Journal: Journal of Advanced Research

Article Title: Novel function of macrophage migration inhibitory factor in regulating post-infarct inflammation and the therapeutic significance

doi: 10.1016/j.jare.2025.05.030

Figure Lengend Snippet: Macrophage migration inhibitory factor (MIF) promotes inflammatory cell migration and infiltration through C – C motif chemokine receptor 2 (CCR2) and C-X-C motif chemokine receptor (CXCR) 4 and promotes splenic monocyte mobilization via interaction with angiotensin Ⅱ type 1 receptor (AT-1R) after myocardial infarction (MI). A, peripheral blood mononuclear cells (PBMCs) chemotaxis in response to homogenized sham or infarct heart tissue from wild type (WT) mice at day 3 post-MI, or MI homogenate together with inhibition of C–C motif chemokine receptor 2 (CCR2), C-X-C motif chemokine receptor (CXCR) 2 and CXCR4, respectively. Cells/HPF, cells/high power field. PBMCs harvested from WT mice at 3 days post-MI. CCR2i, CCR2 inhibition. CXCR2i, CXCR2 inhibition. CXCR4i, CXCR4 inhibition. n = 4–8 per group. *** P < 0.001 vs. sham . B, PBMCs chemotaxis in response to recombinant human MIF (rMIF) or rMIF with inhibition of CCR2, CXCR2 and CXCR4, respectively. PBMCs harvested from WT mice at 3 days post-MI. n = 4–8 per group. *** P < 0.001 vs. control . C, Representative immunoblotting images for monocyte chemoattractant protein-1 (MCP-1), CXCR4 and internal reference protein (heat shock protein 60, HSP-60) in hearts from WT and global MIF deficient (MIFKO) mice with sham-operation or 72 h MI. D-E, Quantitative analysis of MCP-1 and CXCR4 expression. n = 6–9 per group. ** P < 0.01 vs. sham in the same genotype. *** P < 0.001 vs. sham in the same genotype. F, Splenic monocyte (from normal WT mice) chemotaxis in response to plasma from normal mice or mice with 1.5 h MI, or MI plasma together with losartan. Lor, losartan, AT-1R inhibitor. n = 6–8 per group. *** P < 0.001 vs. normal. G, Splenic monocytes (from normal WT mice) chemotaxis in response to rMIF and angiotensin II (Ang II), or together with losartan addition, respectively. n = 6–8 per group. ** P < 0.01 vs. Control. *** P < 0.001 vs. Control. H, Representative immunoblotting images for AT-1R and internal reference protein (HSP-60) in spleens from WT and global MIF deficient (MIFKO) mice with sham-operation and 1.5 h MI. I, Quantitative analysis of AT-1R expression. n = 8 per group. *** P < 0.001 vs. sham in the same genotype . J. Co-immunoprecipitation (Co-IP) assay using an anti-AT-1R antibody or IgG demonstrated the interaction between AT-1R and MIF, as well as between AT-1R and CD74 in splenic monocytes and spleen tissue. Input, input sample. IgG, negative control sample. IP, Co-IP sample.

Article Snippet: Primary antibodies against MCP-1 (1:1000), CCR2 (1:1000), CXCR2 (1:1000), CXCR4 (1:1000), tissue inhibitor of metalloproteinases-4 (Timp-4, 1:1000), AT-1R (AGTR-1, 1:1000), heat shock protein 60 (HSP-60, 1:1000) and HSP-90 (1:1000) were purchased from ProteinTech.

Techniques: Migration, Chemotaxis Assay, Inhibition, Recombinant, Control, Western Blot, Expressing, Clinical Proteomics, Co-Immunoprecipitation Assay, Negative Control

Journal: PLoS ONE

Article Title: Cell Population Kinetics of Collagen Scaffolds in Ex Vivo Oral Wound Repair

doi: 10.1371/journal.pone.0112680

Figure Lengend Snippet: The ex vivo wound healing model is shown in top view and cross-sectional view (A). The bottom of the well was covered with an acellular growth factor reduced basal membrane extract (BME). The scaffold (sponge type scaffold composed of cross-linked collagen or the gel type scaffold composed of rat collagen I) in the center was surrounded by a BME-gel containing 800,000 hGFs/ml and covered with an acellular BME gel and Dulbecco's Modified Eagle's Medium (DMEM) supplemented with 1% fetal bovine serum (FBS) and antibiotics. Both scaffold and gel were tested with and without the addition of platelet-derived growth factor-BB (PDGF). Fluorescence images were taken to assess DiI-labeled cells within the scaffold over time. To assess the metabolic activity and gene expression dynamics, the scaffolds including the “active zone” (dashed line) were subjected to MTT tests, gene array analysis, and quantitative PCR. (B) Representative scanning electron microscopy images depict the morphology of the collagen gel, collagen scaffold at 400×, and 800× magnification. The white bar represents 100 µm.

Article Snippet: 48 well plates were first layered with 50 μL of 12 mg/ml growth factor reduced basal membrane extract (BME; CULTREX, TREVIGEN, Gaithersburg, MD, USA) without cells.

Techniques: Ex Vivo, Membrane, Modification, Derivative Assay, Fluorescence, Labeling, Activity Assay, Gene Expression, Real-time Polymerase Chain Reaction, Electron Microscopy