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: Cell Proliferation

Article Title: The synergetic effect of bioactive molecule–loaded electrospun core‐shell fibres for reconstruction of critical‐sized calvarial bone defect—The effect of synergetic release on bone Formation

doi: 10.1111/cpr.12796

Figure Lengend Snippet: (A‐D) TEM images, (E) the degradation test result and (F) in vitro cumulative drug release profiles of electrospun fibres. Black and red represent the release profiles of DEX from PLA/PEG‐DEX and PLA/PEG‐DEX‐BMP‐2 fibres, while blue and pink represent the release profiles of BMP‐2 from PLA/PEG‐DEX‐BMP‐2 and PLA/PEG‐BMP‐2 fibres. The bars in E and F indicated the standard deviation

Article Snippet: Recombination human BMP‐2 was purchased from Bioss Biological Technology Co., LTD. ε‐CL was dried and distilled before using.

Techniques: In Vitro, Standard Deviation

Journal: Cell Proliferation

Article Title: The synergetic effect of bioactive molecule–loaded electrospun core‐shell fibres for reconstruction of critical‐sized calvarial bone defect—The effect of synergetic release on bone Formation

doi: 10.1111/cpr.12796

Figure Lengend Snippet: Representative live/dead images of BMSCs on fibres cultured at 14 d: (A) PLA/PEG, (B) PLA/PEG‐DEX, (C) PLA/PEG‐BMP‐2 and (D) PLA/PEG‐DEX‐BMP‐2. Among these, the live cells were stained green, while dead cells were stained red. (E) Percentages of live cells and (F) live cell density at 4, 7 and 14 d. At each time point, there was no significant difference among the data marked by the same letter, and significant difference existed among group marked by different letters ( P < .05, n = 5, using ANOVA analysis, the bars indicated the standard deviation). The representative images of BMSCs stained by F‐actin at 7 d of (G) PLA/PEG, (H) PLA/PEG‐DEX, (I) PLA/PEG‐BMP‐2 and (J) PLA/PEG‐DEX‐BMP‐2

Article Snippet: Recombination human BMP‐2 was purchased from Bioss Biological Technology Co., LTD. ε‐CL was dried and distilled before using.

Techniques: Cell Culture, Staining, Standard Deviation

Journal: Cell Proliferation

Article Title: The synergetic effect of bioactive molecule–loaded electrospun core‐shell fibres for reconstruction of critical‐sized calvarial bone defect—The effect of synergetic release on bone Formation

doi: 10.1111/cpr.12796

Figure Lengend Snippet: The micro‐CT images of (A) PLA/PEG, (B) PLA/PEG‐DEX, (C) PLA/PEG‐BMP‐2 and (D) PLA/PEG‐DEX‐BMP‐2 after implantation in vivo for 12 wk, the analysis of (E) new bone volume fraction and (F) bone mineral density of four groups. There was no significant difference among the data marked by the same letter, and significant difference existed among group marked by different letters ( P < .05, n = 5, using ANOVA analysis, the bars indicated the standard deviation)

Article Snippet: Recombination human BMP‐2 was purchased from Bioss Biological Technology Co., LTD. ε‐CL was dried and distilled before using.

Techniques: Micro-CT, In Vivo, Standard Deviation

Journal: Cell Proliferation

Article Title: The synergetic effect of bioactive molecule–loaded electrospun core‐shell fibres for reconstruction of critical‐sized calvarial bone defect—The effect of synergetic release on bone Formation

doi: 10.1111/cpr.12796

Figure Lengend Snippet: HE staining of PLA/PEG, PLA/PEG‐DEX, PLA/PEG‐BMP‐2 and PLA/PEG‐DEX‐BMP‐2 after implantation in vivo for 12 wk. The black arrows indicate areas of new bone

Article Snippet: Recombination human BMP‐2 was purchased from Bioss Biological Technology Co., LTD. ε‐CL was dried and distilled before using.

Techniques: Staining, In Vivo

Journal: Cell Proliferation

Article Title: The synergetic effect of bioactive molecule–loaded electrospun core‐shell fibres for reconstruction of critical‐sized calvarial bone defect—The effect of synergetic release on bone Formation

doi: 10.1111/cpr.12796

Figure Lengend Snippet: Masson's trichrome staining of PLA/PEG, PLA/PEG‐DEX, PLA/PEG‐BMP‐2 and PLA/PEG‐DEX‐BMP‐2 after implantation in vivo for 12 wk. The black arrows indicate areas of new bone

Article Snippet: Recombination human BMP‐2 was purchased from Bioss Biological Technology Co., LTD. ε‐CL was dried and distilled before using.

Techniques: Staining, In Vivo

Journal: Cell Proliferation

Article Title: The synergetic effect of bioactive molecule–loaded electrospun core‐shell fibres for reconstruction of critical‐sized calvarial bone defect—The effect of synergetic release on bone Formation

doi: 10.1111/cpr.12796

Figure Lengend Snippet: High magnification of (A) HE and (B) Masson's trichrome staining of PLA/PEG‐DEX‐BMP‐2 after implantation in vivo for 12 wk

Article Snippet: Recombination human BMP‐2 was purchased from Bioss Biological Technology Co., LTD. ε‐CL was dried and distilled before using.

Techniques: Staining, In Vivo

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: 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: Orphanet Journal of Rare Diseases

Article Title: Identification and characterization of regulatory elements in the promoter of ACVR1 , the gene mutated in Fibrodysplasia Ossificans Progressiva

doi: 10.1186/1750-1172-8-145

Figure Lengend Snippet: Effect of BMP2 on ACVR 1 promoter activity. A) HeLa and C2C12 cells were transfected with Pr-2.9 and Pr-0.072 reporter constructs and Luciferase expression was determined in the presence or absence of 100 ng/ml BMP2. Results are expressed as fold activation relative to the activity by the same promoter construct in absence of BMP2 (UN, untreated), with p < 0.05 * , p < 0.01 ** , or p < 0.001***. B) ACVR1 mRNA expression levels were detected by RT-qPCR in C2C12 cells upon treatment with BMP2. Values were normalized to the expression level of GAPDH and β - Actin genes.

Article Snippet: Human recombinant BMP2 was purchased from R&D Systems (Minneapolis, MN).

Techniques: Activity Assay, Transfection, Construct, Luciferase, Expressing, Activation Assay, Quantitative RT-PCR

Journal: British Journal of Pharmacology

Article Title: The polyphenol resveratrol promotes skeletal growth in mice through a sirtuin 1‐bone morphogenic protein 2 longevity axis

doi: 10.1111/bph.14477

Figure Lengend Snippet: eNOS and NO‐donors stimulate bone growth. eNOS mRNA expression in MC3T3 (A) and 2T3 (B) cells treated with RSV (1–100 μM) or vehicle for 24 h ( n = 5) determined by real time PCR. Nitrite (NO 2 − ) levels (C) or total eNOS protein (D) from 2T3 or MC3T3 cells treated with RSV (1–100 μM) or vehicle ( n = 5) for 48 h, determined by colorimetric assay. ALP levels from primary osteoblasts treated with NO‐donor (NOC22, 0.1–10 μM) or vehicle for 4 days ( n = 5) normalized to total cell protein (E). Calvariae from newborn mice cultured with NO‐donor (NOC22, 0.1–1 μM) or vehicle control for 4 days ( n = 5) processed for histology and H&E staining (F). mRNA expression of osteoblast marker genes Runx2, osteocalcin (OCN) and BMP2 determined in 2T3 cells treated with NOC22 (1.0 μM) or vehicle for 24 h ( n = 5) by real time PCR (G). BMP2 protein levels assessed in conditioned media from NO‐donor‐treated osteoblasts (NOC22 or SNP, 3–200 μM, 48 h, n = 5), by ELISA (with rhBMP2 as standard) (H). The μCT analysis of dissected tibia from homozygous eNOS knockout ( −/− ) or wild‐type ( +/+ ) control mice (4 month, n = 10) (I) and trabecular bone volume (J) and BMD (K) analysis (* P < 0.05 vs. vehicle; * P < 0.01 vs. wild‐type control animals).

Article Snippet: Recombinant human BMP2 (hBMP2; R&D Systems Inc.) was used as a standard.

Techniques: Expressing, Real-time Polymerase Chain Reaction, Colorimetric Assay, Cell Culture, Control, Staining, Marker, Enzyme-linked Immunosorbent Assay, Knock-Out

Journal: British Journal of Pharmacology

Article Title: The polyphenol resveratrol promotes skeletal growth in mice through a sirtuin 1‐bone morphogenic protein 2 longevity axis

doi: 10.1111/bph.14477

Figure Lengend Snippet: The eNOS‐SIRT1 axis is necessary for the pro‐osteogenic effects of RSV. Different effects on ALP levels (A) BMP2 gene expression (qPCR, B) and promoter activity (C) in primary osteoblasts from eNOS knockout ( −/− ) or wild‐type ( +/+ ) control mice ( n = 5) treated with RSV (5 μM) or vehicle (24 h). RSV‐induced ALP levels (5 μM) in the presence of the BMP inhibitor noggin (500 ng·mL −1 , 48 h, n = 5), normalized to total protein (D). SIRT1 gene expression in 2T3 osteoblasts following RSV (1–100 μM) treatment, quantified by real time PCR (E), along with eNOS (F) and BMP2 (G) mRNA levels transfected with SIRT1 siRNA or scramble control ( n = 5) with and without RSV treatment (5 μM). RSV‐induced ALP levels (5 μM) in the presence of SIRT1 (or scrambled control) siRNA (H) ( n = 5) (* P < 0.05 vs. vehicle treated WT/scrambled ctrl; # P < 0.001 vs. RSV‐treated WT/scrambled ctrl).

Article Snippet: Recombinant human BMP2 (hBMP2; R&D Systems Inc.) was used as a standard.

Techniques: Gene Expression, Activity Assay, Knock-Out, Control, Real-time Polymerase Chain Reaction, Transfection

Journal: British Journal of Pharmacology

Article Title: The polyphenol resveratrol promotes skeletal growth in mice through a sirtuin 1‐bone morphogenic protein 2 longevity axis

doi: 10.1111/bph.14477

Figure Lengend Snippet: Ageing decreases bone volume and eNOS‐BMP2 expression. μCT analysis of tibia of young (3 month) or aged (12 month) mice ( n = 10) (A) determining BV/TV (B) and BMD (C). Gene expression changes (real time PCR) of eNOS (D), BMP2 (E) in ageing long bones ( n = 10). Proposed mechanism of RSV action within osteoblasts (F) (* P < 0.05).

Article Snippet: Recombinant human BMP2 (hBMP2; R&D Systems Inc.) was used as a standard.

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

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