sc-43  (MedChemExpress)

Journal: Journal of Sport and Health Science

Article Title: Exercise training-induced extracellular miR-136-3p modulates glucose uptake and myogenesis through targeting of NRDC in human skeletal muscle

doi: 10.1016/j.jshs.2025.101091

Figure Lengend Snippet: NRDC is a direct target of miR-136-3p in human myotubes. Skeletal muscle NRDC mRNA is responsive to training and inactivity. (A) Tissue mRNA expression of NRDC from the Human Protein Atlas database showing enriched expression of NRDC in human skeletal muscle. (B) The miR-136-3p target site in the NRDC gene is highly conserved in mammals. (C) Luciferase activity in HEK293 cells co-transfected the NRDC 3’UTR and miR-136-3p with or without anti-miR136-3p inhibitors. miR-136-3p transfection downregulates NRDC (D) mRNA and (E) representative image of protein abundance in human myotubes. (F) Publicly available data ( GSE14413 ) showing NRDC mRNA expression in human skeletal muscle of healthy young participants after 6 weeks of endurance training ( n = 8). (G) Publicly available data ( GSE120862 ) showing NRDC mRNA expression in human skeletal muscle of healthy young participants after 2 months of aerobic training ( n = 10). (H) Publicly available data ( GSE14901 ) showing NRDC mRNA expression in human skeletal muscle of healthy young participants after 14 days of immobilization ( n = 24). * p < 0.05, ** p < 0.005. GSE = gene set enrichment; HEK293 = human embryonic kidney; miR = microRNA; NC = negative control; NRDC = nardilysin convertase; nTPM = normalized transcripts per million; si NRDC = small interfering RNA of NRDC ; UTR = untranslated region.

Article Snippet: Membranes were incubated with primary antibodies directed to NRDC (sc-137199; Santa Cruz Biotechnology, Dallas, TX, USA), AMP-activated protein kinase α (AMPKα) (#2532; Cell Signaling Technology, Danvers, MA, USA), Akt substrate of 160 kDa (AS160) (#2670; Cell Signaling Technology), phospho-AS160 (#8730; Cell Signaling Technology), protein kinase B (AKT) (#9272; Cell Signaling Technology), phospho-AKT (Ser473) (#9271; Cell Signaling Technology), or puromycin (MABE343; Merck Burlington, MA, USA).

Techniques: Expressing, Luciferase, Activity Assay, Transfection, Quantitative Proteomics, Negative Control, Small Interfering RNA

Journal: Bioactive Materials

Article Title: Tissue-specific matrix-bound nanovesicles regulate the immunoregulatory progress of biological mesh-aided abdominal hernia repair

doi: 10.1016/j.bioactmat.2026.03.004

Figure Lengend Snippet: Structural and biological characterization of SIS and UBM-SIS meshes and isolated MBVs. A) Surface and cross-sectional morphology of SIS and UBM-SIS meshes by SEM. B) Quantification of mesh thickness (n = 10). C) Pore size and porosity analysis of mesh (n = 10). D) Live/dead staining of fibroblasts on SIS and UBM-SIS at day 1, 4, and 7. (E) CCK-8 assay of fibroblast viability on meshes (n = 5). F) Immunofluorescence staining of fibroblasts (TGF-β1, day 3), SMCs (phalloidin, day 7) and HUVECs (CD31, day 14) on SIS and UBM-SIS meshes, and SEM and DAPI staining of SMCs (day 21) coverage and cellular infiltration. White dashed lines delineate the upper and lower boundaries of the ECM scaffold. The yellow dashed line indicates the infiltration depth, defined as the distance from the scaffold surface to the DAPI-positive nucleus formed as a cellular floor and used for quantitative analysis. G) Quantification of TGF-β1, cytoskeletal area (phalloidin) and CD31 expression (n = 5). H) Quantification of cellular infiltration across mesh thickness (n = 5). I) Schematic of ECM components retained in decellularized ECM mesh. J) H&E and Masson's trichrome staining of mesh. K) Residual DNA quantification. L) Analysis of cytokine and growth factor profiling upon mesh-specific difference (n = 4). M) Workflow for MBV isolation and analysis. N) SEM images of MBV embedded on ECM. O) TEM images of MBV morphology. P) NTA analysis of MBV (n = 4). Q) Western blot detection of exosomal markers in MBV. Data are presented as mean ± SD. Each dot represents an independent sample. Statistical significance was determined using two-tailed unpaired Student's t-test for comparisons between two groups (B, C, and K), or one-way ANOVA followed by Tukey's post hoc test for multiple comparisons (E, G, and H), where ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001 and ∗∗∗∗ p < 0.0001.

Article Snippet: Additional antibodies, such as mouse polyclonal anti -TGF-β1, anti-elastin, CD11b, CD68, CD86, and CD206, along with the BCA Protein Assay Kit, were obtained from Santa Cruz Biotechnology (Dallas, TX, USA).

Techniques: Isolation, Pore Size, Staining, CCK-8 Assay, Immunofluorescence, Expressing, Western Blot, Two Tailed Test

Journal: Bioactive Materials

Article Title: Tissue-specific matrix-bound nanovesicles regulate the immunoregulatory progress of biological mesh-aided abdominal hernia repair

doi: 10.1016/j.bioactmat.2026.03.004

Figure Lengend Snippet: Bioactivity and immunomodulatory properties of MBVs derived from SIS and UBM-SIS meshes. A) Schematic illustration of MBV-regulated cellular activities during ECM remodeling via their interactions with fibroblasts, SMCs, HUVECs, and macrophages to validate MBVs as bioactive components embedded within parent ECM. Nuclei are labeled with DAPI (blue); PKH26 (red) marks MBVs; phalloidin (green) stains F-actin. B) Immunofluorescence staining of fibroblasts (TGF-β1, collagen I), SMCs (phalloidin), and HUVECs (CD31) after treatment with SIS MBVs or UBM-SIS MBVs. C) Quantification of fluorescence signal area per cell for respective markers (n = 5). D) Western blot analysis of marker proteins in MBV-treated cells. E) Relative protein expression levels normalized to GAPDH (n = 3). F) Schematic of macrophage polarization model with/without MBV treatment. G) Immunostaining of macrophages for F4/80, iNOS (M1-like), and Arg-1 (M2-like) under different stimulations and MBV-treated conditions. H) Quantification of mean fluorescence intensity (MFI) of iNOS and Arg-1 (n = 5). I) Western blot analysis of pro- and anti-inflammatory markers in MBV-treated macrophages and LPS + IFN-γ-treated macrophages (control). J) Quantification of relative protein levels (n = 3). K) Heatmap of RT-qPCR analysis showing cytokine and ECM regulator gene expression in MBV-treated macrophages (n = 3). L) Representative fluorescence images of DCFH staining in macrophages following different treatments. M) Quantification of DCFH fluorescence area per cell (n = 5). Data are presented as mean ± SD. Each dot represents an independent biological replicate. Statistical significance was assessed using one-way ANOVA followed by Tukey's post hoc test, where ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001 and ∗∗∗∗ p < 0.0001.

Article Snippet: Additional antibodies, such as mouse polyclonal anti -TGF-β1, anti-elastin, CD11b, CD68, CD86, and CD206, along with the BCA Protein Assay Kit, were obtained from Santa Cruz Biotechnology (Dallas, TX, USA).

Techniques: Derivative Assay, Labeling, Immunofluorescence, Staining, Fluorescence, Western Blot, Marker, Expressing, Immunostaining, Control, Quantitative RT-PCR, Gene Expression

Journal: Bioactive Materials

Article Title: Tissue-specific matrix-bound nanovesicles regulate the immunoregulatory progress of biological mesh-aided abdominal hernia repair

doi: 10.1016/j.bioactmat.2026.03.004

Figure Lengend Snippet: Functional comparison of MBVs derived from SIS and UBM in modulating inflammation, angiogenesis, and matrix remodeling. A) Schematic of macrophage polarization model under LPS + IFN-γ stimulation with or without MBV treatment. B) Immunofluorescence staining of iNOS (red) and Arg-1 (green) in macrophages treated with SIS MBVs, UBM MBVs, or UBM-SIS MBVs. C) Quantification of mean fluorescence intensity (MFI) of iNOS and Arg-1 (n = 5). D–G) RT-qPCR analysis of pro- (( TNF-α, IL-6 ) and anti-inflammatory ( IL-10, TGF-β1 ) cytokine gene expression in MBV-treated macrophages (n = 4). H) Schematic of analysis of MBV-treated HUVECs and fibroblasts cultured in Matrigel. I) 3D immunostaining of CD31 + tube-like structures in HUVECs after MBV treatment. J–K) Quantification of tube-like area percentage and number of tube-like structures per field (n = 6). L) 3D two-photo images of TGF-β1 expression in fibroblasts cultured with different MBVs. M) Quantification of TGF-β1-positive volume percentage in fibroblasts (n = 4). N-P) Western blot analysis of NF-κB and STAT3 pathway proteins in MBV-treated macrophages, angiogenic signaling proteins (AKT, ERK1/2) in MBV-treated HUVECs, TGF-β/Smad signaling pathway in MBV-treated fibroblasts. Quantification of respective signaling molecules (n = 3). Data are presented as mean ± SD. Each dot represents an independent biological replicate. Statistical significance was assessed using one-way ANOVA followed by Tukey's post hoc test for multiple comparisons, where ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, and ∗∗∗∗ p < 0.0001.

Article Snippet: Additional antibodies, such as mouse polyclonal anti -TGF-β1, anti-elastin, CD11b, CD68, CD86, and CD206, along with the BCA Protein Assay Kit, were obtained from Santa Cruz Biotechnology (Dallas, TX, USA).

Techniques: Functional Assay, Comparison, Derivative Assay, Immunofluorescence, Staining, Fluorescence, Quantitative RT-PCR, Gene Expression, Cell Culture, Immunostaining, Expressing, Western Blot

Journal: Bioactive Materials

Article Title: Tissue-specific matrix-bound nanovesicles regulate the immunoregulatory progress of biological mesh-aided abdominal hernia repair

doi: 10.1016/j.bioactmat.2026.03.004

Figure Lengend Snippet: Inflammatory immune responses following mesh implantation. A) Schematic illustration and representative macroscopic images of seroma tissues collected from the explants at 1 week. B) ELISA analysis of cytokines in the seroma fluid after 1 week (n = 4). C) Representative immunofluorescence images of CD11b + cell infiltration in mesh at 1 and 4 weeks. Scale bars: left, 1000 μm; right, 50 μm. D–E) Quantification of CD11b + cell density (n = 5, 4 samples per rat). F) Representative immunofluorescence images of CD68 (yellow), CD86 (green), and CD206 (red) staining of SIS and UBM-SIS at 1 and 4 weeks. Scale bars: top, 1000 μm; bottom, 50 μm. G–H) Quantification of CD68 + macrophage infiltration and M2-like/M1-like phenotypic distribution at 1 week (n = 5). I) Statistical comparison of M2-like/M1-like ratios between SIS and UBM-SIS groups (n = 5). J–K) CD68 + macrophage infiltration and CD206 + /CD86 + phenotypic distribution at 4 weeks (n = 5, 4 samples per rat). L) Quantification of M2-like/M1-like ratios at 4 weeks (n = 5, 4 samples per rat). M−O) Representative immunofluorescence images of iNOS and Arg-1 at tissue-mesh interfaces at 1 and 4 weeks, with quantitative analysis of positive area (n = 5, with 4 samples per rat). Scale bars: 50 μm. P) RT-qPCR analysis of pro- (( TNF-α, IL-6 ) and anti-inflammatory ( IL-10, TGF-β1 ) cytokine gene expression in SIS and UBM-SIS explants. Q) Schematic summary of immune response transition induced by SIS versus UBM-SIS MBV-containing meshes over 4 weeks. The asterisk indicates the implanted mesh. Data are presented as mean ± SD. For (B, I, L, and P), mean value of each rat (n = 5) was used for statistical comparisons. For (D, E, G, H, J, K, N, and O), each dot represents one section-level sample, where the value of each animal for statistical comparisons was obtained by averaging measurements from 4 samples. Statistical comparisons were performed within each time point using two-tailed unpaired Student's t-test, where ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001.

Article Snippet: Additional antibodies, such as mouse polyclonal anti -TGF-β1, anti-elastin, CD11b, CD68, CD86, and CD206, along with the BCA Protein Assay Kit, were obtained from Santa Cruz Biotechnology (Dallas, TX, USA).

Techniques: Enzyme-linked Immunosorbent Assay, Immunofluorescence, Staining, Comparison, Quantitative RT-PCR, Gene Expression, Two Tailed Test

Journal: Bioactive Materials

Article Title: Tissue-specific matrix-bound nanovesicles regulate the immunoregulatory progress of biological mesh-aided abdominal hernia repair

doi: 10.1016/j.bioactmat.2026.03.004

Figure Lengend Snippet: ECM remodeling and mechanical evaluation of meshes and explants. A) Representative immunofluorescence images showing collagen I (red) and collagen III (green) deposition in the center and interface regions of SIS and UBM–SIS explants at 8 weeks. Scale bars: overview = 1000 μm, magnified images = 100 μm. B–E) Quantification of total collagen (I + III), collagen I, and collagen III positive expression and collagen I/III ratio in the center and interface regions (n = 5, 4 samples per rat). F–G) Polar plot of collagen fiber orientation in the center and interface regions analyzed by orientation distribution. H) Orientation coherency of collagen fibers in the center and interface regions (n = 5, 4 samples per rat). I) Aspect ratio analysis indicating collagen fibril anisotropy (n = 4). J–K) Representative immunofluorescence staining of TGF-β1 and α-SMA at 8 weeks and corresponding quantification of positive area (n = 5, 4 samples per rat). L, M) Mechanical characterization of meshes and explants showing ultimate tensile strength and elongation at break across different time points (n = 4). N) Comparison of tensile strength and elongation of explants with native abdominal wall components, including posterior and anterior rectus sheath, linea alba, peritoneum, and transversalis fascia. The asterisk indicates the implanted mesh. Data are presented as mean ± SD. For (B-E, H, and K), mean value of each rat (n = 5) was used for statistical comparisons. Each dot represents one section-level sample, where the value for each animal was obtained by averaging measurements from 4 samples. For (I, L, and M), mean value of each rat (n = 4) was used for statistical comparisons. Statistical significance was determined using two-tailed unpaired Student's t-test, where ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001.

Article Snippet: Additional antibodies, such as mouse polyclonal anti -TGF-β1, anti-elastin, CD11b, CD68, CD86, and CD206, along with the BCA Protein Assay Kit, were obtained from Santa Cruz Biotechnology (Dallas, TX, USA).

Techniques: Immunofluorescence, Expressing, Staining, Comparison, Two Tailed Test

Journal: Bioactive Materials

Article Title: Tissue-specific matrix-bound nanovesicles regulate the immunoregulatory progress of biological mesh-aided abdominal hernia repair

doi: 10.1016/j.bioactmat.2026.03.004

Figure Lengend Snippet: Transcriptomic analysis of explants from different groups after 1 week and comparative analysis of MBV and ECM properties. A) Schematic illustration of tissues collected from the explants at 1 week for distinct signaling pathway analysis. B) Volcano plot showing differentially expressed genes (DEGs) between UBM-SIS and SIS groups (n = 3 per group). C) Heatmap of DEGs between SIS and UBM-SIS groups (red: upregulated, blue: downregulated; cutoff >1.0; n = 3). D) KEGG pathway enrichment analysis of downregulated genes in UBM-SIS compared to SIS. E) Reactome pathway enrichment analysis of downregulated genes in UBM-SIS compared to SIS. F–H) GSEA demonstrating altered gene signatures related to NET formation, NF-κB pathway and cytokine-cytokine receptor interaction. I) Radar plot comparing SIS- and UBM-derived MBVs. The five axes represent key pathways involved in angiogenesis ( ERK ), vascularization ( AKT ), inflammation ( p65 ), immunomodulation ( STAT3 ), and remodeling ( TGF-β/Smad ). J) Radar plot summarizing ECM in vivo performance at 1 and 4 weeks. The five axes represent essential features in ECM remodeling, including adhesion, angiogenesis, inflammation, immunomodulation, and collagen deposition.

Article Snippet: Additional antibodies, such as mouse polyclonal anti -TGF-β1, anti-elastin, CD11b, CD68, CD86, and CD206, along with the BCA Protein Assay Kit, were obtained from Santa Cruz Biotechnology (Dallas, TX, USA).

Techniques: Derivative Assay, In Vivo