Journal: Frontiers in Microbiology

Article Title: Shengu granules ameliorate ovariectomy-induced osteoporosis by the gut-bone-immune axis

doi: 10.3389/fmicb.2024.1320500

Figure Lengend Snippet: The effect of Shengu granules on intestinal barrier, FOXP3 cells and OPG/RANKL Signaling pathway protein expression. The colon pathological sections of the Sham, OVX and SG group, the former one (magnification, 40X), the latter one (magnification, 200X) (A) . Necrosis and exfoliation of mucosal epithelial cells (brown arrow), lymphocyte infiltration (red arrow), and nuclear fragmentation (black arrow) in Sham group; Nuclei were fragmented (black arrow), and more lymphocytes were found in the lamina propria (red arrow) in OVX group. Small focal infiltration of lymphocytes (red arrows) in SG group (A) . The Immunofluorescence results of each group (magnification, 200X) (B) . Histological score (C) . Percentage of mean Immunofluorescence intensity of FOXP3 cells in the three groups (D) . Protein blot analysis, semiquantitative analysis of protein blotting results of ZO-1 and Occludin in colon biopsy, OPG and RANKL in tibia tissue (E) . The ZO-1 and Occludin protein expression in colon tissue (F,G) . The OPG and RANKL protein expression ratio (H) . The OPG and RANKL protein expression in tibia tissue (I,J) . Statistical significance was evaluated using one-way ANOVA procedure and Tukey test. Different letters represent significant differences between groups ( p < 0.05) vs. OVX group. n = 5, * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001. Data are represented as mean ± SD.

Article Snippet: Total protein samples extracted from colon tissue were incubated with ZO-1 (Servicebio Technology Ltd., China) and Occludin (Servicebio Technology Ltd., China) primary and secondary antibodies, and total protein samples extracted from femur tissue were incubated with OPG (Servicebio Technology Ltd., China) and RANKL (Servicebio Technology Ltd., China) primary and secondary antibodies.

Techniques: Expressing, Immunofluorescence

Journal: Bioactive Materials

Article Title: An electrostatic encapsulation strategy to motivate 3D-printed polyelectrolyte scaffolds for repair of osteoporotic bone defects

doi: 10.1016/j.bioactmat.2024.12.007

Figure Lengend Snippet: Histological and immunofluorescence analysis of bone defect regeneration in OBD rats treated with APS@P scaffold. A) Representative images of H&E staining depicting the defect area across different treatment groups; B) Representative immunofluorescence staining images showing RANKL expression in each group, with green fluorescence indicating RANKL-positive expression; C) Representative immunofluorescence staining images demonstrating OCN and RUNX2 expression in each group, where green fluorescence indicates OCN-positive expression and red fluorescence indicates Runx2-positive expression; D) Representative immunofluorescence staining images illustrating CD31 and Endomucin expression in each group, with green fluorescence indicating CD31-positive expression and red fluorescence indicating Endomucin-positive expression. Yellow fluorescence indicates overlap of CD31 and Endomucin, highlighting the presence of osteogenic type H vessels. E) Quantitative analysis of RANKL expression by immunofluorescence is provided for each group; F) Quantitative analysis of OCN and RUNX2 protein expression by immunofluorescence is provided for each group; G) Quantitative analysis of CD31, Endomucin, and CD31 & Endomucin overlap expression by immunofluorescence across groups. Statistical significance was defined as ∗ P < 0.05, ∗∗ P < 0.01, and ∗∗∗ P < 0.001 indicates statistical significance. Number of samples n = 6.

Article Snippet: Primary antibodies for OCN (1:100, 29697, Signalway Antibody, USA), Runx2 (1:100, ab76956, Abcam, UK), RANKL (1:100, 41789, Signalway Antibody, USA), CD31 (1:100, ab222783, Abcam, UK), and Endomucin (1:100, sc-65495, Santa cruz biotechnology, USA) were prepared in a diluent buffer.

Techniques: Immunofluorescence, Staining, Expressing, Fluorescence

Journal: Bioactive Materials

Article Title: An electrostatic encapsulation strategy to motivate 3D-printed polyelectrolyte scaffolds for repair of osteoporotic bone defects

doi: 10.1016/j.bioactmat.2024.12.007

Figure Lengend Snippet: Potential mechanism of APS@P scaffold treatment for OBD. The APS@P scaffold integrates SAB-BTL within an alginate/ε-polylysine scaffold, allowing for sustained drug release, targeted delivery, drug protection, and enhanced therapeutic efficacy. The therapeutic mechanism of APS@P in treating OBD involves several key processes: 1. Osteogenic Differentiation, APS@P promotes the osteogenic differentiation of mesenchymal stem cells (MSCs) by activating the Tph2/Wnt/β-catenin signaling axis. This activation leads to the upregulation of osteogenic biomarkers, including Alp , Ocn , Runx2 , and Osterix . 2. Angiogenesis Promotion, the scaffold enhances angiogenesis through increased expression of vascular endothelial growth factor ( Vegf ) and erythropoietin ( Epo ). Additionally, it stimulates the formation of type H vessels, marked by co-expression of CD31 and endomucin, which are crucial for bone regeneration. 3. Inhibition of Bone Resorption, APS@P suppresses RANKL-mediated bone resorption, a key process in maintaining bone mass. These combined actions create a favorable microenvironment that restores bone homeostasis and promotes bone regeneration in OBD.

Article Snippet: Primary antibodies for OCN (1:100, 29697, Signalway Antibody, USA), Runx2 (1:100, ab76956, Abcam, UK), RANKL (1:100, 41789, Signalway Antibody, USA), CD31 (1:100, ab222783, Abcam, UK), and Endomucin (1:100, sc-65495, Santa cruz biotechnology, USA) were prepared in a diluent buffer.

Techniques: Activation Assay, Expressing, Inhibition

Journal: International Journal of Molecular Sciences

Article Title: Protective Effect of Acteoside on Ovariectomy-Induced Bone Loss in Mice

doi: 10.3390/ijms20122974

Figure Lengend Snippet: Acteoside treatment on the expression levels of TRAF6 ( A ), RANKL ( B ), RANK ( C ), NFKBIA ( D ), PI3K ( E ), AKT ( F ), IKKβ ( G ), NFAT2 ( H ), and c-Fos ( I ) ( n = 3); β-actin was shown as the loading control, and quantitative data of every signal protein was descriptive as percentages of the value of control. Values were expressed as the mean ± SD. * p < 0.05, ** p < 0.01, *** p < 0.001 relative to the control group.

Article Snippet: Co. Ltd., Nanjing, China; primary antibodies including TRAF6, RANKL, RANK, NFKBIA, PI3K, AKT, IKKβ, NFAT2, c-Fos, β-actin, and secondary antibodies of horseradish peroxidase-conjugated goat anti-rabbit IgG were offered by ZSGB-BIO, Beijing, China; all other reagents were of analytical purity.

Techniques: Expressing, Control

Journal: Microorganisms

Article Title: Probiotic Propionibacterium freudenreichii MJ2 Enhances Osteoblast Differentiation and Mineralization by Increasing the OPG/RANKL Ratio

doi: 10.3390/microorganisms9040673

Figure Lengend Snippet: Primer sequences used for qPCR.

Article Snippet: Western blotting was performed with primary antibodies against the endogenous control β-actin, OPG, and RANKL (1:1000 dilution, GTX108515; Genetex) as described above.

Techniques:

Journal: Microorganisms

Article Title: Probiotic Propionibacterium freudenreichii MJ2 Enhances Osteoblast Differentiation and Mineralization by Increasing the OPG/RANKL Ratio

doi: 10.3390/microorganisms9040673

Figure Lengend Snippet: STRING analysis of protein interaction networks. Interactions of identified proteins were mapped by searching STRING database version 10.5 with a confidence cutoff of 0.4. In the resulting protein association network, proteins are presented as nodes connected by lines whose thickness represents the confidence level. Osteoblast differentiation-related genes BGLAP (OC), TNFRSF11B (OPG), SP7 (OSX), TNFSF11 (RANKL), and FN1 (fibronectin) were obtained from the qPCR data .

Article Snippet: Western blotting was performed with primary antibodies against the endogenous control β-actin, OPG, and RANKL (1:1000 dilution, GTX108515; Genetex) as described above.

Techniques:

Journal: Microorganisms

Article Title: Probiotic Propionibacterium freudenreichii MJ2 Enhances Osteoblast Differentiation and Mineralization by Increasing the OPG/RANKL Ratio

doi: 10.3390/microorganisms9040673

Figure Lengend Snippet: Effect of MJ2 and L. plantarum on the OPG/RANKL ratio and expression of osteoblast differentiation-related genes and proteins. The gene expression levels of OPG/RANKL ( A ) and expression levels of the proteins were measured by Western blotting ( B ) and quantified ( C ). Each value indicates the mean ± SEM of three independent experiments performed in triplicate. * p < 0.05, ** p < 0.01, and *** p < 0.001 compared with the OVX group.

Article Snippet: Western blotting was performed with primary antibodies against the endogenous control β-actin, OPG, and RANKL (1:1000 dilution, GTX108515; Genetex) as described above.

Techniques: Expressing, Gene Expression, Western Blot