β₂Adrenergic Receptor Activation Suppresses Bone Morphogenetic Protein (BMP)-Induced Alkaline Phosphatase Expression in Osteoblast-Like MC3T3E1 Cells

2015 ◽  
Vol 116 (6) ◽  
pp. 1144-1152 ◽  
Author(s):  
Takayuki Yamada ◽  
Yoichi Ezura ◽  
Tadayoshi Hayata ◽  
Shuichi Moriya ◽  
Jumpei Shirakawa ◽  
...  
Keyword(s):  
Alkaline Phosphatase ◽  
Bone Morphogenetic Protein ◽  
Adrenergic Receptor ◽  
Receptor Activation ◽  
Morphogenetic Protein ◽  
Alkaline Phosphatase Expression

Key role of the expression of bone morphogenetic proteins in increasing the osteogenic activity of osteoblast-like cells exposed to shock waves and seeded on bioactive glass-ceramic scaffolds for bone tissue engineering

2014 ◽  
Vol 29 (5) ◽  
pp. 728-736 ◽  
Author(s):  
Giuliana Muzio ◽  
Germana Martinasso ◽  
Francesco Baino ◽  
Roberto Frairia ◽  
Chiara Vitale-Brovarone ◽  
...  
Keyword(s):  
Shock Wave ◽  
Alkaline Phosphatase ◽  
Shock Waves ◽  
Bioactive Glass ◽  
Bone Morphogenetic Protein ◽  
Bone Morphogenetic Proteins ◽  
Glass Ceramic ◽  
Osteogenic Activity ◽  
Morphogenetic Protein

In this work, the role of shock wave-induced increase of bone morphogenetic proteins in modulating the osteogenic properties of osteoblast-like cells seeded on a bioactive scaffold was investigated using gremlin as a bone morphogenetic protein antagonist. Bone-like glass-ceramic scaffolds, based on a silicate experimental bioactive glass developed at the Politecnico di Torino, were produced by the sponge replication method and used as porous substrates for cell culture. Human MG-63 cells, exposed to shock waves and seeded on the scaffolds, were treated with gremlin every two days and analysed after 20 days for the expression of osteoblast differentiation markers. Shock waves have been shown to induce osteogenic activity mediated by increased expression of alkaline phosphatase, osteocalcin, type I collagen, BMP-4 and BMP-7. Cells exposed to shock waves plus gremlin showed increased growth in comparison with cells treated with shock waves alone and, conversely, mRNA contents of alkaline phosphatase and osteocalcin were significantly lower. Therefore, the shock wave-mediated increased expression of bone morphogenetic protein in MG-63 cells seeded on the scaffolds is essential in improving osteogenic activity; blocking bone morphogenetic protein via gremlin completely prevents the increase of alkaline phosphatase and osteocalcin. The results confirmed that the combination of glass-ceramic scaffolds and shock waves exposure could be used to significantly improve osteogenesis opening new perspectives for bone regenerative medicine.

scholarly journals Critical Regulation of Bone Morphogenetic Protein-induced Osteoblastic Differentiation by Delta1/Jagged1-activated Notch1 Signaling

2005 ◽  
Vol 280 (16) ◽  
pp. 15842-15848 ◽  
Author(s):  
Masuhiro Nobta ◽  
Tomoo Tsukazaki ◽  
Yasuaki Shibata ◽  
Chang Xin ◽  
Takeshi Moriishi ◽  
...  
Keyword(s):  
Alkaline Phosphatase ◽  
Notch Signaling ◽  
Bone Morphogenetic Protein ◽  
Phosphatase Activity ◽  
Alkaline Phosphatase Activity ◽  
Notch Pathway ◽  
Osteoblastic Differentiation ◽  
Differentiation Markers ◽  
Induced Differentiation ◽  
Morphogenetic Protein

Functional involvement of the Notch pathway in osteoblastic differentiation has been previously investigated using the truncated intracellular domain, which mimics Notch signaling by interacting with the DNA-binding protein CBF-1. However, it is unclear whether Notch ligands Delta1 and Jagged1 also induce an identical cellular response in osteoblastic differentiation. We have shown that both Delta1 and Jagged1 were expressed concomitantly with Notch1 in maturating osteoblastic cells during bone regeneration and that overexpressed and immobilized recombinant Delta1 and Jagged1 alone did not alter the differentiated state of MC3T3-E1 and C2C12 cells. However, they augmented bone morphogenetic protein-2 (BMP2)-induced alkaline phosphatase activity and the expression of several differentiation markers, except for osteocalcin, and ultimately enhanced calcified nodule andin vivoectopic bone formation of MC3T3-E1. In addition, both ligands transmitted signal through the CBF-1-dependent pathway and stimulated the expression of HES-1, a direct target of Notch pathway. To test the necessity of Notch signaling in BMP2-induced differentiation, Notch signaling was inhibited by the dominant negative extracellular domain of Notch1, specific inhibitor, or small interference RNA. These treatments decreased alkaline phosphatase activity as well as the expression of other differentiation markers and inhibited the promoter activity of Id-1, a target gene of the BMP pathway. These results indicate the functional redundancy between Delta1 and Jagged1 in osteoblastic differentiation whereby Delta1/Jagged1-activated Notch1 enhances BMP2-induced differentiation through the identical signaling pathway. Furthermore, our data also suggest that functional Notch signaling is essential not only for BMP2-induced osteoblast differentiation but also for BMP signaling itself.

scholarly journals Inhibin Suppresses and Activin Stimulates Osteoblastogenesis and Osteoclastogenesis in Murine Bone Marrow Cultures

Endocrinology ◽  
2002 ◽  
Vol 143 (1) ◽  
pp. 74-83 ◽  
Author(s):  
D. Gaddy-Kurten ◽  
J. K. Coker ◽  
E. Abe ◽  
R. L. Jilka ◽  
S. C. Manolagas
Keyword(s):  
Bone Marrow ◽  
Alkaline Phosphatase ◽  
Bone Morphogenetic Protein ◽  
Colony Formation ◽  
Tartrate Resistant Acid Phosphatase ◽  
Murine Bone Marrow ◽  
Morphogenetic Protein ◽  
Bone Marrow Cultures ◽  
Marrow Cultures

Abstract Using primary murine bone marrow cell cultures, we demonstrate that inhibin suppresses osteoblastogenesis and osteoclastogenesis. In contrast, activin supports osteoblast formation (by alkaline phosphatase-positive and mineralized colony formation); and activin also stimulates osteoclast formation (as measured by staining tartrate-resistant acid phosphatase-positive multinucleated cells). Inhibin, the activin antagonist follistatin, and the bone morphogenetic protein antagonist noggin can all suppress endogenous activin accumulation in bone marrow cultures. Associated with this decrease in activin is the loss of mineralized osteoblastic colony formation (colony forming unit-osteoblast; CFU-OB). However, exogenous activin administration, even in the presence of noggin, permits both alkaline phosphatase-positive and CFU-OB colony formation in vitro. In contrast, the stimulatory effects of locally produced activin on osteoblast and osteoclast development are not likely to be dominant over the suppressive effects of gonadally derived inhibin. The suppressive effect of inhibin is maintained in the presence of either activin or bone morphogenetic protein, suggesting the presence of a distinct inhibin-specific receptor. Taken together, the direct regulation of osteoblastogenesis and osteoclastogenesis by inhibin and activin in vitro suggest that changes in the inhibin/activin ratio detected by bone marrow cells, during the perimenopausal transition, contribute to altered cell differentiation and may be associated with the increased bone resorption observed at this time.

Soluble expression and purification of high-bioactivity recombinant human bone morphogenetic protein-2 by codon optimisation in Escherichia coli

2019 ◽  
Vol 32 (3) ◽  
pp. 153-157
Author(s):  
Wei Chen ◽  
Caiqian Zhang ◽  
Yeqing Wu ◽  
Xiuping Su
Keyword(s):  
Escherichia Coli ◽  
Alkaline Phosphatase ◽  
Bone Morphogenetic Protein ◽  
Human Bone ◽  
Bone Morphogenetic Protein 2 ◽  
E Coli ◽  
Alp Activity ◽  
Morphogenetic Protein ◽  
Human Bone Morphogenetic Protein ◽  
Recombinant Human Bone

Abstract We developed a simple method of preparing recombinant human bone morphogenetic protein-2 (rhBMP-2) with high biological activity. This rhBMP-2 was overproduced in Escherichia coli as a fusion protein with thioredoxin 6xHis-tag at its amino terminus. The cDNA fragment of human bone morphogenetic protein-2 (hBMP-2) fused to the secretion signal of alkaline phosphatase (PhoA) was expressed under T7 promoter in E. coli. After DNA sequence confirmation, the recombinant vector pETpho-bmp2 was transformed into E. coli BL21 (DE3). rhBMP-2 was produced by the recombinant strain pETpho-bmp2/BL21 (DE3) in a soluble form with an yield of 6.2 mg/L culture. Sodium Dodecyl Sulfate Polyacrylamide Gel Electrophoresis (SDS-PAGE) results showed that the molecular weight of the product was approximately 28 kD. Moreover, rhBMP-2 was secreted as a dimer with a natural structure. rhBMP-2, purified by Ni Nitrilotriacetic acid Agarose (Ni-NTA) affinity chromatography, was used to examine osteosarcoma MG-63 cells and assay the alkaline phosphatase (ALP) activity. Results showed that rhBMP-2 induced MG-63 cell differentiation. When the final concentration was 500 ng/mL, the effect was more remarkable and ALP activity reached 525% compared with that of the control group.

scholarly journals 1-Step Versus 2-Step Immobilization of Alkaline Phosphatase and Bone Morphogenetic Protein-2 onto Implant Surfaces Using Polydopamine

2013 ◽  
Vol 19 (8) ◽  
pp. 610-619 ◽  
Author(s):  
Arnold W.G. Nijhuis ◽  
Jeroen J.J.P. van den Beucken ◽  
Otto C. Boerman ◽  
John A. Jansen ◽  
Sander C.G. Leeuwenburgh
Keyword(s):  
Alkaline Phosphatase ◽  
Bone Morphogenetic Protein ◽  
Bone Morphogenetic Protein 2 ◽  
Morphogenetic Protein
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