Journal: Materials Today Bio

Article Title: Synergistic 3D-bioprinted scaffold with multi-level adaptability for vascularized bone regeneration via osteogenesis-angiogenesis coupling

doi: 10.1016/j.mtbio.2026.102837

Figure Lengend Snippet: Characterization of composite scaffold. (A) Schematic illustration of the synergistic printing process of composite scaffolds with BMSCs. (B) Macroscopic images of 3D-printed P/GG scaffolds with different filament spacing. (C) Macroscopic images of 3D-printed scaffolds with different components. (D) Macroscopic images of different views of Mg@P/M-bMSN@GG scaffold. (E) SEM images of 3D-printed scaffolds with different components, scale bar: 200 μm, 100 μm, and 10 μm. (F–H) Stress-strain curves, compressive stress and compressive modulus of 3D-printed scaffolds (n = 3). (I) Cumulative release curve of BMP-4 from B@M-bMSN (n = 3). (J) Cumulative release curves of DMOG from D@bMSN and D@M-bMSN (n = 3). (K) Cumulative release curves of Mg ions from 3D-printed Mg@P/M-bMSN@GG scaffold (n = 3). ∗P < 0.05.

Article Snippet: BMP-4 was purchased from MedChemExpress (New Jersey, USA).

Techniques:

Journal: The FASEB Journal

Article Title: Low‐Intensity Pulsed Ultrasound Promotes Osteogenesis in Porous Titanium Alloys Through miR ‐1187/ BMP4 Pathway

doi: 10.1096/fj.202403395RR

Figure Lengend Snippet: miR‐1187 directly targets BMP4. (A) The predicted binding site between miR‐1187 and BMP4 by bioinformatics analysis. (B) The luciferase activity of the BMP4‐WT and BMP4‐MUT in MC3T3‐E1 cells treated with miR‐1187 mimics or NC. (C) mRNA expression of Bmp4 was significantly downregulated in the miR‐1187 mimics‐transfected group. (D) Western blot analysis for the expression of BMP4 protein in miR‐1187 mimics and inhibitor‐transfected MC3T3‐E1 cells on Pti. (E) ELISA assay analysis for the expression of supernatant BMP4 secreted protein in miR‐1187 mimics and inhibitor‐transfected MC3T3‐E1cells on Pti. All values represent means ± SD.( n = 3). * p < 0.05, ** p < 0.01.

Article Snippet: BMP4 protein levels in the supernatant were quantified using a mouse BMP‐4 ELISA kit (EK0316, Boster, China) following the manufacturer's protocol.

Techniques: Binding Assay, Luciferase, Activity Assay, Expressing, Transfection, Western Blot, Enzyme-linked Immunosorbent Assay

Journal: The FASEB Journal

Article Title: Low‐Intensity Pulsed Ultrasound Promotes Osteogenesis in Porous Titanium Alloys Through miR ‐1187/ BMP4 Pathway

doi: 10.1096/fj.202403395RR

Figure Lengend Snippet: BMP4 positively regulates osteogenic differentiation in MC3T3‐E1 cells. (A) Level of BMP4 decreased after transfected with different siRNA‐BMP4, si‐BMP4‐785 was identified to have the highest transfection efficiency. (B) BMP4 transcript levels was determined by RT‐PCR after transfected with overexpression plasmids of BMP4. (C) qRT‐PCR analysis. Level of Col‐1 , Alp , Runx2 , and Ocn mRNA expression was significantly downregulated in BMP4 silencing. (D) qRT‐PCR analysis. Level of Col‐1 , Alp , Runx2 , and Ocn mRNA expression was significantly upregulated in BMP4 overexpression. (E) Western blotting. Expression of BMP4 protein and osteogenesis‐related proteins COL‐1, ALP, RUNX2 as well as statistical analysis after transfecting si‐BMP4. (F) Western blotting. Expression of BMP4 protein and osteogenesis‐related proteins COL‐1, ALP, RUNX2 as well as statistical analysis after transfecting pCDNA3.1‐BMP4. (G) ALP activity detection on day 7 in si‐BMP4 and pCDNA3.1‐BMP4 transfected MC3T3‐E1cells. (H) ALP staining on day 10 in si‐BMP4 and pCDNA3.1‐BMP4 transfected MC3T3‐E1cells (scale bar = 500 μm). (I) Alizarin red S staining and quantitative analysis of Alizarin Red S accumulation on day 21 in si‐BMP4 and pCDNA3.1‐BMP4 transfected MC3T3‐E1cells (scale bar = 200 μm/100 μm), the black arrow indicate the magnified area, the magnified pictures were marked within the square frame in the image. All values represent means ± SD ( n = 3). * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: BMP4 protein levels in the supernatant were quantified using a mouse BMP‐4 ELISA kit (EK0316, Boster, China) following the manufacturer's protocol.

Techniques: Transfection, Reverse Transcription Polymerase Chain Reaction, Over Expression, Quantitative RT-PCR, Expressing, Western Blot, Activity Assay, Staining

Journal: The FASEB Journal

Article Title: Low‐Intensity Pulsed Ultrasound Promotes Osteogenesis in Porous Titanium Alloys Through miR ‐1187/ BMP4 Pathway

doi: 10.1096/fj.202403395RR

Figure Lengend Snippet: LIPUS promotes osteogenic differentiation of MC3T3‐E1 through inhibiting miR‐1187 and upregulating BMP4. (A) qRT‐PCR analysis. Effect of daily LIPUS stimulation on mRNA expression of Bmp4. (B) ELISA. Supernatant BMP4 secreted by cultured MC3T3‐E1 cells was determined by ELISA assay. (C) Western blotting. BMP4 level was analyzed by western blot after 7 days LIPUS treatment. (D) Expression of osteogenesis‐related mRNA was analyzed by qRT‐PCR after si‐BMP4 or si‐bmp4 + LIPUS treatment. (E) Expression of osteogenesis‐related protein was analyzed by western blot after si‐BMP4 or si‐BMP4 + LIPUS treatment. (F) ALP activity detection on day 7 in si‐BMP4 and si‐BMP4 + LIPUS‐treated MC3T3‐E1 cells. (G) Bmp4 and osteogenesis‐related genes Col‐1, Alp, Runx2, and Ocn expression in MC3T3‐E1 cells on Pti as indicated treatment by qRT‐PCR. (H) ALP activity in MC3T3‐E1 cells on Pti on day 7 as indicated treatment. (I) Expression of BMP4 and osteogenesis‐related proteins including COL‐1, ALP, RUNX2 in MC3T3‐E1 cells on Pti as indicated treatment by Western blot analysis. All values represent means ± SD ( n = 3). Expression of osteogenesis‐related protein was analyzed by western blot (E1) and quantitative analysis (E2) after si‐BMP4, LIPUS or si‐BMP4+LIPUS treated. Expression of BMP4 and osteogenesis‐ related proteins including COL‐ 1, ALP, RUNX2 in MC3T3‐ E1 cells on Pti as indicated treatment by Western blot analysis(I)andquantitative analysis (J). * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: BMP4 protein levels in the supernatant were quantified using a mouse BMP‐4 ELISA kit (EK0316, Boster, China) following the manufacturer's protocol.

Techniques: Quantitative RT-PCR, Expressing, Enzyme-linked Immunosorbent Assay, Cell Culture, Western Blot, Activity Assay

Journal: The FASEB Journal

Article Title: Low‐Intensity Pulsed Ultrasound Promotes Osteogenesis in Porous Titanium Alloys Through miR ‐1187/ BMP4 Pathway

doi: 10.1096/fj.202403395RR

Figure Lengend Snippet: LIPUS regulates bone formation in vivo. (A) Study plan of the vitro study. (B) qRT‐PCR analysis of mRNA expression of Bmp4 as well as other osteogenesis‐related gene in the de novo bone of different groups. B1: MRNA expression of Bmp4; B2, B3: Osteogenesis‐related gene including Col‐1, Alp, Runx2, and Ocn at 4 weeks and 8 weeks respectively. (C) Micro‐CT analysis. C1: 3D reconstruction of the bone defect area at week 4 and 8. More bone ingrowth is observed as time increases in each group. The white part is the scaffold and the red part is the bone tissue. C2: The POF values for the Pti at 4 and 8 weeks. The POF values differ significantly between the LIPUS and control group, between the si‐BMP4 and control group, as well as between the si‐BMP4 and si‐BMP4 + LIPUS group. (D) D1: The representative merged images of fluorescent double labeling of calcein(green color) and xylenol orange(orange color). The red square indicate the magnified area, the magnified pictures were marked within the white square frame in the image. (scale bar = 200 μm/100 μm). D2: The MAR of the four groups at 4 and 8 weeks. All values represent means ± SD ( n = 3). * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: BMP4 protein levels in the supernatant were quantified using a mouse BMP‐4 ELISA kit (EK0316, Boster, China) following the manufacturer's protocol.

Techniques: In Vivo, Quantitative RT-PCR, Expressing, Micro-CT, Control, Labeling

Journal: The FASEB Journal

Article Title: Low‐Intensity Pulsed Ultrasound Promotes Osteogenesis in Porous Titanium Alloys Through miR ‐1187/ BMP4 Pathway

doi: 10.1096/fj.202403395RR

Figure Lengend Snippet: The schematic of LIPUS promotes osteogenesis of porous titanium alloys through inhibiting miR‐1187 and upregulating BMP4.

Article Snippet: BMP4 protein levels in the supernatant were quantified using a mouse BMP‐4 ELISA kit (EK0316, Boster, China) following the manufacturer's protocol.

Techniques:

Journal: Cell Proliferation

Article Title: Extracts from plastrum testudinis promote proliferation of rat bone‐marrow‐derived mesenchymal stem cells

doi: 10.1111/j.1365-2184.2007.00431.x

Figure Lengend Snippet: Effect of PTE on the BMP4 mRNA and BMP4 expression of MSCs in vitro. (a) Photomicrograph showing positive signal of hybridization is shown as deep brown colour within the cytoplasm in situ hybridization for BMP4 mRNA (arrow, the position where brown colour was expressed at cell cytoplasm was positive, and with haematoxylin counter stain, original magnification ×200) and BMP4‐positive cell cytoplasm were in green (white arrow, the position where green colour was expressed at cell cytoplasm was positive, the position where green colour was not expressed at cell cytoplasm was negative, and with PI counter stain, original magnification ×200), MSCs in control group showed a little BMP4 mRNA and BMP4‐positive staining, PTE treated MSCs show more strong BMP4 mRNA and BMP4‐positive staining. (b) Comparison of the percentages of BMP4 mRNA and BMP4‐positive cell among the four experimental groups after PTE treatment of 3 days. (c) Representative real‐time reverse transcription‐polymerase chain reaction demonstrating an increase in BMP4 mRNA isolated from MSC in response to a different dose of PTE (30 and 300 µg/ml culture media) for 1, 3 and 5 days. (d) Representative ELISA for BMP4 of MSCs cultured increased with increasing time and dose of PTE. *P < 0.05; **P < 0.01 compared to control.

Article Snippet: CD44, probes of BMP4 mRNA, digoxigenin‐labelled probe detection kit, and diaminobenzidine were purchased from Wuhan Boster Biological Technology Co., Ltd. (Wuhan, China); chemicals such as dimethyl sulphoxide (DMSO) and other reagents were also obtained from Sigma.

Techniques: Expressing, In Vitro, Hybridization, In Situ Hybridization, Staining, Control, Comparison, Reverse Transcription, Polymerase Chain Reaction, Isolation, Enzyme-linked Immunosorbent Assay, Cell Culture

Journal: Molecular and Cellular Biology

Article Title: Smad7 Regulates the Adult Neural Stem/Progenitor Cell Pool in a Transforming Growth Factor ?- and Bone Morphogenetic Protein-Independent Manner ▿

doi: 10.1128/MCB.00434-09

Figure Lengend Snippet: In vivo fate determination of newborn cells. (A) Newborn neurons in the dentate gyrus were analyzed by staining for the marker doublecortin (dcx). (B) Quantitative stereological analysis. Data are presented as the number of positive cells per volume (mean ± standard deviation). Statistical analysis was performed using Mann-Whitney's nonparametric test; P = 0.152. (C and D) Mice were injected with BrdU over 4 days and sacrificed 30 days later. Brain sections were double stained using antibodies for BrdU and for the cell-type-specific markers NeuN (neural cells [C]) and GFAP (astrocytes [D]). The dentate gyrus (HC) and the olfactory bulb (OF) were analyzed for the percentage of double-labeled cells (right-hand panel, depicted as mean ± standard deviation). P = 0.029 for BrdU-NeuN double staining in the hippocampus; ns, not significant. (E and F) In vitro differentiation of cultured NPCs derived from wild-type and S7ΔEx1 mice. Cells were plated on laminin-polyornithine-coated slides and kept under differentiation conditions for 7 days in the presence or absence of BMP (BMP2 and -4, 10 ng/ml each) and stained for the differentiation markers GFAP (astrocytes) and Mab2 (neurons),. The results of the quantification for neurons and for astrocytes are shown in panels E and F, respectively. Values are given as percentages of positive cells (mean ± standard deviation). *, P = 0.0236 for comparison of wild-type and S7ΔEx1 cells treated with BMP.

Article Snippet: Growth factors tested were TGF-β1 and -β2 (Peprotech Inc.), BMP2 (ImmunoTools Inc.), and BMP7 (kindly provided by K. Sampath, Curis Inc., Boston, MA); inhibitors used were TβRI-kinase inhibitors LY2109761 (Eli Lilly) and SB431542 (Sigma), recombinant noggin (BMP antagonist, Peprotech), p38 inhibitor SB203580 (Calbiochem, CA), and EGF receptor (EGFR) inhibitor AG1478 (Sigma).

Techniques: In Vivo, Staining, Marker, Standard Deviation, MANN-WHITNEY, Injection, Labeling, Double Staining, In Vitro, Cell Culture, Derivative Assay, Comparison