Journal: Bioactive Materials

Article Title: Geometry-driven immunomodulation in 3D-printed bioceramics: Negative curvature promotes macrophage M2 polarization via Ras-MAPK/HIF-1α signaling for vascularized osteogenesis

doi: 10.1016/j.bioactmat.2026.01.001

Figure Lengend Snippet: Macrophage polarization analysis of Raw264.7 on structures with different gaussian curvature: (A, B) Chord Diagram for qPCR analysis of CCR7, IL6, iNOS-inflammatory and M1 marker genes, and Arg-1, CD206, IL10-M2 related protein genes in different Gaussian curvature groups. (C) Protein content of Arg-1 in different Gaussian curvature groups at 1 and 3 days. (D) Integral plots of the five experimental groups. IL4 group is the positive control for CD206 expression and lipopolysaccharide (LPS) group is the negative control.

Article Snippet: The reagents used in the experiment included: H-DMEM(11965118, Gibco, USA.), α-DMEM medium(12571063, Gibco, USA.), TritonX-100(ST1723, Beyotime, China), 4 % paraformaldehyde (BL539A, Biosharp, China),FBS(A5256701, Gibco, USA.),ECM medium (Science Cell, USA.),and DAPI staining solution (C1006, Beyotime, China),BCIP/NBT(C3206, Beyotime, China), reactive oxygen species kit (S0033S, Beyotime, China), BSA (B2064, ≥98 %, Sigma-Aldrich, USA.),CD31 antibody (ab28364, Abcam, USA.), secondary anti-IGg (ab175773, Alexa Fluor® 680, Abcam, USA.), Phalloidin-iFluor 488(ab176753, Abcam, USA.), CCR7(AF5293, Bioss, China), CD206 (bsm-60761R, Bioss, China), iNOS (bs-22924R, Bioss, China), RIPA (P0013, Beyotime, China), p-ERK1/2 (AF3687, Affinity, USA.) and ERK1/2 (#AF0155, Affinity, USA), luminol detection reagent (sc-2048, Santa Cruz, USA.), GAPDH (Cat#KC-5G5, Kangchen Biotechnology, China), Trizol(15596026CN, Invitrogen, USA.), DEPC(R0601, Thermo Scientific, USA.), TBST(R017R.0000, Thermo Scientific, USA.), HIF-1a(GTX127309, GeneTex, USA.), β-Tubulin(10094-1-AP, Proteintech, UK) Adezmapimod (SB 203580, MCE, USA.) medium and Paclitaxel (99.88 %, HY-B0015R, MCE), ELISA Arg-1(E-EL-M3092, ELabSci@, China), TNF-α (E-EL-M3063, ELabSci@, China), OPN(22952-1-AP, Proteintech, UK.), F4/80 (GB11027-100, Servicebio, China) Alkaline phosphatase activity kit (P0321S, Beyotime, China), Matrigel (CLS356234, Corning, USA), Microfill MV120 (Flow tech, USA), EDTA(17892, Thermo Scientific, USA.) xylene(X112051, AR,99 %, Aladdin, China), ethanol (107-21-1, AR,99 %, Aladdin, China).

Techniques: Marker, Positive Control, Expressing, Negative Control

Journal: Bioactive Materials

Article Title: Immunomodulatory effects of biodegradable Mg–Cu–Zn alloy in esophageal cancer

doi: 10.1016/j.bioactmat.2026.02.046

Figure Lengend Snippet: Distribution of CD163 + M2 TAMs in AKR-derived allograft tumor tissues from immunocompetent C57BL/6 mice. (a, b) Representative IHC staining images showing CD163 + M2 TAMs in the (a) peritumoral stroma and (b) tumor islets. Lower panels display higher-magnification views of the regions outlined by red dashed boxes. (c, d) Quantification of CD163 + cells in the (c) peritumoral stroma and (d) tumor islets. (e) Comparison of CD163 + cell density between the peritumoral stroma and tumor islets. (f) Total number of CD163 + cells in allograft tumors (peritumoral stroma and tumor islets combined). (g, h) Comparison of the density between iNOS + cells and CD163 + cells in the (g) peritumoral stroma and (h) tumor islets. (i, j) Quantification of iNOS + /CD163 + ratio in the (i) peritumoral stroma and (j) tumor islets. p < 0.05 (∗), p < 0.01 (∗∗), p < 0.001 (∗∗∗). A field of view is ∼0.086 mm 2 in (c−j).

Article Snippet: Tissue sections were then incubated with primary antibodies against iNOS (22226-1-AP, ProteinTech, China), CD163 (A26411PM, Abclone, China), CD8 (SP16, Maixin, China), CD4 (SP35, Maixin, China) or Ki-67 (12202S, Cell Signaling Technology) for 12 h at 4 °C, followed by secondary antibodies (Beyotime Biotechnology, Nantong, China).

Techniques: Derivative Assay, Immunohistochemistry, Comparison

Journal: Bioactive Materials

Article Title: Immunomodulatory effects of biodegradable Mg–Cu–Zn alloy in esophageal cancer

doi: 10.1016/j.bioactmat.2026.02.046

Figure Lengend Snippet: Distribution of CD163 + M2 TAMs in AKR-derived allograft tumor tissues from immunocompetent C57BL/6 mice. (a, b) Representative IHC staining images showing CD163 + M2 TAMs in the (a) peritumoral stroma and (b) tumor islets. Lower panels display higher-magnification views of the regions outlined by red dashed boxes. (c, d) Quantification of CD163 + cells in the (c) peritumoral stroma and (d) tumor islets. (e) Comparison of CD163 + cell density between the peritumoral stroma and tumor islets. (f) Total number of CD163 + cells in allograft tumors (peritumoral stroma and tumor islets combined). (g, h) Comparison of the density between iNOS + cells and CD163 + cells in the (g) peritumoral stroma and (h) tumor islets. (i, j) Quantification of iNOS + /CD163 + ratio in the (i) peritumoral stroma and (j) tumor islets. p < 0.05 (∗), p < 0.01 (∗∗), p < 0.001 (∗∗∗). A field of view is ∼0.086 mm 2 in (c−j).

Article Snippet: Tissue sections were then incubated with primary antibodies against iNOS (22226-1-AP, ProteinTech, China), CD163 (A26411PM, Abclone, China), CD8 (SP16, Maixin, China), CD4 (SP35, Maixin, China) or Ki-67 (12202S, Cell Signaling Technology) for 12 h at 4 °C, followed by secondary antibodies (Beyotime Biotechnology, Nantong, China).

Techniques: Derivative Assay, Immunohistochemistry, Comparison

Journal: Bioactive Materials

Article Title: Skin-mimetic bilayer hydrogel normalizes diabetic wound healing by orchestrating inflammatory cell dynamics: An early intervention strategy

doi: 10.1016/j.bioactmat.2026.02.025

Figure Lengend Snippet: In vitro assay of inflammation cell modulation under stimulation of SP-loaded Gel/HA and IL-10-loaded Ker/Cu. a Schematic of neutrophil migration test using a transwell system after treatment with the leaching solution of SP@Gel/HA. Gel/HA and blank culture medium were set for comparison. b Wright-Giemsa staining of HL-60 cells before and after differentiation. c Photograph of dHL-60 cells migrating to the lower chamber. d Quantitative analysis of neutrophil migration after treatments with SP@Gel/HA and Gel/HA. Untreated group serves as a control. e Schematic of macrophage polarization after treatment with LPS, IL-10, or IL-10/LPS. f Representative fluorescence images of macrophages after different treatment. Red: iNOS (M1 marker); Green: CD163 (M2c marker); Blue: DAPI (nuclear staining). g Schematic of macrophage efferocytosis test toward apoptotic dHL-60 cells under different treatments. h Flow cytometry plots of dHL-60 cells before and after apoptosis induction. i Representative fluorescent images of macrophage efferocytosis toward apoptotic dHL-60 cells under different treatments. Macrophages and apoptotic cells were stained green and red, respectively. All data were generated from at least three independent experiments and presented as the means ± standard deviation. Statistical analysis was performed by one-way ANOVA. ns, not significant; ∗∗∗∗p < 0.0001.

Article Snippet: After another 48 h, macrophage cells were harvested and stained with antibodies against human iNOS (Servicebio, GB11119) and CD163 (Abcam, ab182422) for 30 min. Fluorescence images were acquired using fluorescence microscope.

Techniques: In Vitro, Migration, Comparison, Staining, Control, Fluorescence, Marker, Flow Cytometry, Generated, Standard Deviation

Journal: Bioactive Materials

Article Title: Skin-mimetic bilayer hydrogel normalizes diabetic wound healing by orchestrating inflammatory cell dynamics: An early intervention strategy

doi: 10.1016/j.bioactmat.2026.02.025

Figure Lengend Snippet: Bilayer hydrogel orchestrates inflammatory cell dynamics during the early inflammation phase of diabetic wound healing. a Experimental timeline for assay of early neutrophil recruitment. b Immunohistochemical staining for Ly-6G in wounds at 8 h, 1 d and 3 d after injury. Diabetic wounds were treated with SP/IL-10@Bilayer, SP@Bilayer, IL-10@Bilayer, and saline solution (Model), respectively. Healthy mice treated with saline solution were set as Normal. c Quantitative analysis of Ly-6G + cells in each group. d Relative expression of CXCL-1 on day 1. e Relative expression of MCP-1 on day 1. f Experimental timeline for assay of M1 macrophage infiltration. g Immunofluorescence staining for iNOS in wounds on days 1, 3 and 6 after injury. h Quantitative analysis of iNOS + cells in each group. i-k Relative expressions of macrophage-associated pro-inflammatory cytokines including TNF-α, IL-1β and IL-6 on day 3. l Schematic illustrating the dynamic modulation of inflammatory cells during the early inflammation phase of diabetic wounds by SP/IL-10@Bilayer. All data were generated from at least three independent experiments and presented as the means ± standard deviation. Statistical analysis was performed by one-way ANOVA. # means significant difference compared to the normal group. #p < 0.05, ##p < 0.01 and ###p < 0.001; ∗ means significant difference compared to the model group. ∗p < 0.05; & means significant difference compared to SP/IL-10@Bilayer. & p < 0.05 and && p < 0.01.

Article Snippet: After another 48 h, macrophage cells were harvested and stained with antibodies against human iNOS (Servicebio, GB11119) and CD163 (Abcam, ab182422) for 30 min. Fluorescence images were acquired using fluorescence microscope.

Techniques: Immunohistochemical staining, Staining, Saline, Expressing, Immunofluorescence, Generated, Standard Deviation

Journal: Bioactive Materials

Article Title: Skin-mimetic bilayer hydrogel normalizes diabetic wound healing by orchestrating inflammatory cell dynamics: An early intervention strategy

doi: 10.1016/j.bioactmat.2026.02.025

Figure Lengend Snippet: In vitro assay of inflammation cell modulation under stimulation of SP-loaded Gel/HA and IL-10-loaded Ker/Cu. a Schematic of neutrophil migration test using a transwell system after treatment with the leaching solution of SP@Gel/HA. Gel/HA and blank culture medium were set for comparison. b Wright-Giemsa staining of HL-60 cells before and after differentiation. c Photograph of dHL-60 cells migrating to the lower chamber. d Quantitative analysis of neutrophil migration after treatments with SP@Gel/HA and Gel/HA. Untreated group serves as a control. e Schematic of macrophage polarization after treatment with LPS, IL-10, or IL-10/LPS. f Representative fluorescence images of macrophages after different treatment. Red: iNOS (M1 marker); Green: CD163 (M2c marker); Blue: DAPI (nuclear staining). g Schematic of macrophage efferocytosis test toward apoptotic dHL-60 cells under different treatments. h Flow cytometry plots of dHL-60 cells before and after apoptosis induction. i Representative fluorescent images of macrophage efferocytosis toward apoptotic dHL-60 cells under different treatments. Macrophages and apoptotic cells were stained green and red, respectively. All data were generated from at least three independent experiments and presented as the means ± standard deviation. Statistical analysis was performed by one-way ANOVA. ns, not significant; ∗∗∗∗p < 0.0001.

Article Snippet: The infiltration of pro-inflammatory (M1) macrophages and polarization of M2c macrophages were analyzed by immunofluorescence staining using antibodies against iNOS (Servicebio, GB11119) and CD163 (Servicebio, GB14027), respectively.

Techniques: In Vitro, Migration, Comparison, Staining, Control, Fluorescence, Marker, Flow Cytometry, Generated, Standard Deviation

Journal: Bioactive Materials

Article Title: Skin-mimetic bilayer hydrogel normalizes diabetic wound healing by orchestrating inflammatory cell dynamics: An early intervention strategy

doi: 10.1016/j.bioactmat.2026.02.025

Figure Lengend Snippet: Bilayer hydrogel orchestrates inflammatory cell dynamics during the early inflammation phase of diabetic wound healing. a Experimental timeline for assay of early neutrophil recruitment. b Immunohistochemical staining for Ly-6G in wounds at 8 h, 1 d and 3 d after injury. Diabetic wounds were treated with SP/IL-10@Bilayer, SP@Bilayer, IL-10@Bilayer, and saline solution (Model), respectively. Healthy mice treated with saline solution were set as Normal. c Quantitative analysis of Ly-6G + cells in each group. d Relative expression of CXCL-1 on day 1. e Relative expression of MCP-1 on day 1. f Experimental timeline for assay of M1 macrophage infiltration. g Immunofluorescence staining for iNOS in wounds on days 1, 3 and 6 after injury. h Quantitative analysis of iNOS + cells in each group. i-k Relative expressions of macrophage-associated pro-inflammatory cytokines including TNF-α, IL-1β and IL-6 on day 3. l Schematic illustrating the dynamic modulation of inflammatory cells during the early inflammation phase of diabetic wounds by SP/IL-10@Bilayer. All data were generated from at least three independent experiments and presented as the means ± standard deviation. Statistical analysis was performed by one-way ANOVA. # means significant difference compared to the normal group. #p < 0.05, ##p < 0.01 and ###p < 0.001; ∗ means significant difference compared to the model group. ∗p < 0.05; & means significant difference compared to SP/IL-10@Bilayer. & p < 0.05 and && p < 0.01.

Article Snippet: The infiltration of pro-inflammatory (M1) macrophages and polarization of M2c macrophages were analyzed by immunofluorescence staining using antibodies against iNOS (Servicebio, GB11119) and CD163 (Servicebio, GB14027), respectively.

Techniques: Immunohistochemical staining, Staining, Saline, Expressing, Immunofluorescence, Generated, Standard Deviation

Journal: Materials Today Bio

Article Title: Exosomal lncRNA OTUD6B-AS1 as a pathogenic nanocarrier promotes inflammatory macrophage polarization in endometritis via a targetable ceRNA circuit

doi: 10.1016/j.mtbio.2026.103027

Figure Lengend Snippet: Analysis of macrophage activation in the uterine tissue of cows with endometritis. (A, B) Representative immunofluorescence (IF) staining images (A) and quantitative analysis (B) of iNOS (M1 marker, red) in endometrial tissues from healthy cows and cows with endometritis. Nuclei were counterstained with DAPI (blue). (C, D) Representative IF staining images (C) and quantitative analysis (D) of Arg1 (M2 marker, red) in endometrial tissues. (E, F) Relative mRNA expression levels of iNOS (E) and Arg1 (F) in endometrial tissues, as determined by qPCR. (G, H) Relative expression levels of IL-1β, IL-6 and TNF-α in endometrial tissues, as determined by IHC. (I, J) Relative mRNA expression levels of IL-1β (I) and IL-6 (J) in endometrial tissues, as determined by qPCR. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, ∗∗∗∗p < 0.0001. (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)

Article Snippet: The antibodies used include iNOS (Proteintech, Cat # 18985-1-AP), Arg1 (Proteintech, Cat # 18985-1-AP), Goat Anti-Rabbit IgG (Bioss, Cat # bs-0295G) and Goat Anti-Mouse IgG (Bioss, Cat # bs-0296Gs).

Techniques: Activation Assay, Immunofluorescence, Staining, Marker, Expressing

Journal: Materials Today Bio

Article Title: Exosomal lncRNA OTUD6B-AS1 as a pathogenic nanocarrier promotes inflammatory macrophage polarization in endometritis via a targetable ceRNA circuit

doi: 10.1016/j.mtbio.2026.103027

Figure Lengend Snippet: Exosomes from LPS-stimulated EECs induce pro-inflammatory macrophage activation. (A) Schematic diagram of the experimental setup for exosome uptake. (B) Fluorescence microscopy images showing the uptake of PKH67-labeled exosomes (green) by macrophages. Cytoskeleton was stained with Phalloidin (red), and nuclei were stained with DAPI (blue). (C) Western blotting analysis of phosphorylated NF-κB p65 (p-p65) in macrophages treated with Control-exo or LPS-exo. (D, E) Representative immunofluorescence (IF) staining images (D) and quantitative analysis (E) of iNOS (greed) in macrophages. (F, G) Representative IF staining images (F) and quantitative analysis (G) of Arg1 (red) in macrophages. (H) Schematic diagram of co-culture experiments. (I, J) Relative mRNA expression levels of iNOS (I) and Arg1 (J) in macrophages after co-culture with EECs. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, ∗∗∗∗p < 0.0001. (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)

Article Snippet: The antibodies used include iNOS (Proteintech, Cat # 18985-1-AP), Arg1 (Proteintech, Cat # 18985-1-AP), Goat Anti-Rabbit IgG (Bioss, Cat # bs-0295G) and Goat Anti-Mouse IgG (Bioss, Cat # bs-0296Gs).

Techniques: Activation Assay, Fluorescence, Microscopy, Labeling, Staining, Western Blot, Control, Immunofluorescence, Co-Culture Assay, Expressing

Journal: Materials Today Bio

Article Title: Exosomal lncRNA OTUD6B-AS1 as a pathogenic nanocarrier promotes inflammatory macrophage polarization in endometritis via a targetable ceRNA circuit

doi: 10.1016/j.mtbio.2026.103027

Figure Lengend Snippet: Transcriptomic profiling reveals significant enrichment of lncRNA OTUD6B-AS1 in exosomes derived from LPS-stimulated EECs. (A, B) LPS-exo was treated with RNase A alone or in combination with Triton X-100 for 4 h, and then co-incubated with macrophages. Relative expression levels of iNOS (A) and Arg1 (B) in macrophages. (C) Schematic overview of the RNA sequencing and analysis workflow. (D) Volcano plot showing differentially expressed lncRNAs in LPS-exo compared to Control-exo. (E, F) Gene Ontology (GO) biological process enrichment analysis (E) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis (F) of the differentially expressed lncRNAs. (G) qPCR validation of the 6 upregulated lncRNAs in Control-exo and LPS-exo. (H) LPS-exo was treated with RNase A alone or in combination with Triton X-100 for 4 h, and then co-incubated with macrophages. Relative mRNA expression level of lncRNA OTUD6B-AS1 in macrophages. (I) A proposed competing endogenous RNA (ceRNA) network involving lncRNA OTUD6B-AS1, miR-128, and Notch2. (J) Relative mRNA expression level of lncRNA OTUD6B-AS1 in control and LPS-stimulated EECs. (K, L) RNA fluorescence in situ hybridization (RNA-FISH) showing the subcellular localization of lncRNA OTUD6B-AS1 (red) in EECs (K) and its quantitative cytoplasmic/nuclear distribution (L). Nuclei were stained with DAPI (blue). (M – P) Relative mRNA expression levels of lncRNA OTUD6B-AS1 (M − O) and miR-128 (P) in endometrial tissues from healthy cows and cows with endometritis, as determined by qPCR (O, P) and RNA-FISH (M) with quantification (N). (Q) Western blotting analysis of Notch2, RBP-Jκ, and p-p65 protein levels in endometrial tissues. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, ∗∗∗∗p < 0.0001. (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)

Article Snippet: The antibodies used include iNOS (Proteintech, Cat # 18985-1-AP), Arg1 (Proteintech, Cat # 18985-1-AP), Goat Anti-Rabbit IgG (Bioss, Cat # bs-0295G) and Goat Anti-Mouse IgG (Bioss, Cat # bs-0296Gs).

Techniques: Derivative Assay, Incubation, Expressing, RNA Sequencing, Control, Biomarker Discovery, Fluorescence, In Situ Hybridization, Staining, Western Blot

Journal: Materials Today Bio

Article Title: Exosomal lncRNA OTUD6B-AS1 as a pathogenic nanocarrier promotes inflammatory macrophage polarization in endometritis via a targetable ceRNA circuit

doi: 10.1016/j.mtbio.2026.103027

Figure Lengend Snippet: EECs-derived exosomes induce pro-inflammatory macrophage activation via delivery of lncRNA OTUD6B-AS1. (A) Relative mRNA expression level of lncRNA OTUD6B-AS1 in macrophages treated with Control-exo or LPS-exo. (B, C) RNA-FISH images (B) and quantitative analysis (C) showing lncRNA OTUD6B-AS1 (red) transfer to macrophages after co-culture with Control-exo or LPS-exo. Nuclei were stained with DAPI (blue). (D) Relative mRNA expression level of lncRNA OTUD6B-AS1 in macrophages after transfection with lncRNA OTUD6B-AS1 overexpression plasmids (OE-lncRNA) or control plasmids (OE-NC). ( E – I) Western blotting analysis of Notch2, RBP-Jκ, and p-p65 (E), along with immunofluorescence (IF) quantitative analysis of iNOS (F, G) and Arg1 (H, I) protein levels in macrophages after transfection with OE-lncRNA or OE-NC. (J – N) Western blotting analysis of Notch2, RBP-Jκ, and p-p65 (J), along with IF quantitative analysis of iNOS (K, L) and Arg1 (M, N) protein levels in macrophages after lncRNA OTUD6B-AS1 knockdown (si-lncRNA) or control treatment (si-NC). (O) Relative mRNA expression level of lncRNA OTUD6B-AS1 in exosomes isolated from lncRNA OTUD6B-AS1-knockdown LPS-stimulated EECs (si-lncRNA-LPS-exo) or exosomes from siRNA NC-transfected LPS-stimulated EECs (si-NC-LPS-exo). (P – S) IF quantitative analysis of iNOS (P, Q) and Arg1 (R, S) protein levels in macrophages treated with si-lncRNA-LPS-exo or si-NC-LPS-exo. (T) Relative mRNA expression levels of iNOS and Arg1 in macrophages treated with exosomes isolated from control EECs overexpressing lncRNA OTUD6B-AS1 (OE-lncRNA-Control-exo) or exosomes from control plasmids-transfected EECs (OE-NC-Control-exo). ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, ∗∗∗∗p < 0.0001. (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)

Article Snippet: The antibodies used include iNOS (Proteintech, Cat # 18985-1-AP), Arg1 (Proteintech, Cat # 18985-1-AP), Goat Anti-Rabbit IgG (Bioss, Cat # bs-0295G) and Goat Anti-Mouse IgG (Bioss, Cat # bs-0296Gs).

Techniques: Derivative Assay, Activation Assay, Expressing, Control, Co-Culture Assay, Staining, Transfection, Over Expression, Western Blot, Immunofluorescence, Knockdown, Isolation

Journal: Materials Today Bio

Article Title: Exosomal lncRNA OTUD6B-AS1 as a pathogenic nanocarrier promotes inflammatory macrophage polarization in endometritis via a targetable ceRNA circuit

doi: 10.1016/j.mtbio.2026.103027

Figure Lengend Snippet: lncRNA OTUD6B-AS1 acts as a ceRNA by sponging miR-128 to facilitate pro-inflammatory macrophage activation. (A) Relative mRNA expression level of miR-128 in macrophages treated with Control-exo or LPS-exo. (B) Luciferase reporter assay in HEK293T cells co-transfected with wild-type (WT) or mutant (MUT) lncRNA OTUD6B-AS1 reporter plasmids and miR-128 mimic or mimic NC. (C) RNA pull-down detection of the enrichment of miR-128 to lncRNA OTUD6B-AS1. (D) Ago2 RIP assay analysis of the enrichment of lncRNA OTUD6B-AS1 pulled-down from the Ago2 protein. (E) Relative mRNA expression level of miR-128 in macrophages transfected with OE-NC or OE-lncRNA. (F – J) Western blotting analysis of Notch2, RBP-Jκ, and p-p65 (F), along with immunofluorescence (IF) quantitative analysis of iNOS (G, H) and Arg1 (I, J) protein levels in macrophages co-transfected with OE-lncRNA and miR-128 mimic or mimic NC. (K, L) Relative mRNA expression levels of IL-1β (K) and IL-6 (L) in macrophages co-transfected with OE-lncRNA and miR-128 mimic or mimic NC. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, ∗∗∗∗p < 0.0001.

Article Snippet: The antibodies used include iNOS (Proteintech, Cat # 18985-1-AP), Arg1 (Proteintech, Cat # 18985-1-AP), Goat Anti-Rabbit IgG (Bioss, Cat # bs-0295G) and Goat Anti-Mouse IgG (Bioss, Cat # bs-0296Gs).

Techniques: Activation Assay, Expressing, Control, Luciferase, Reporter Assay, Transfection, Mutagenesis, Western Blot, Immunofluorescence

Journal: Materials Today Bio

Article Title: Exosomal lncRNA OTUD6B-AS1 as a pathogenic nanocarrier promotes inflammatory macrophage polarization in endometritis via a targetable ceRNA circuit

doi: 10.1016/j.mtbio.2026.103027

Figure Lengend Snippet: Notch2 mediates the regulatory effect of the lncRNA OTUD6B-AS1/miR-128 axis on macrophage activation. (A) Predictive analysis of miR-128 targets using multiple databases. (B) Western blotting analysis of Notch2 protein levels in macrophages treated with Control-exo or LPS-exo. (C) Western blotting analysis of Notch2 protein levels in macrophages transfected with OE-NC or OE-lncRNA. (D – H) Western blotting analysis of Notch2, RBP-Jκ, and p-p65 (D), along with immunofluorescence (IF) quantitative analysis of iNOS (E, F) and Arg1 (G, H) protein levels in macrophages treated with OE-NC or OE-lncRNA and the Notch2 inhibitor DAPT. (I) Luciferase reporter assay in HEK293T cells co-transfected with WT or MUT Notch2 3′UTR reporter plasmids and miR-128 mimic or mimic NC. (J, K) Relative protein (J) and mRNA (K) expression levels of Notch2 in macrophages transfected with miR-128 mimic or mimic NC. (L – P) Western blotting analysis of Notch2, RBP-Jκ, and p-p65 (L), along with IF quantitative analysis of iNOS (M, N) and Arg1 (O, P) protein levels in macrophages co-treated with miR-128 inhibitor or inhibitor NC and DAPT. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, ∗∗∗∗p < 0.0001.

Article Snippet: The antibodies used include iNOS (Proteintech, Cat # 18985-1-AP), Arg1 (Proteintech, Cat # 18985-1-AP), Goat Anti-Rabbit IgG (Bioss, Cat # bs-0295G) and Goat Anti-Mouse IgG (Bioss, Cat # bs-0296Gs).

Techniques: Activation Assay, Western Blot, Control, Transfection, Immunofluorescence, Luciferase, Reporter Assay, Expressing

Journal: Materials Today Bio

Article Title: Exosomal lncRNA OTUD6B-AS1 as a pathogenic nanocarrier promotes inflammatory macrophage polarization in endometritis via a targetable ceRNA circuit

doi: 10.1016/j.mtbio.2026.103027

Figure Lengend Snippet: Analysis of macrophage activation in the uterine tissue of cows with endometritis. (A, B) Representative immunofluorescence (IF) staining images (A) and quantitative analysis (B) of iNOS (M1 marker, red) in endometrial tissues from healthy cows and cows with endometritis. Nuclei were counterstained with DAPI (blue). (C, D) Representative IF staining images (C) and quantitative analysis (D) of Arg1 (M2 marker, red) in endometrial tissues. (E, F) Relative mRNA expression levels of iNOS (E) and Arg1 (F) in endometrial tissues, as determined by qPCR. (G, H) Relative expression levels of IL-1β, IL-6 and TNF-α in endometrial tissues, as determined by IHC. (I, J) Relative mRNA expression levels of IL-1β (I) and IL-6 (J) in endometrial tissues, as determined by qPCR. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, ∗∗∗∗p < 0.0001. (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)

Article Snippet: The antibodies used include iNOS (Proteintech, Cat # 18985-1-AP), Arg1 (Proteintech, Cat # 18985-1-AP), Goat Anti-Rabbit IgG (Bioss, Cat # bs-0295G) and Goat Anti-Mouse IgG (Bioss, Cat # bs-0296Gs).

Techniques: Activation Assay, Immunofluorescence, Staining, Marker, Expressing

Journal: Materials Today Bio

Article Title: Exosomal lncRNA OTUD6B-AS1 as a pathogenic nanocarrier promotes inflammatory macrophage polarization in endometritis via a targetable ceRNA circuit

doi: 10.1016/j.mtbio.2026.103027

Figure Lengend Snippet: Exosomes from LPS-stimulated EECs induce pro-inflammatory macrophage activation. (A) Schematic diagram of the experimental setup for exosome uptake. (B) Fluorescence microscopy images showing the uptake of PKH67-labeled exosomes (green) by macrophages. Cytoskeleton was stained with Phalloidin (red), and nuclei were stained with DAPI (blue). (C) Western blotting analysis of phosphorylated NF-κB p65 (p-p65) in macrophages treated with Control-exo or LPS-exo. (D, E) Representative immunofluorescence (IF) staining images (D) and quantitative analysis (E) of iNOS (greed) in macrophages. (F, G) Representative IF staining images (F) and quantitative analysis (G) of Arg1 (red) in macrophages. (H) Schematic diagram of co-culture experiments. (I, J) Relative mRNA expression levels of iNOS (I) and Arg1 (J) in macrophages after co-culture with EECs. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, ∗∗∗∗p < 0.0001. (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)

Article Snippet: The antibodies used include iNOS (Proteintech, Cat # 18985-1-AP), Arg1 (Proteintech, Cat # 18985-1-AP), Goat Anti-Rabbit IgG (Bioss, Cat # bs-0295G) and Goat Anti-Mouse IgG (Bioss, Cat # bs-0296Gs).

Techniques: Activation Assay, Fluorescence, Microscopy, Labeling, Staining, Western Blot, Control, Immunofluorescence, Co-Culture Assay, Expressing

Journal: Materials Today Bio

Article Title: Exosomal lncRNA OTUD6B-AS1 as a pathogenic nanocarrier promotes inflammatory macrophage polarization in endometritis via a targetable ceRNA circuit

doi: 10.1016/j.mtbio.2026.103027

Figure Lengend Snippet: Transcriptomic profiling reveals significant enrichment of lncRNA OTUD6B-AS1 in exosomes derived from LPS-stimulated EECs. (A, B) LPS-exo was treated with RNase A alone or in combination with Triton X-100 for 4 h, and then co-incubated with macrophages. Relative expression levels of iNOS (A) and Arg1 (B) in macrophages. (C) Schematic overview of the RNA sequencing and analysis workflow. (D) Volcano plot showing differentially expressed lncRNAs in LPS-exo compared to Control-exo. (E, F) Gene Ontology (GO) biological process enrichment analysis (E) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis (F) of the differentially expressed lncRNAs. (G) qPCR validation of the 6 upregulated lncRNAs in Control-exo and LPS-exo. (H) LPS-exo was treated with RNase A alone or in combination with Triton X-100 for 4 h, and then co-incubated with macrophages. Relative mRNA expression level of lncRNA OTUD6B-AS1 in macrophages. (I) A proposed competing endogenous RNA (ceRNA) network involving lncRNA OTUD6B-AS1, miR-128, and Notch2. (J) Relative mRNA expression level of lncRNA OTUD6B-AS1 in control and LPS-stimulated EECs. (K, L) RNA fluorescence in situ hybridization (RNA-FISH) showing the subcellular localization of lncRNA OTUD6B-AS1 (red) in EECs (K) and its quantitative cytoplasmic/nuclear distribution (L). Nuclei were stained with DAPI (blue). (M – P) Relative mRNA expression levels of lncRNA OTUD6B-AS1 (M − O) and miR-128 (P) in endometrial tissues from healthy cows and cows with endometritis, as determined by qPCR (O, P) and RNA-FISH (M) with quantification (N). (Q) Western blotting analysis of Notch2, RBP-Jκ, and p-p65 protein levels in endometrial tissues. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, ∗∗∗∗p < 0.0001. (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)

Article Snippet: The antibodies used include iNOS (Proteintech, Cat # 18985-1-AP), Arg1 (Proteintech, Cat # 18985-1-AP), Goat Anti-Rabbit IgG (Bioss, Cat # bs-0295G) and Goat Anti-Mouse IgG (Bioss, Cat # bs-0296Gs).

Techniques: Derivative Assay, Incubation, Expressing, RNA Sequencing, Control, Biomarker Discovery, Fluorescence, In Situ Hybridization, Staining, Western Blot

Journal: Materials Today Bio

Article Title: Exosomal lncRNA OTUD6B-AS1 as a pathogenic nanocarrier promotes inflammatory macrophage polarization in endometritis via a targetable ceRNA circuit

doi: 10.1016/j.mtbio.2026.103027

Figure Lengend Snippet: EECs-derived exosomes induce pro-inflammatory macrophage activation via delivery of lncRNA OTUD6B-AS1. (A) Relative mRNA expression level of lncRNA OTUD6B-AS1 in macrophages treated with Control-exo or LPS-exo. (B, C) RNA-FISH images (B) and quantitative analysis (C) showing lncRNA OTUD6B-AS1 (red) transfer to macrophages after co-culture with Control-exo or LPS-exo. Nuclei were stained with DAPI (blue). (D) Relative mRNA expression level of lncRNA OTUD6B-AS1 in macrophages after transfection with lncRNA OTUD6B-AS1 overexpression plasmids (OE-lncRNA) or control plasmids (OE-NC). ( E – I) Western blotting analysis of Notch2, RBP-Jκ, and p-p65 (E), along with immunofluorescence (IF) quantitative analysis of iNOS (F, G) and Arg1 (H, I) protein levels in macrophages after transfection with OE-lncRNA or OE-NC. (J – N) Western blotting analysis of Notch2, RBP-Jκ, and p-p65 (J), along with IF quantitative analysis of iNOS (K, L) and Arg1 (M, N) protein levels in macrophages after lncRNA OTUD6B-AS1 knockdown (si-lncRNA) or control treatment (si-NC). (O) Relative mRNA expression level of lncRNA OTUD6B-AS1 in exosomes isolated from lncRNA OTUD6B-AS1-knockdown LPS-stimulated EECs (si-lncRNA-LPS-exo) or exosomes from siRNA NC-transfected LPS-stimulated EECs (si-NC-LPS-exo). (P – S) IF quantitative analysis of iNOS (P, Q) and Arg1 (R, S) protein levels in macrophages treated with si-lncRNA-LPS-exo or si-NC-LPS-exo. (T) Relative mRNA expression levels of iNOS and Arg1 in macrophages treated with exosomes isolated from control EECs overexpressing lncRNA OTUD6B-AS1 (OE-lncRNA-Control-exo) or exosomes from control plasmids-transfected EECs (OE-NC-Control-exo). ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, ∗∗∗∗p < 0.0001. (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)

Article Snippet: The antibodies used include iNOS (Proteintech, Cat # 18985-1-AP), Arg1 (Proteintech, Cat # 18985-1-AP), Goat Anti-Rabbit IgG (Bioss, Cat # bs-0295G) and Goat Anti-Mouse IgG (Bioss, Cat # bs-0296Gs).

Techniques: Derivative Assay, Activation Assay, Expressing, Control, Co-Culture Assay, Staining, Transfection, Over Expression, Western Blot, Immunofluorescence, Knockdown, Isolation

Journal: Materials Today Bio

Article Title: Exosomal lncRNA OTUD6B-AS1 as a pathogenic nanocarrier promotes inflammatory macrophage polarization in endometritis via a targetable ceRNA circuit

doi: 10.1016/j.mtbio.2026.103027

Figure Lengend Snippet: lncRNA OTUD6B-AS1 acts as a ceRNA by sponging miR-128 to facilitate pro-inflammatory macrophage activation. (A) Relative mRNA expression level of miR-128 in macrophages treated with Control-exo or LPS-exo. (B) Luciferase reporter assay in HEK293T cells co-transfected with wild-type (WT) or mutant (MUT) lncRNA OTUD6B-AS1 reporter plasmids and miR-128 mimic or mimic NC. (C) RNA pull-down detection of the enrichment of miR-128 to lncRNA OTUD6B-AS1. (D) Ago2 RIP assay analysis of the enrichment of lncRNA OTUD6B-AS1 pulled-down from the Ago2 protein. (E) Relative mRNA expression level of miR-128 in macrophages transfected with OE-NC or OE-lncRNA. (F – J) Western blotting analysis of Notch2, RBP-Jκ, and p-p65 (F), along with immunofluorescence (IF) quantitative analysis of iNOS (G, H) and Arg1 (I, J) protein levels in macrophages co-transfected with OE-lncRNA and miR-128 mimic or mimic NC. (K, L) Relative mRNA expression levels of IL-1β (K) and IL-6 (L) in macrophages co-transfected with OE-lncRNA and miR-128 mimic or mimic NC. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, ∗∗∗∗p < 0.0001.

Article Snippet: The antibodies used include iNOS (Proteintech, Cat # 18985-1-AP), Arg1 (Proteintech, Cat # 18985-1-AP), Goat Anti-Rabbit IgG (Bioss, Cat # bs-0295G) and Goat Anti-Mouse IgG (Bioss, Cat # bs-0296Gs).

Techniques: Activation Assay, Expressing, Control, Luciferase, Reporter Assay, Transfection, Mutagenesis, Western Blot, Immunofluorescence

Journal: Materials Today Bio

Article Title: Exosomal lncRNA OTUD6B-AS1 as a pathogenic nanocarrier promotes inflammatory macrophage polarization in endometritis via a targetable ceRNA circuit

doi: 10.1016/j.mtbio.2026.103027

Figure Lengend Snippet: Notch2 mediates the regulatory effect of the lncRNA OTUD6B-AS1/miR-128 axis on macrophage activation. (A) Predictive analysis of miR-128 targets using multiple databases. (B) Western blotting analysis of Notch2 protein levels in macrophages treated with Control-exo or LPS-exo. (C) Western blotting analysis of Notch2 protein levels in macrophages transfected with OE-NC or OE-lncRNA. (D – H) Western blotting analysis of Notch2, RBP-Jκ, and p-p65 (D), along with immunofluorescence (IF) quantitative analysis of iNOS (E, F) and Arg1 (G, H) protein levels in macrophages treated with OE-NC or OE-lncRNA and the Notch2 inhibitor DAPT. (I) Luciferase reporter assay in HEK293T cells co-transfected with WT or MUT Notch2 3′UTR reporter plasmids and miR-128 mimic or mimic NC. (J, K) Relative protein (J) and mRNA (K) expression levels of Notch2 in macrophages transfected with miR-128 mimic or mimic NC. (L – P) Western blotting analysis of Notch2, RBP-Jκ, and p-p65 (L), along with IF quantitative analysis of iNOS (M, N) and Arg1 (O, P) protein levels in macrophages co-treated with miR-128 inhibitor or inhibitor NC and DAPT. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, ∗∗∗∗p < 0.0001.

Article Snippet: The antibodies used include iNOS (Proteintech, Cat # 18985-1-AP), Arg1 (Proteintech, Cat # 18985-1-AP), Goat Anti-Rabbit IgG (Bioss, Cat # bs-0295G) and Goat Anti-Mouse IgG (Bioss, Cat # bs-0296Gs).

Techniques: Activation Assay, Western Blot, Control, Transfection, Immunofluorescence, Luciferase, Reporter Assay, Expressing

Journal: International Journal of Pharmaceutics: X

Article Title: ROS-triggered smart hydrogel adhesives for modulating the inflammatory microenvironment and promoting tendon-bone interface regeneration

doi: 10.1016/j.ijpx.2026.100502

Figure Lengend Snippet: Immunohistochemical analysis of (A) Col-I and (B) Col-III. (C) Quantitative analysis of the Col-I/Col-III Ratio in tissue sections. (D) Immunofluorescence double staining analysis of iNOS and CD206. Quantitative Analysis of (E) iNOS and (F) CD206 in tissue sections. (G) Force-displacement curve, (H) failure load, and (I) stiffness of biomechanical testing for tendon-bone complexes. n = 6, Data are presented as mean ± standard deviation (SD).

Article Snippet: Meanwhile, immunofluorescence (IF) staining was conducted to detect the expression of M1 macrophage marker and M2 macrophage marker using Recombinant Anti-iNOS antibody (Rabbit mAb) (Servicebio, GB153965 , 1:2000) and Anti-Mannose Receptor/CD206 Rabbit pAb (Servicebio, GB113497 , 1:800), respectively.

Techniques: Immunohistochemical staining, Immunofluorescence, Double Staining, Standard Deviation