Journal: Biomedicines

Article Title: Recombinant BMP9 Reinforces Gut Vascular Barrier in Experimental Colitis

doi: 10.3390/biomedicines14020288

Figure Lengend Snippet: Dysregulation of BMP9/ALK1 signaling and inflammation in refractory ulcerative colitis (rUC). ( A ) Heatmap depicting serum expression levels of BMP family members in healthy controls, non-rUC, and rUC patients ( n = 3 per group). Data were Z-score normalized and hierarchically clustered (red, high expression; blue, low expression). ( B ) Baseline serum levels of BMP9 and BMP10 in healthy controls ( n = 50), non-rUC patients ( n = 47), and rUC patients ( n = 48). ( C ) Spearman correlation analyses between baseline serum BMP9 levels and clinical disease activity indices, including baseline Modified Mayo Score, baseline UCEIS, post-treatment Modified Mayo Score, and post-treatment UCEIS, in patients with UC ( n = 95). ( D ) Colonic mucosal mRNA expression levels of ALK1, IL-6, TNF-α, and CCL2 in healthy controls, non-rUC, and rUC patients ( n = 4 per group). Statistical annotations for ( B , D ): (Normalized to GAPDH; Mean ± SD; Statistical significance determined by one-way ANOVA with Tukey’s post hoc test: ** p < 0.01, *** p < 0.001, ns: not significant).

Article Snippet: Immediately after seeding, HIMECs were treated with the following stimuli: recombinant human BMP9 (rhBMP9; 0–10 ng/mL), TNF-α (20 ng/mL; Novoprotein, Suzhou, China, #C008), and ML347 (150 nM; MCE, Monmouth Junction, NJ, USA, #HY-12274).

Techniques: Expressing, Activity Assay, Modification

Journal: Biomedicines

Article Title: Recombinant BMP9 Reinforces Gut Vascular Barrier in Experimental Colitis

doi: 10.3390/biomedicines14020288

Figure Lengend Snippet: BMP9 attenuates DSS-induced colitis in mice. ( A ) Schematic of experimental design: Acute colitis was induced in C57BL/6 mice by 3% DSS in drinking water for 7 days. The BMP9 treatment group received intraperitoneal injections of recombinant murine BMP9 (200 ng/day), while the DSS group and the control group received PBS ( n = 14 per group). ( B ) Serum BMP9(ng/mL) concentrations measured by ELISA. ( C ) Representative images of colons and quantitative analysis of colon length (cm). ( D ) Colonoscopy images (upper), H&E-stained colon sections (lower; scale bars = 100 μm), and histopathological scores. ( E ) (Upper) Dynamic body weight changes and (Lower) Disease Activity Index (DAI) scores. Data expressed as mean ± SD; * p < 0.05, two-way repeated measures ANOVA. ( F ) (Left) Relative Alk1 mRNA expression in colon tissues (RT-qPCR normalized to Gapdh). (Right) ALK1 protein concentrations (quantified by ELISA). Data presented as mean ± SD (* p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001 ns: not significant). ( G ) Relative mRNA expression levels of IL-1β, Ccl2, Col1a1, and Col3a1 (RT-qPCR; mean ± SD). ( H ) Western blot analysis of CCL2, TGF-β, and α-SMA protein expression in colon tissues and quantitative analysis of band intensities. ( I ) Western blot detection of p-Smad1, total Smad1, and VE-cadherin proteins and quantitative analysis of band intensities.

Article Snippet: Immediately after seeding, HIMECs were treated with the following stimuli: recombinant human BMP9 (rhBMP9; 0–10 ng/mL), TNF-α (20 ng/mL; Novoprotein, Suzhou, China, #C008), and ML347 (150 nM; MCE, Monmouth Junction, NJ, USA, #HY-12274).

Techniques: Recombinant, Control, Enzyme-linked Immunosorbent Assay, Staining, Activity Assay, Expressing, Quantitative RT-PCR, Western Blot

Journal: Biomedicines

Article Title: Recombinant BMP9 Reinforces Gut Vascular Barrier in Experimental Colitis

doi: 10.3390/biomedicines14020288

Figure Lengend Snippet: BMP9 restores intestinal vascular barrier integrity in DSS-induced colitis. ( A ) Representative immunofluorescence images of VE-cadherin (red) and CD31 (green) co-localization in colon tissues (nuclei counterstained with DAPI). ( B ) Left: Schematic of FITC-dextran (4 kDa) permeability assay. Right: Quantified serum FITC fluorescence intensity 60 min post-gavage ( n = 5 per group). ( C ) Left: Schematic of Evans Blue vascular leakage assay. Right: Colonic Evans Blue extravasation quantified by absorbance at 620 nm (mean ± SD; n = 5 per group; ** p < 0.01, *** p < 0.001, ns: not significant; one-way ANOVA with Tukey’s test). ( D ) KEGG pathway analysis of RNA-seq data from colon tissues (DSS + BMP9 groups vs. DSS). ( E ) Volcano plot of differentially expressed genes. Genes highlighted in bold are key IBD-associated downregulated factors.

Article Snippet: Immediately after seeding, HIMECs were treated with the following stimuli: recombinant human BMP9 (rhBMP9; 0–10 ng/mL), TNF-α (20 ng/mL; Novoprotein, Suzhou, China, #C008), and ML347 (150 nM; MCE, Monmouth Junction, NJ, USA, #HY-12274).

Techniques: Immunofluorescence, Permeability, Fluorescence, RNA Sequencing

Journal: Biomedicines

Article Title: Recombinant BMP9 Reinforces Gut Vascular Barrier in Experimental Colitis

doi: 10.3390/biomedicines14020288

Figure Lengend Snippet: BMP9/ALK1 signaling modulates neutrophil migration and endothelial tube formation. ( A ) Schematic representation of the neutrophil migration assay. ( B ) (Left) Representative fluorescence micrographs demonstrating Calcein-AM-labeled neutrophil migration through 3 μm pore Transwell inserts toward HIMECs pretreated for 2 h with: vehicle control (0 ng/mL BMP9), BMP9 (0.1, 1, or 10 ng/mL), TNF-α (20 ng/mL as positive control), or ALK1 inhibitor ML347 (150 nM). (Right) Quantitative analysis of neutrophil migration rates (mean ± SD; * p < 0.05, ** p < 0.01, *** p < 0.001, ns: not significant by two-way ANOVA with post hoc testing). ( C ) Schematic illustration of the endothelial tube formation assay protocol. ( D ) Representative phase-contrast images of tubular network formation by HIMECs cultured on growth factor-reduced Matrigel under various treatment conditions: vehicle control (0 ng/mL BMP9), BMP9 (0.1, 1, or 10 ng/mL), TNF-α (20 ng/mL), or ML347 (150 nM). ( E ) Quantitative assessment of angiogenic parameters including branch points and nodal junctions (mean ± SD; * p < 0.05, ** p < 0.01, *** p < 0.001 by two-way ANOVA with appropriate post hoc comparisons).

Article Snippet: Immediately after seeding, HIMECs were treated with the following stimuli: recombinant human BMP9 (rhBMP9; 0–10 ng/mL), TNF-α (20 ng/mL; Novoprotein, Suzhou, China, #C008), and ML347 (150 nM; MCE, Monmouth Junction, NJ, USA, #HY-12274).

Techniques: Migration, Fluorescence, Labeling, Control, Positive Control, Endothelial Tube Formation Assay, Cell Culture

Journal: JOR Spine

Article Title: In silico modeling the potential clinical effect of growth factor treatment on the metabolism of human nucleus pulposus cells

doi: 10.1002/jsp2.1352

Figure Lengend Snippet: (A) Human nucleus pulposus (NP) cells were isolated from four patients undergoing discectomy procedures. Following monolayer expansion, cell spheroids were formed in 96‐well plates coated with a thin layer of 2% agarose to prevent attachment. (B) Cell spheroids were cultured in standard expansion media (XPAN) for 1 week prior to 2 weeks under growth differentiation factor‐5 (GDF‐5) stimulation. (C) After GDF‐5 stimulation, individual spheroids underwent metabolic analysis using a Seahorse XFe96 analyzer and biochemical analysis. (D) Metabolic rates for each experimental group were then computed in silico to predict the donor‐specific reparative effects of GDF‐5 on extracellular matrix and subsequent impact on the nutrient microenvironment.

Article Snippet: Experimental groups consisted of a XPAN‐only (LG‐DMEM supplemented with 10% FBS and 2% Pen‐Strep) control (ctr) and the three clinically investigated GDF‐5 (PeproTech, Thermo Fisher Scientific) doses, consisting of XPAN supplemented with 0.25, 1, and 2 mg. Doses were normalized to the average cell population of the human NP to yield a working concentration of 0.75, 3, and 6 μg per spheroid for the 0.25, 1, and 2 mg groups, respectively.

Techniques: Isolation, Cell Culture, In Silico

Journal: JOR Spine

Article Title: In silico modeling the potential clinical effect of growth factor treatment on the metabolism of human nucleus pulposus cells

doi: 10.1002/jsp2.1352

Figure Lengend Snippet: (A) Spheroid viability was assessed across the experimental groups using live/dead staining to ensure viability remained high in each group prior to performing metabolic rate measurements. (B) Oxygen consumption rates (OCR, nmol/million cells/h) and (C) lactate production rates (LPR, nmol/million cells/h) for NP cell spheroids in either XPAN media as an untreated control (ctr) or stimulated with three different concentrations of growth differentiation factor‐5 (GDF‐5) (0.25, 1, and 2 mg) ( N = 4). While there is an apparent trend of increasing OCR and LPR with GDF‐5 concentration, only the 2 mg group had a significantly higher OCR compared to the ctr ( p = 0.045).

Article Snippet: Experimental groups consisted of a XPAN‐only (LG‐DMEM supplemented with 10% FBS and 2% Pen‐Strep) control (ctr) and the three clinically investigated GDF‐5 (PeproTech, Thermo Fisher Scientific) doses, consisting of XPAN supplemented with 0.25, 1, and 2 mg. Doses were normalized to the average cell population of the human NP to yield a working concentration of 0.75, 3, and 6 μg per spheroid for the 0.25, 1, and 2 mg groups, respectively.

Techniques: Staining, Control, Concentration Assay

Journal: JOR Spine

Article Title: In silico modeling the potential clinical effect of growth factor treatment on the metabolism of human nucleus pulposus cells

doi: 10.1002/jsp2.1352

Figure Lengend Snippet: (A) Glycosaminoglycan (GAG) production rates and (B) collagen production rates for cell spheroids in either XPAN media (control [ctr]) or stimulated with three different concentrations of growth differentiation factor‐5 (GDF‐5) (0.25, 1, and 2 mg) ( N = 4). While no statistically significant difference was detected between experimental groups, donor‐specific colors are used to highlight donor variability and trends in donor‐specific response across both GAG and collagen. (C) Corresponding histological evaluation for Donor 2 using hematoxylin and eosin (H&E) to stain for cells, alcian blue (AB) to stain for GAG and picrosirius red (PSR) to stain for collagen. Scale bar is 200 μm.

Article Snippet: Experimental groups consisted of a XPAN‐only (LG‐DMEM supplemented with 10% FBS and 2% Pen‐Strep) control (ctr) and the three clinically investigated GDF‐5 (PeproTech, Thermo Fisher Scientific) doses, consisting of XPAN supplemented with 0.25, 1, and 2 mg. Doses were normalized to the average cell population of the human NP to yield a working concentration of 0.75, 3, and 6 μg per spheroid for the 0.25, 1, and 2 mg groups, respectively.

Techniques: Control, Staining

Journal: iScience

Article Title: Time- and cell-specific activation of BMP signaling restrains chondrocyte hypertrophy

doi: 10.1016/j.isci.2024.110537

Figure Lengend Snippet:

Article Snippet: Recombinant GDF-5 , Peprotech , Cat. No. 120-01.

Techniques: Recombinant, Plasmid Preparation, RNA Sequencing Assay, Software