Journal: Materials Today Bio

Article Title: Toll-like receptor 9-overexpressing red blood cell biomimetic microbubbles adsorb cell-free DNA to relieve steatotic liver ischemia-reperfusion injury

doi: 10.1016/j.mtbio.2026.103558

Figure Lengend Snippet: RBC-MBs promotes the polarization of macrophages towards the M2 phenotype by inhibiting the TLR9-MyD88 and cGAS-STING pathways (A-F) RBC-MB treatment significantly attenuated CpG DNA-induced upregulation of pro-inflammatory cytokines (A-B), key proteins in the TLR9-MyD88 (C-D) and cGAS-STING (E-F) pathways. RAW264.7 cells were treated with 1 μg mL -1 CpG DNA for 24 h, followed by incubation with 0.5 mL RBC-MBs (concentration: 1 × 10 8 particles mL -1 ) for 24 h, and then subjected to western blot analysis (A, C, E). IL-1α, TNF-α, IL-6, TLR9, MyD88, STING, and cGAS levels were normalized to the β-actin level in each sample. p-NF-κB, p-TBK1, and p-IRF3 levels were normalized to their respective total protein levels (NF-κB, TBK1, and IRF3) in each sample. These normalized values were then used for statistical analysis (B, D, F), n = 6. (G) RBC-MB treatment significantly attenuated CpG DNA-induced upregulation of CD86 in macrophages. RAW264.7 cells were treated with 1 μg mL -1 CpG DNA for 24 h, followed by incubation with 0.5 mL RBC-MBs (concentration: 1 × 10 8 particles mL -1 ) for 24 h, then immunostained with an anti-CD86 antibody (green) and DAPI (blue, nuclei), and imaged by fluorescence microscopy. Representative images (left panel) and quantitative fluorescence graph (right panel) are shown. Scale bar: 50 μm, n = 6. (H-J) RBC-MB treatment significantly inhibited hepatic IRI-induced polarization of macrophages toward the M1 phenotype (H) and activation of the TLR9-MyD88 and cGAS-STING pathways (I-J). MASLD rats underwent 45 min of hepatic ischemia followed by reperfusion. At 0, 6, 12, 18, and 24 h post-reperfusion, the rats were treated with either Con-MBs or RBC-MBs. Primary macrophages were isolated from rat livers, then co-stained with CD68 and CD163 followed by flow cytometric analysis (H), n = 3/4, or subjected to western blotting (I-J). CD206, iNOS, TLR9, MyD88, STING, and cGAS levels were normalized to the β-actin level in each sample, and these normalized values were used for statistical analysis (J), n = 3. For (B, D, F, G right panel , H right panel , J), error bars represent mean ± standard deviation. P values were calculated using the unpaired Student's t-test (B, D, F, G right panel ) or one-way analysis of variance (ANOVA) (H right panel , J). *, P < 0.05; **, P < 0.01; ***, P < 0.001; ****, P < 0.0001; ns, not significant.(For interpretation of the references to color in this figure legend, the reader is referred to the Web version of this article.)

Article Snippet: Primary antibodies used included: rabbit polyclonal antibodies against TLR9 (A14642, Abclonal), NF-κB (A2547, Abclonal), MyD88 (A0980, Abclonal), cGAS (A8335, Abclonal), IL-1α (A2170, Abclonal), TNF-α (A0277, Abclonal), IL-6 (A0286, Abclonal), CD206 (A8301, Abclonal), CD86 (A16805, Abclonal), CD47 (A1838, Abclonal), and ATP1A (A0643, Abclonal); rabbit monoclonal antibodies against STING (13647, CST, Beverly, MA, USA), p-NF-κB (3033, CST), p-TBK1 (5483, CST), TBK1 (ab40676, Abcam), p-IκB (2859, CST), IκB (4812, CST), p-IRF3 (4947, CST), and β-actin (AC026, Abclonal).

Techniques: Incubation, Concentration Assay, Western Blot, Fluorescence, Microscopy, Activation Assay, Isolation, Staining, Standard Deviation

Journal: Bioactive Materials

Article Title: Injection site dictates the immune response to a biodegradable polymer and corresponding collagen regeneration

doi: 10.1016/j.bioactmat.2026.04.004

Figure Lengend Snippet: LA released from T-gel degradation induces macrophage M2 polarization and promotes fibroblast activation and collagen production. (A) Flow cytometry analysis of the association between T-gel degradation products and macrophage M2 polarization ( n = 3, data represent mean ± s.d.). ∗∗∗∗P < 0.0001. (B) RNA-seq analysis of M2 macrophage marker expression in tissues one week after ID and SC injection of T-gel. (C–D) Immunofluorescence analysis of CD206 (C) and FAPα (D) expression in tissues ( n = 3, data represent mean ± s.d.). ∗∗P < 0.01. (E) Schematic diagram of the co-culture system showing RAW264.7 macrophages pretreated with T-gel extract medium that were subsequently co-cultured with L929 fibroblasts. (F–G) Collagen expression in L929 cells from the co-culture system assessed by Western blot (F) and ELISA (G). “C” represents RAW264.7 pretreated with blank medium; “T” represents RAW264.7 pretreated with T-gel extract medium. n = 3, data represent mean ± s.d. ∗P < 0.05 and ∗∗P < 0.01. (H) ELISA analysis of TGFβ expression in RAW264.7 cells treated with T-gel extract medium ( n = 3, data represent mean ± s.d.). ∗∗P < 0.01. (I) Schematic representation of LA, released during T-gel degradation, programming macrophages toward an M2 phenotype and subsequently promoting fibroblast activation and collagen secretion.

Article Snippet: Following permeabilization, cells were stained with Phycoerythrin (PE)-conjugated anti-mouse CD206 antibody (Elabscience, E-AB-F1135D) for 30 min at 4 °C in the dark.

Techniques: Activation Assay, Flow Cytometry, RNA Sequencing, Marker, Expressing, Injection, Immunofluorescence, Co-Culture Assay, Cell Culture, Western Blot, Enzyme-linked Immunosorbent Assay

Journal: Bioactive Materials

Article Title: Spatiotemporally engineered microneedle for microenvironment remodeling propels mucosal regeneration after tracheal mucosal injury

doi: 10.1016/j.bioactmat.2026.01.026

Figure Lengend Snippet: Evaluation of mucosal status after tracheal mucosal injury in rabbits . (a) Gross morphology and Masson staining of mucosal defects (n = 3). (b – f) Tracheal samples were collected on Day 10, and RNA sequencing was performed to assess biological differences between Native and Model groups (n = 3). (b) Principal components analysis of samples. (c) Volcano plot of DEGs. (d) KEGG enrichment analysis of DEGs. (e) Chord plot of enriched KEGG terms. (f) Heatmaps of DEGs associated with inflammation and oxidative stress. (g) IF staining of FISH (marker of bacteria, pink), iNOS (marker of M1 macrophages, orange), CD206 (marker of M2 macrophages, green), and immunohistochemical staining of CD31 (marker of endothelial cells) in various samples, blood vessels are denoted by triangles.

Article Snippet: Cells were then incubated overnight at 4 °C with rabbit polyclonal antibodies against CD86 or CD206 (1:200, Proteintech), followed by 1 h incubation with Alexa Fluor 488-conjugated or CoraLite Plus 594-conjugated goat anti-rabbit IgG secondary antibodies (1:200, Proteintech).

Techniques: Staining, RNA Sequencing, Marker, Bacteria, Immunohistochemical staining

Journal: Bioactive Materials

Article Title: Spatiotemporally engineered microneedle for microenvironment remodeling propels mucosal regeneration after tracheal mucosal injury

doi: 10.1016/j.bioactmat.2026.01.026

Figure Lengend Snippet: Effects of Gel-AgNA/MgGA MN on oxidative stress, macrophage polarization, and angiogenesis after treatment with Gel, Gel-MgGA, or Gel-AgNA/MgGA MN. (a) Representative fluorescence images of DCFH-DA staining (marker of ROS, green) and quantitative analysis of ROS levels in RAW264.7 cells (n = 3). (b – c) Immunofluorescence images and quantitative analysis of CD86+ and CD206+ macrophages in RAW264.7 cells after different treatments (n = 3). (d) Flow cytometry analysis of RAW264.7 macrophages (n = 3). (e) Schematic illustration of HUVECs co-cultured with various MN. (f) Representative images and quantification of Transwell assays in HUVECs (n = 3). (g) Representative images and quantification of EdU assays in HUVECs (n = 3). (h) RT-PCR analysis of angiogenesis-related gene expression in HUVECs after co-culture with MN for 3 days (n = 3).

Article Snippet: Cells were then incubated overnight at 4 °C with rabbit polyclonal antibodies against CD86 or CD206 (1:200, Proteintech), followed by 1 h incubation with Alexa Fluor 488-conjugated or CoraLite Plus 594-conjugated goat anti-rabbit IgG secondary antibodies (1:200, Proteintech).

Techniques: Fluorescence, Staining, Marker, Immunofluorescence, Flow Cytometry, Cell Culture, Reverse Transcription Polymerase Chain Reaction, Gene Expression, Co-Culture Assay

Journal: Bioactive Materials

Article Title: Spatiotemporally engineered microneedle for microenvironment remodeling propels mucosal regeneration after tracheal mucosal injury

doi: 10.1016/j.bioactmat.2026.01.026

Figure Lengend Snippet: Gel-AgNA/MgGA MN remodel the microenvironment after TMI by regulating infection, inflammation, oxidative stress, and vascular disruption. (a) FISH (marker of bacteria, red) staining of harvested samples after treated with Gel, Gel-AgNA, Gel-MgGA, and Gel-AgNA/MgGA MN. (b, c) Quantitative analysis of bacterial coverage and fluorescence intensity of FISH (n = 9). (d) Heatmaps of DEGs related to inflammation and oxidative stress. (e) RT-PCR analysis showing the relative expression levels of IL-1, TNF-α, HIF-1α, and IL-6 (n = 3). (f) IF staining of iNOS (marker of M1 macrophages, red) and CD206 (marker of M2 macrophages, green). (g) Quantitative analysis of iNOS and CD206 fluorescence intensity. (h) RT-PCR analysis showing the relative expression levels of Arg1 and CD206. (i) IF staining of CD31 (marker of blood vessels, red). (j, k) RT-PCR analysis showing the relative expression levels of VEGF and HIF-1α (n = 3).

Article Snippet: Cells were then incubated overnight at 4 °C with rabbit polyclonal antibodies against CD86 or CD206 (1:200, Proteintech), followed by 1 h incubation with Alexa Fluor 488-conjugated or CoraLite Plus 594-conjugated goat anti-rabbit IgG secondary antibodies (1:200, Proteintech).

Techniques: Infection, Disruption, Marker, Bacteria, Staining, Fluorescence, Reverse Transcription Polymerase Chain Reaction, Expressing