Journal: Experimental and Therapeutic Medicine

Article Title: Triptolide inhibits the function of TNF-α in osteoblast differentiation by inhibiting the NF-κB signaling pathway

doi: 10.3892/etm.2017.4749

Figure Lengend Snippet: TNF-α significantly inhibited the BMP-2-induced osteoblast differentiation. (A) Effect of BMP-2 on osteogenic activity. C2C12 cells were treated with 200 ng/ml BMP-2 for 3 days and the ALP activity was assessed. **P<0.01 vs. Ctrl group. (B) Effect of TNF-α on osteoblast activity. C2C12 cells were treated with 5 ng/ml TNF-α for 3 days and the ALP activity was determined. *P<0.05 vs. Ctrl group. (C) TNF-α inhibited the effect of BMP-2 on osteogenic activity. C2C12 cells were treated with 200 ng/ml BMP-2 or 200 ng/ml BMP + 5 ng/ml TNF-α for 3 days and the ALP activity was determined. ##P<0.01 vs. Ctrl group, **P<0.01 vs. BMP-2 group. TNF, tumor necrosis factor; BMP, bone morphogenetic protein; ALP, alkaline phosphatase; Ctrl, control.

Article Snippet: In the present study, C2C12 cells were treated with BMP-2 (10426-H01H-10; Sinobiological, Inc., Beijing, China) in order to produce osteoblast cells.

Techniques: Activity Assay

Journal: Experimental and Therapeutic Medicine

Article Title: Triptolide inhibits the function of TNF-α in osteoblast differentiation by inhibiting the NF-κB signaling pathway

doi: 10.3892/etm.2017.4749

Figure Lengend Snippet: TNF-α inhibited BMP-2-induced osteoblast differentiation in a dose-dependent manner. (A) C2C12 cells were treated with 100, 200 or 300 ng/ml BMP-2 for 3 days and ALP activity was evaluated. **P<0.01 vs. untreated cells. (B) C2C12 cells were treated with 100 ng/ml BMP-2 + 2.5, 5.0 or 10.0 ng/ml TNF-α for 3 days and ALP activity was assessed. **P<0.01 vs. BMP-2 group (0 ng/ml TNF-α). TNF, tumor necrosis factor; BMP, bone morphogenetic protein; ALP, alkaline phosphatase.

Article Snippet: In the present study, C2C12 cells were treated with BMP-2 (10426-H01H-10; Sinobiological, Inc., Beijing, China) in order to produce osteoblast cells.

Techniques: Activity Assay

Journal: Experimental and Therapeutic Medicine

Article Title: Triptolide inhibits the function of TNF-α in osteoblast differentiation by inhibiting the NF-κB signaling pathway

doi: 10.3892/etm.2017.4749

Figure Lengend Snippet: TPL significantly attenuated TNF-α-induced inhibition of osteoblast differentiation. C2C12 cells were treated with 100 ng/ml of BMP-2 and 5 ng/ml of TNF-α with or without triptolide (4, 8, 16 ng/ml) for 3 days and ALP activity was assessed. **P<0.01, vs. BMP-2-TNF-α group and #P<0.05, ##P<0.01, vs. BMP-2 group. TPL, triptolide; TNF, tumor necrosis factor; BMP, bone morphogenetic protein; ALP, alkaline phosphatase.

Article Snippet: In the present study, C2C12 cells were treated with BMP-2 (10426-H01H-10; Sinobiological, Inc., Beijing, China) in order to produce osteoblast cells.

Techniques: Inhibition, Activity Assay

Journal: Cell Death & Disease

Article Title: MiR-26a functions oppositely in osteogenic differentiation of BMSCs and ADSCs depending on distinct activation and roles of Wnt and BMP signaling pathway

doi: 10.1038/cddis.2015.221

Figure Lengend Snippet: Wnt and BMP signaling pathways play different role in BMSC and ADSC osteogenic differentiation. ( a–c ) BMSCs were cultured in osteogenic medium with Dorsomorphin or DKK1. Then, ALP and alizarin red staining were performed separately after 7 and 14 days of induction ( a ). Alizarin red staining was quantified by spectrophotometer ( b ). Expression of Runx2 , Alp and Ocn was measured by real-time RT-PCR ( c ). ( d–f ) ALP staining ( d ), alizarin red staining ( d and e ) and real-time RT-PCR analysis ( f ) in ADSCs cultured in osteogenic medium containing Dorsomorphin or DKK1. ( g–l ) ALP staining, alizarin red staining and real-time RT-PCR were performed in BMSCs ( g–i ) and ADSCs ( j–l ) cultured in osteogenic medium containing recombinant BMP2 or Wnt3a. Results are represented as mean±S.D. * P <0.05, ** P <0.01, *** P <0.001 ( n =3)

Article Snippet: Recombinant mouse BMP2 (R&D, Minneapolis, MN, USA), recombinant mouse Wnt3a (R&D), recombinant mouse Dkk1 (PeproTech, Rocky Hill, NJ, USA) and dorsomorphin (Santa Cruz Biotechnology, Santa Cruz, TX, USA) were obtained commercially.

Techniques: Protein-Protein interactions, Cell Culture, Staining, Spectrophotometry, Expressing, Quantitative RT-PCR, Recombinant

Journal: Endocrinology

Article Title: Msx Homeobox Genes Act Downstream of BMP2 to Regulate Endometrial Decidualization in Mice and in Humans

doi: 10.1210/en.2019-00131

Figure Lengend Snippet: Msx2 is a downstream target of BMP2 signaling in the uterus during decidualization. (A) The primary cultures of mouse endometrial stromal cells (MESCs) were transduced with adenovirus expressing GFP or BMP2. The cells were lysed at different time points, as indicated. Total RNA was isolated, and real-time PCR was performed to analyze the levels of Msx2. The relative levels of gene expression were determined by setting the expression level of the GFP-treated sample at 24 h to 1.0 (n = 3). Rplp0, encoding a ribosomal protein, was used to normalize the level of RNA. *P < 0.05. (B) The nucleotide positions of the SBEs on the Msx2 promoter were analyzed by ChIP. (C) Mouse stromal cells were treated with E + P or E, P, and BMP2 (E + P + BMP2) for 90 min. ChIP, using the Smad4 antibody, was performed, as described in “Materials and Methods.” Chromatin enrichment was quantified by real-time PCR using primers flanking the potential SBE in the Msx2 promoter and also a negative control region in the ORF of Msx2. Enrichments were normalized to 1% of input DNA. The experiment was repeated twice, and representative data are shown.

Article Snippet: The next day, the cells were either treated with E + P or E + P + BMP2 (recombinant human BMP2; R&D Systems; 355-BM-010) for 90 minutes.

Techniques: Transduction, Expressing, Isolation, Real-time Polymerase Chain Reaction, Gene Expression, Negative Control

Journal: Endocrinology

Article Title: Msx Homeobox Genes Act Downstream of BMP2 to Regulate Endometrial Decidualization in Mice and in Humans

doi: 10.1210/en.2019-00131

Figure Lengend Snippet: MSX1 and MSX2 mediate BMP2-induced HESC decidualization. The primary cultures of HESCs were established as described in “Materials and Methods.” The cells were transduced with adenovirus expressing GFP or BMP2. The cells were lysed at different time points as indicated. Total RNA was isolated, and real-time PCR was performed to analyze the levels of MSX1 and MSX2. The relative levels of gene expression were determined by setting the expression level on day 0 of the GFP-treated sample at 1.0. RPLP0, encoding a ribosomal protein, was used to normalize the level of RNA. Data were collected from three independent clinical samples, which were subjected to the same experimental conditions. *P < 0.05; **P < 0.005.

Article Snippet: The next day, the cells were either treated with E + P or E + P + BMP2 (recombinant human BMP2; R&D Systems; 355-BM-010) for 90 minutes.

Techniques: Transduction, Expressing, Isolation, Real-time Polymerase Chain Reaction, Gene Expression

Journal: Frontiers in Bioengineering and Biotechnology

Article Title: An in vivo Comparison Study Between Strontium Nanoparticles and rhBMP2

doi: 10.3389/fbioe.2020.00499

Figure Lengend Snippet: Schematic representation of abiotic and in vivo sponge characterization. The collagen sponges were loaded with calcium hydroxyapatite nanoparticles (HAn), with calcium hydroxyapatite nanoparticles + BMP2 (HAn-BMP2), and a mixture of calcium hydroxyapatite nanoparticles and strontium hydroxyapatite nanoparticles (SrHAn). Sponge characterization was achieved by several physical-chemical techniques. X-ray observations and Fast green/Safranin-O histological staining were performed on tissue samples collected from the in vivo experiments. Expression of genes for osteogenesis, osteocytes and osteoclasts activity, chondrogenesis, and stem cell recruitment was also investigated. Figure created with BioRender.com .

Article Snippet: For rhBMP2 loaded-sponges, 3 μg of human recombinant BMP2 (R&D Systems) were added to the HAn suspension before the sponge absorption phase (HAn-BMP2).

Techniques: In Vivo, Staining, Expressing, Activity Assay

Journal: Frontiers in Bioengineering and Biotechnology

Article Title: An in vivo Comparison Study Between Strontium Nanoparticles and rhBMP2

doi: 10.3389/fbioe.2020.00499

Figure Lengend Snippet: Calcium, strontium and rhBMP2 release from loaded untreated sponges. ICP-OES data representation of calcium and strontium ions release from HAn, HAn-BMP2, and SrHAn loaded sponges. Indirect ELISA was performed to quantify solubilized rhBMP2. Data were collected during the 28 days in aqueous solution. Ca 2+ released from HAn sponges (A) ; Ca 2+ and Sr 2+ released from SrHAn sponges (B) ; Ca 2+ and rhBMP2 release from HAn-BMP2 sponges (C,D) N = 3 samples were measure per each time point.

Article Snippet: For rhBMP2 loaded-sponges, 3 μg of human recombinant BMP2 (R&D Systems) were added to the HAn suspension before the sponge absorption phase (HAn-BMP2).

Techniques: Indirect ELISA

Journal: Frontiers in Bioengineering and Biotechnology

Article Title: An in vivo Comparison Study Between Strontium Nanoparticles and rhBMP2

doi: 10.3389/fbioe.2020.00499

Figure Lengend Snippet: Tabular representation of the data shown in Figure 2.

Article Snippet: For rhBMP2 loaded-sponges, 3 μg of human recombinant BMP2 (R&D Systems) were added to the HAn suspension before the sponge absorption phase (HAn-BMP2).

Techniques:

Journal: Frontiers in Bioengineering and Biotechnology

Article Title: An in vivo Comparison Study Between Strontium Nanoparticles and rhBMP2

doi: 10.3389/fbioe.2020.00499

Figure Lengend Snippet: The microscopic structure (150× and 350×) of the untreated CTRL (unloaded sponges), HAn, HAn-BMP2 and SrHAn (loaded sponges) samples is shown in (A-H) . Pictures represented samples at T0 (A–D) and after 28 days (E–H) of permanence in aqueous solutions, in a controlled and humidified atmosphere (37°C, with 5% of CO 2 ). Untreated samples surfaces were also analyzed with SEM-EDS (Energy Dispersive X-Ray Spectroscopy) at 0 (I,J) and 28 days (K,L) . Images at low magnifications of the sponges were taken with SEM (I–L) . The elements phosphorus (P), calcium (Ca) and strontium (Sr) were mapped on the same pictures using different colors (P = green; Ca = red; Sr = white) and a black background. Quantification of the elements carbon (C), oxygen (O), phosphorus (P), calcium (Ca) and strontium (Sr) present of the samples surface was performed and data were reported as weight percentage in the bar charts (M,N,O,P) histograms.

Article Snippet: For rhBMP2 loaded-sponges, 3 μg of human recombinant BMP2 (R&D Systems) were added to the HAn suspension before the sponge absorption phase (HAn-BMP2).

Techniques: Spectroscopy

Journal: Frontiers in Bioengineering and Biotechnology

Article Title: An in vivo Comparison Study Between Strontium Nanoparticles and rhBMP2

doi: 10.3389/fbioe.2020.00499

Figure Lengend Snippet: X-ray images and macroscopic observation of mice implanted with loaded sponges. Images at 16 and 33 days of mice implanted with HAn loaded-sponges ( A,D,G,J,M,P , respectively), HAn-BMP2 loaded-sponges (B,E,H,K,N,Q) and SrHAn ( C,F,I, L,O,R , respectively). The implanted sponges are shown by black arrowheads. Isolated femurs are shown in panels (D–F) and (M–O) . Isolated legs including implants are shown in panels (G–I) and (P–R) .

Article Snippet: For rhBMP2 loaded-sponges, 3 μg of human recombinant BMP2 (R&D Systems) were added to the HAn suspension before the sponge absorption phase (HAn-BMP2).

Techniques: Isolation

Journal: Frontiers in Bioengineering and Biotechnology

Article Title: An in vivo Comparison Study Between Strontium Nanoparticles and rhBMP2

doi: 10.3389/fbioe.2020.00499

Figure Lengend Snippet: Representative histological images of post-implants tissues of mouse limb. Fast green/Safranin-O staining was used on post-implant tissue sections of limbs implanted with loaded sponges with HAn (A,D,G,J) , HAn-BMP2 (B,E,H,K) , or SrHAn (C,F,I,L) , for 16 and 33 days, respectively. In the images, red spots of cartilaginous tissue are indicated with red arrowheads and gray blurs of ectopic bone are highlighted with *. Black arrows and black arrowheads indicate femur bone and sponges, respectively. The circle highlights a blood vessel.

Article Snippet: For rhBMP2 loaded-sponges, 3 μg of human recombinant BMP2 (R&D Systems) were added to the HAn suspension before the sponge absorption phase (HAn-BMP2).

Techniques: Staining

Journal: Frontiers in Bioengineering and Biotechnology

Article Title: An in vivo Comparison Study Between Strontium Nanoparticles and rhBMP2

doi: 10.3389/fbioe.2020.00499

Figure Lengend Snippet: Gene expression of stem cell recruitment and chondrogenesis markers in bone and sponge post-implants. Gene expression was analyzed in femur bone (A,B,E,F) and sponge post-implant (C,D,G,H) samples at 16 (A–D) and 33 days (E–H) , respectively. Expression of chondrogenesis markers Acan, Col10A1 and Sox9 (A,C,E,G) as well as stem cell recruitment marker Nanog (B,D,F,H) has been evaluated. The graphs show the inverse of the ΔΔCt at the power of 2. Bars indicate mean values ± SEM of results from 4 experiments. Statistical significance values were calculated with one way-ANOVA, followed by Tukey's honestly significant difference test. *, significant difference against the HAn condition ( p < 0.05). #, significant difference between HAn-BMP2 and Sr conditions ( p < 0.05).

Article Snippet: For rhBMP2 loaded-sponges, 3 μg of human recombinant BMP2 (R&D Systems) were added to the HAn suspension before the sponge absorption phase (HAn-BMP2).

Techniques: Expressing, Marker

Journal: Frontiers in Bioengineering and Biotechnology

Article Title: An in vivo Comparison Study Between Strontium Nanoparticles and rhBMP2

doi: 10.3389/fbioe.2020.00499

Figure Lengend Snippet: Gene expression of osteocytes, osteoclasts homeostasis, and osteogenic markers in bone and sponge post-implants. Gene expression was evaluated in femur bone (A,B,C,G,H,I) and sponge post-implant (D,E,F,J,K,L) samples at 16 (A–F) and 33 (G–L) days, respectively. Expression of osteogenic markers Dmp1, Bglap, Ibsp, Sp7 and Runx2 (A,D,G,J) , osteocytes marker Sost (B,E,H,K) and osteoclasts relevant markers Acp5, Rankl and Ctsk (C,F,I,L) has been evaluated. The graphs show the inverse of the ΔΔCt at the power of 2. Bars indicate mean values ± SEM of results from four experiments. Statistical significance values were calculated with one way-ANOVA, followed by Tukey's honestly significant difference test. *, significant difference against the HAn condition ( p < 0.05). #, significant difference between HAn-BMP2 and SrHAn conditions ( p < 0.05).

Article Snippet: For rhBMP2 loaded-sponges, 3 μg of human recombinant BMP2 (R&D Systems) were added to the HAn suspension before the sponge absorption phase (HAn-BMP2).

Techniques: Expressing, Marker

Journal: Immunity

Article Title: Stellate Cells, Hepatocytes, and Endothelial Cells Imprint the Kupffer Cell Identity on Monocytes Colonizing the Liver Macrophage Niche

doi: 10.1016/j.immuni.2019.08.017

Figure Lengend Snippet:

Article Snippet: Recombinant Human/Mouse/Rat BMP-2 Protein , RnD Systems , Cat#355-BM-010.

Techniques: Control, Recombinant, Blocking Assay, Irradiation, Enzyme-linked Immunosorbent Assay, Microarray, Software, Microscopy

Journal:

Article Title: PROTEIN KINASE A REGULATES GDNF/RET-DEPENDENT BUT NOT GDNF/RET-INDEPENDENT URETERIC BUD OUTGROWTH FROM THE WOLFFIAN DUCT

doi: 10.1016/j.ydbio.2010.08.029

Figure Lengend Snippet: Wolffian ducts were isolated from E13 rat embryos and cultured for two days in either control media (FGF1 and GDNF) or in the added presence 5nM BMP. A) BMP2 completely inhibited UB outgrowth from the Wolffian duct. Green – E-cadherin; blue – DAPI. Scale bar = 50µm. B) PKA enzyme activity, normalized by protein concentration assay, was calculated from the slope of the linear least-squares fit of the PKA assay in each condition; the increase in PKA enzyme activity measured in those ducts exposed to BMP2 was determined to be statistically significant C) qRT-PCR confirms significant downregulation of Ret expression in the WDPKA+ although Ret levels in the WD exposed to BMP2 were not significantly different from control. D) qRT-PCR shows significant downregulation of GFRα expression in both the WDPKA+ and WDs cultured with 5nM BMP-2 in the absence of PKA inhibitor (* = p < 0.05 versus control; NS = not significant). Scale bar = 100µm.

Article Snippet: Recombinant GDNF, fibroblast growth factor (FGF)-1, FGF-7, follistatin, and bone morphogenetic protein (BMP)-2 were purchased from R&D systems (Minneapolis, MN).

Techniques: Isolation, Cell Culture, Activity Assay, Protein Concentration, Protein Kinase A Assay, Quantitative RT-PCR, Expressing