP family, respectively, as determined by RT-PCR analysis. Bortezomib did not increase intracellular Dvl-3 proteins, a downstream target of Wnt pathway. Lithium chloride, an inhibitor of GSK3beta did not synergized bortezomib induced increases in beta-catenin protein or TCF transcriptional activity indicating that bortezomib active beta-catenin/TCF signaling independent of activity of GSK3beta. Blocking the catenin/TCF signaling by expressing dominant–activation of beta- negative TFC attenuated bortezomib-induced matrix mineralization indicating that bortezomib induced MSC differentiation into osteoblast through activation of beta-catenin/TCF signaling. Data from experiments in comparison with biological effect of bortezomib with Wnt3a demonstrated that bortezomib did not have effect on OPG and RANKL in these cells, while Wnt3a induces OPG mRNA and protein, but inhibited RANKL expression indicating that bortezomib may have not effect on osteoclastogensis. These results provide insights into a clinically relevant mechanism of action of bortezomib and as such a rationale for its use in the treatment of diseases related to suppression of Wnt/beta-catenin/TCF signaling.

scholarly journals Activation of p38 Mitogen-Activated Protein Kinase Is Required for Osteoblast Differentiation

Endocrinology ◽  
2003 ◽  
Vol 144 (5) ◽  
pp. 2068-2074 ◽  
Author(s):  
Yuanyu Hu ◽  
Emily Chan ◽  
Sherry X. Wang ◽  
Baojie Li
Keyword(s):  
Bone Marrow ◽  
P38 Mapk ◽  
Osteoblast Differentiation ◽  
Specific Inhibitor ◽  
Dominant Negative ◽  
Mitogen Activated Protein Kinase ◽  
Bone Morphogenetic Protein 2 ◽  
Differentiation Markers ◽  
Mineral Deposition ◽  
Calvarial Osteoblast

p38 MAPK is a conserved subfamily of MAPKs involved in inflammatory response, stress response, cell growth and survival, as well as differentiation of a variety of cell types. In this report we demonstrated that p38 MAPK played an important role in osteoblast differentiation using primary calvarial osteoblast, bone marrow osteoprecursor culture, and a murine cell line, MC3T3-E1. We found that p38 MAPK was activated as calvarial osteoblast differentiates along with extracellular signal-regulated kinases (ERKs). When p38 MAPK is inhibited with a specific inhibitor, the expression of differentiation markers, such as alkaline phosphatase and mineral deposition, were significantly reduced. MC3T3-E1 cells expressing dominant negative p38 MAPK also displayed signs of delay in ALP and mineral deposition. Differentiation of the bone marrow osteoprecursors was also impeded by the p38 MAPK inhibitor, justified by the same markers. Yet the inhibitory effects observed in calvarial osteoblasts and bone marrow osteoprogenitor cells could be partially prevailed by bone morphogenetic protein-2. Inhibition of ERKs with a specific drug did not significantly affect osteoblast differentiation even though ERK1/2 were also activated during osteoblast differentiation. These results taken together indicate that p38 MAPK, but not ERKs, is necessary for osteoblast differentiation.

scholarly journals Osteogenic Activity of Lupeol Isolated from Clinacanthus nutans Lindau: Activity and Mode of Action

2021 ◽  
Vol 2021 ◽  
pp. 1-8
Author(s):  
Minh T. H. Nguyen ◽  
Quang V. Ngo ◽  
Huyen T. T. Nguyen ◽  
Quan M. Pham ◽  
Trang H. Dinh ◽  
...  
Keyword(s):  
Bone Regeneration ◽  
Signaling Pathway ◽  
Osteoblast Differentiation ◽  
Mapk Signaling ◽  
Mapk Signaling Pathway ◽  
Marker Genes ◽  
Osteoblast Cells ◽  
Traditional Use ◽  
Osteogenic Activity ◽  
Clinacanthus Nutans

Clinacanthus nutans Lindau has been traditionally used for healing of bone fragility, but the mechanism of actions has not been clarified yet. In this study, the bone regeneration activity of lupeol derived from C. nutans was assessed using an in vitro model of osteoblast cells MC3T3-E1. The finding revealed that the compound was not significantly toxic to osteoblast cells at concentration of ≤40 μg/mL. Lupeol demonstrated the osteogenic activity through enhancement of alkaline phosphatase (ALP) of osteoblast cells up to 31.2%, 21%, and 12% at concentrations of 5, 10, and 20 µg/mL, respectively ( p  < 0.05). Besides, the mineralization activity was increased up to 170, 230, 185, and 117% at concentration of 5, 10, 20, and 40 μg/mL, respectively ( p  < 0.05). The marker genes related to osteoblast differentiation evaluated on the expression level in the presence of lupeol, including collagen I (col 1), osteopontin (opn), osterix (osx), and runx2, showed upregulated expression in all the test genes ( p  < 0.05). The Western blot analysis demonstrated a clear effect of lupeol on expression of p38/p-p38, and ERK/p-ERK proteins involved in the MAPK signaling pathway. Thus, lupeol isolated from C. nutans exhibited the osteogenic activity by enhancing expression of important markers of osteogenesis, as well as affected the MAPK signaling pathway relating to osteoblast differentiation. This is the first report on the detailed mechanism of action of lupeol on bone regeneration and also explains for the traditional use of this medicinal plant for bone healing.

Implant Surface Roughness Affects Osteoblast Gene Expression

2003 ◽  
Vol 82 (5) ◽  
pp. 372-376 ◽  
Author(s):  
G.B. Schneider ◽  
H. Perinpanayagam ◽  
M. Clegg ◽  
R. Zaharias ◽  
D. Seabold ◽  
...  
Keyword(s):  
Gene Expression ◽  
Real Time ◽  
Real Time Pcr ◽  
Osteoblast Differentiation ◽  
Tissue Culture Plastic ◽  
Implant Surface ◽  
Alizarin Red ◽  
Surface Microtopography ◽  
Expression Of Genes ◽  
Calvarial Osteoblast

The transcription factor Cbfa1 regulates osteoblast differentiation and expression of genes necessary for the development of a mineralized phenotype. The purpose of this study was to determine if Cbfa1 and BSPII gene expression are influenced by implant surface microtopography. Osteoblasts were cultured on 600-grit (grooved) or sandblasted (roughened) cpTi implant discs. Mineralization was evaluated by Alizarin-Red-S staining. Real Time PCR was used for quantitative analysis of Cbfa1 and BSPII gene expression. Enhanced mineralization was seen in osteoblasts grown on roughened implant surfaces relative to tissue culture plastic. Real Time PCR showed significant (P < 0.05) increases in Cbfa1 gene expression in cells grown on roughened, as compared with grooved, implant surfaces. BSPII gene expression was also increased on rough surfaces in the UMR cells, but was reduced in the rat calvarial osteoblast cultures. These results suggest that osteoblast gene expression and mineralization are affected by roughened implant surface microtopographies during osseointegration of dental implants.

scholarly journals EctopicMsx2Overexpression Inhibits andMsx2Antisense Stimulates Calvarial Osteoblast Differentiation

1999 ◽  
Vol 209 (2) ◽  
pp. 298-307 ◽  
Author(s):  
Milan Dodig ◽  
Tade Tadic ◽  
Mark S Kronenberg ◽  
Sanja Dacic ◽  
Yi-Hsin Liu ◽  
...  
Keyword(s):  
Osteoblast Differentiation ◽  
Calvarial Osteoblast
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