Journal: TH Open: Companion Journal to Thrombosis and Haemostasis

Article Title: Effects of DOACs on Mouse Melanoma Metastasis and the Inhibitory Mechanism of Edoxaban, a Factor Xa-Specific DOAC

doi: 10.1055/a-2701-4242

Figure Lengend Snippet: Possible mechanism of melanoma metastasis mediated by FXa-PAR2 and FXa-PAR2-TGFβ pathways in mice implanted with B16 melanoma cells. ( A ) In the absence of EDX, FXa produced by activation of the blood coagulation system and/or cancer-associated FXa-like protease in hosts implanted with melanoma cells activates the FXa-PAR2 and FXa-PAR2-TGFβ pathways in living tissue and melanoma cells, resulting in a decrease in tight junction-associated factors (claudin 5 and E-cadherin) between tissue cells, and an increase in other tumor-associated factors, thereby promoting inflammation, tumor angiogenesis, tissue invasion, and EMT in the host body, and promoting metastasis of melanoma cells. ( B ) In mice treated with EDX, activation of the FXa-PAR2 and FXa-PAR2-TGFβ pathways is suppressed, tight junction-associated factors are increased, and other angiogenic, invasion, and EMT-associated factors are decreased, resulting in the suppression of inflammation, angiogenesis, tissue invasion, and EMT, and the inhibition of melanoma cell metastasis. Ang-2, angiopoietin-2; bFGF, basic fibroblast growth factor; EDX, edoxaban; EMT, epithelial–mesenchymal transition; FXa, factor Xa; IL-6, interleukin 6; MMP, matrix metalloproteinase; PAR, protease-activated receptor; PDGF, platelet-derived growth factor; SMAD, small mothers against decapentaplegic; Snail-1, small family zinc finger 1; TGFβ1, transforming growth factor β1; TGFβR I, TGFβ receptor type I; TGFβR II, TGFβR type II; VEGFA, vascular endothelial growth factor A; Wnt3a, wingless MMTV integration site family, member 3a; ZEB1, zinc finger E-box binding homeobox 1.

Article Snippet: The levels of interleukin 6 (IL-6), protease-activated receptor (PAR) 1 (PAR1), PAR2, transforming growth factor β1 (TGFβ1), TGFβ receptor type I (TGFβR I), small mothers against decapentaplegic (SMAD) family member 2 (SMAD2), SMAD3, SMAD4, matrix metalloproteinase (MMP)-2 (MMP-2), MMP-9, angiopoietin-2, basic fibroblast growth factor (bFGF), vimentin, fibronectin, snail family transcriptional repressor 1 (Snail-1), inducible nitric oxide synthase (iNOS), and arginase-1 in the lung were determined using commercially available enzyme-linked immunosorbent assay kits per manufacturers' instructions: IL-6 (M6000B, R&D Systems) and angiopoietin-2 (MANG20, R&D Systems), PAR1 (MBS753326, MyBioSoutce, San Diego, CA), PAR2 (MBS4501658; MyBioSource), TGFβ1 (E-EL-M0051, Elabscience, Houston, TX), TGFβR I ( Q64729 , RayBiotech Life, Peachtree Corners, GA), SMAD2 (OKEH03472, Aviva Systems Biology, San Diego, CA), SMAD3 (OKEH03473, Aviva Systems Biology), SMAD4 (OKEH03425, Aviva Systems Biology), fibronectin (OKCD05702, Aviva Systems Biology), MMP-2 (ab254516, Abcam), MMP-9 (ab253227, Abcam), bFGF (bs-0217R, Bioss Antibodies), vimentin (ELK3731, ELK Biotechnology, Denver, CO), Snail-1 (LS-F2317-1, LS Bio, Shirley, MA), iNOS (CSB-E08326m, Wuhan Fine Biotech, Hubei, China), and arginase-1 (MBS2882661, MyBioSource).

Techniques: Produced, Activation Assay, Coagulation, Inhibition, Derivative Assay, Binding Assay

Journal: Bioactive Materials

Article Title: Bioinspired scaffold recapitulating chondrogenic ontogeny and microenvironment for functional cartilage regeneration

doi: 10.1016/j.bioactmat.2025.11.041

Figure Lengend Snippet: Spatiotemporal remodeling of FGF signaling networks in chondrocyte-like subpopulations. A) Heatmap of FGF signaling mediated communication patterns across chondrocyte-like subpopulations. B) Circle plot visualizing differential inter-subpopulation communication of FGF ligand-receptor signaling in chondrocyte-like cells at 4 week. C) Circle plot visualizing differential inter-subpopulation communication of FGF ligand-receptor signaling in chondrocyte-like cells at 8 week. D) Violin plots of expressions of FGF genes. E) Bubble plot of dysregulated ligand-receptor pairs. F) The differences in the overall information flow in the network. G) Representative western blots showing protein levels of FGF2 and FGF18 in Control and CE-SKP co-cultured cells.

Article Snippet: After blocking the membranes for an hour at room temperature using fast blocking solution, the membranes were incubated with primary antibodies specific for SOX9 (1:2000, 67439-1-Ig, Proteintech), COL2 (1:200, NBP1-91056, Novus Biologicals), COL10 (1:500, 26984-1-AP, Proteintech), COL1 (1:2000, 67288-1-Ig, Proteintech), RUNX2 (1:200, 20700-1-AP, Proteintech), FGF2 (1:500, 11234-1-AP, Proteintech), FGF18 (1:500, 60341-1-Ig, Proteintech) and GAPDH (1:50000, 60004-1-Ig, Proteintech) for an entire night at 4 °C.

Techniques: Western Blot, Control, Cell Culture

Journal: Biomaterials Research

Article Title: Derivation of Mesenchymal Stem Cells through Sequential Presentation of Growth Factors via Gelatin Microparticles in Pluripotent Stem Cell Spheroids

doi: 10.34133/bmr.0184

Figure Lengend Snippet: Bone morphogenetic protein-4 (BMP4)- and fibroblast growth factor 2 (FGF2)-induced differentiation of induced pluripotent stem cells (iPSCs) to mesenchymal stem cells (MSCs) in 2-dimensional (2D) conditions. (A) Schematic diagram depicting iPSC-to-induced MSC (iMSC) differentiation via mesoderm induction. (B) Images of iPSCs, mesoderm-induced cells, and iMSCs. Scale bar = 200 μm. (C) Quantitative real-time gene analysis (qPCR; top panel) and western blot (WB) analysis (bottom panel) of the mesoderm markers Brachyury and SNAI1 (Snail1), along with the pluripotency marker SOX2, at 4 d postdifferentiation. (D) qPCR (top panel) and WB analysis (bottom panel) of the MSC markers CD73, CD90, CD105, and CD44 at 11 d postdifferentiation. (E) Flow cytometry (fluorescence-activated cell sorting [FACS]) analysis of the MSC markers CD73, CD90, and CD44 in cells treated with either BMP4 only or BMP4 + FGF2. The qPCR data were normalized to 18S. The WB data were normalized to β-actin. All data represent results from 3 independent experiments, each conducted in triplicate. The data are presented as mean ± SD (ns, not significant; * P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.0001). Individual data points and significance levels are indicated in graphs. DMEM, Dulbecco’s modified Eagle medium; SR, serum replacement; hiPSCs, human iPSCs; mRNA, messenger RNA; GF, growth factor.

Article Snippet: The slow- and fast-degrading GelMPs were incubated with primary antibodies against FGF2 (Santa Cruz Biotechnology, USA) and BMP4 (PeproTech, USA) overnight and labeled with Alexa Fluor 488 (Invitrogen, Thermo Fisher Scientific, USA) and Texas Red (Invitrogen, Thermo Fisher Scientific, USA) at 1 μl/100 μl of PBS, respectively.

Techniques: Western Blot, Marker, Flow Cytometry, Fluorescence, FACS, Modification

Journal: Biomaterials Research

Article Title: Derivation of Mesenchymal Stem Cells through Sequential Presentation of Growth Factors via Gelatin Microparticles in Pluripotent Stem Cell Spheroids

doi: 10.34133/bmr.0184

Figure Lengend Snippet: Gelatin microparticle (GelMP) fabrication, characterization, GF conjugation, and the release profiles of BMP4 and FGF2. (A) Scanning electron microscopy (SEM) images of fast-degrading (4 mM glutaraldehyde) and slow-degrading (12 mM glutaraldehyde) GelMPs. Scale bar = 100 μm. Size distributions of fast- and slow-degrading GelMPs. (B) Physical properties of fast- and slow-degrading GelMPs. (C) Representative immunofluorescent images of BMP4-conjugated fast-degrading GelMPs and FGF2-conjugated slow-degrading GelMPs. Scale bar = 100 μm. (D) Degradation rate of 5 mg of microparticles in collagenase (5 μg/ml) solution. (E) Cumulative release (%) of 1 μg of BMP4 from fast-degrading GelMPs and 1 μg of FGF2 from slow-degrading GelMPs incubated for 20 d in collagenase (5 μg/ml) solution. The obtained data represent results from 3 independent experiments, each conducted in triplicate. PBS, phosphate-buffered saline.

Article Snippet: The slow- and fast-degrading GelMPs were incubated with primary antibodies against FGF2 (Santa Cruz Biotechnology, USA) and BMP4 (PeproTech, USA) overnight and labeled with Alexa Fluor 488 (Invitrogen, Thermo Fisher Scientific, USA) and Texas Red (Invitrogen, Thermo Fisher Scientific, USA) at 1 μl/100 μl of PBS, respectively.

Techniques: Conjugation Assay, Electron Microscopy, Incubation, Saline