Journal: Bioactive Materials

Article Title: 3D-MSCs apoptotic bodies-integrated conductive hydrogel mitigates spinal cord injury via immunoregulation and alleviating neuronal pyroptosis

doi: 10.1016/j.bioactmat.2026.01.043

Figure Lengend Snippet: Preparation and characterization of 3D-ABs and Mxene nanosheets. (A) Graphical illustration of the 3D-ABs isolation procedure. (B) 2D-BMSCs and 3D-BMSCs stained by CD29 and CD44 (scale bar, 40 μm). (C) SEM images of 3D-ABs (scale bar, 1 μm). (D) 3D-ABs stained by CIQC (scale bar, 10 μm). (E) DLS analysis for the 3D-ABs. (F) Expression of biomarkers of 3D-ABs and Hydrogel-3D-ABs including C1QC, C3B, H2B, and H3, β-actin was utilized as a loading control. (G) FCM analysis of the percentage of Dil-positive PC12 and BV2 cells after treatment with DiI-labelled 3D-ABs. (H) Uptake of Dil-labelled 3D-ABs and Hydrogel-3D-ABs by PC12 and BV2 cells (scale bar, 20 μm). (I) Frozen sections of DiI-labelled 3D-ABs and Hydrogel-3D-ABs treated spinal cord were stained for Neun and CD68 (scale bars, 20 μm). (J) SEM and EDS elemental mapping images of the Mxene nanosheets (scale bar, 20 μm). (K) Live/dead cell staining images for PC12 and BV2 cells after different treatments (scale bar, 200 μm).

Article Snippet: BV2 microglia (CL-0493) were obtained from Procell Life Science & Technology Co., Ltd.

Techniques: Isolation, Staining, Expressing, Control

Journal: Bioactive Materials

Article Title: 3D-MSCs apoptotic bodies-integrated conductive hydrogel mitigates spinal cord injury via immunoregulation and alleviating neuronal pyroptosis

doi: 10.1016/j.bioactmat.2026.01.043

Figure Lengend Snippet: Composite hydrogel inhibits oxidative damage in vitro and in vivo. (A) DCFH-DA staining of PC12 and BV2 cells treated with different hydrogel under TBHP treatment (scale bar, 200 μm). (B) Frozen sections of different hydrogel-treated spinal cords were stained for DHE (scale bar: 50 μm). (C, D) DCFH-DA integrated intensity analysis of PC12 and BV2 cells (n = 3). (E) Analysis and quantification of DHE integrated intensity in each group (n = 3). (F) Transcriptome GSEA analysis. The data are presented as the means ± SEMs (n = 3); ∗p < 0.05, indicates significant differences; ns, is not significant. Statistical analysis was performed using two-way ANOVA followed by Tukey's multiple comparison test.

Article Snippet: BV2 microglia (CL-0493) were obtained from Procell Life Science & Technology Co., Ltd.

Techniques: In Vitro, In Vivo, Staining, Comparison

Journal: Bioactive Materials

Article Title: 3D-MSCs apoptotic bodies-integrated conductive hydrogel mitigates spinal cord injury via immunoregulation and alleviating neuronal pyroptosis

doi: 10.1016/j.bioactmat.2026.01.043

Figure Lengend Snippet: Composite hydrogel promotes the polarization of BV2 cells to M2 types and alleviates PC12 cell pyroptosis in vitro inflammatory environment. (A) Representative western blots showing protein expression of iNOS and Arg-1 in each group, β-actin was utilized as a loading control. (B) Quantitative analysis of relative expression of iNOS and Arg-1. (C) Representative immunofluorescence images of CD68 positive and iNOS positive BV2 cells (scale bar: 20 μm). (D) Representative immunofluorescence images of CD68 positive and Arg-1 positive BV2 cells (scale bar: 20 μm). (E, F) Quantitative analysis of relative fluorescence intensity of iNOS and Arg-1. (G) Representative western blots showing the expression of NLRP3, Caspase-1, IL-1β, ASC, GSDMD-N, and IL-18 protein associated with pyroptosis, β-actin was utilized as a loading control. (H) PI staining of PC12 cells in each group (scale bar, 100 μm). (I) Quantitative analysis of PI staining of PC12 cells (n = 3). (J) Quantitative analysis of relative expression of NLRP3, Caspase-1, IL-1β, ASC, GSDMD-N and IL-18 (n = 3). The data are presented as the means ± SEMs (n = 3); ∗p < 0.05, indicates significant differences; ns, is not significant. Statistical analysis was performed using two-way ANOVA followed by Tukey's multiple comparison test.

Article Snippet: BV2 microglia (CL-0493) were obtained from Procell Life Science & Technology Co., Ltd.

Techniques: In Vitro, Western Blot, Expressing, Control, Immunofluorescence, Fluorescence, Staining, Comparison

Journal: Bioactive Materials

Article Title: M2 macrophage-derived exosomes delivering haptoglobin and interleukin-10 plasmids for synergistic therapy of intracerebral hemorrhage

doi: 10.1016/j.bioactmat.2026.01.047

Figure Lengend Snippet: Schematic illustration of the preparation of M2-exo@HI and its mediated therapeutic mechanisms and signaling pathways. (a) RAW264.7 macrophages were polarized to M2 phenotype using DEX, followed by M2-exo isolation from cell supernatant via differential centrifugation. M2-exo@HI was prepared through encapsulating HI into M2-exo by electroporation. (b) M2-exo@HI crossed the BBB and localized to microglia in the hemorrhagic brain, delivering the HI plasmid into the nucleus. This prompted expression and secretion of Hp and IL-10 by microglia. The released Hp inhibited Hb toxicity by binding to Hb. IL-10 shifted microglial polarization from the pro-inflammatory M1 phenotype toward the reparative M2 phenotype, removing hematoma by engulfing erythrocyte and Hb-Hp complex, and decreasing pro-inflammatory cytokine levels—collectively enhancing neuroprotection and BBB repair. These beneficial outcomes were linked to the inhibition of the IL-17 and NF-κB signaling pathways and activation of the PI3K-Akt and ferroptosis pathways.

Article Snippet: For CLSM observation, 100 μL ICG and M2-exo@ICG were incubated with M1 microglia for 0.25 h, 0.5 h, 1 h and 2 h. Then cells were washed, fixed, stained with DAPI and observed by CLSM (A1R+, Nikon, Japan).

Techniques: Protein-Protein interactions, Isolation, Centrifugation, Electroporation, Plasmid Preparation, Expressing, Binding Assay, Inhibition, Activation Assay

Journal: Bioactive Materials

Article Title: M2 macrophage-derived exosomes delivering haptoglobin and interleukin-10 plasmids for synergistic therapy of intracerebral hemorrhage

doi: 10.1016/j.bioactmat.2026.01.047

Figure Lengend Snippet: Validation of M2-exo targeting, Hp/IL-10 transfection expression, and Hp/Hb binding. (a) Fluorescence imaging showing cellular uptake of ICG and M2-exo@ICG by M1 microglia. (b,c) Flow cytometry and corresponding quantification of RhB and M2-exo@RhB internalized by M1 microglia (n = 3). (d,e) Schematic illustration and quantitative analysis of the in vitro phagocytosis-release kinetics of M2-exo@RhB in BV2 under ICH-mimicking stimulation (n = 6). (f) Fluorescence images showing Hp and IL-10 expression in M1 microglia treated with M2-exo@HI for 12, 24, 48, 72 h. (g) Mean fluorescence intensity (MFI) quantification of Hp and IL-10 expression (n = 3). (h,i) ELISA measurements of secreted Hp and IL-10 protein levels (n = 3). (j,k) qPCR analysis of relative Hp and IL-10 mRNA expression (n = 3). (l) Western blot detection of Hp and IL-10 protein expression. (m) Densitometric quantification of Hp and IL-10 protein levels from Western blot (n = 3). (n) Co-immunoprecipitation assay confirming the formation of Hp-Hb complex. Data are presented as mean ± SD. Statistical significance was calculated by unpaired Student's t -test (c and e), and one-way ANOVA with Tukey's multiple comparisons test (g-k and m).

Article Snippet: For CLSM observation, 100 μL ICG and M2-exo@ICG were incubated with M1 microglia for 0.25 h, 0.5 h, 1 h and 2 h. Then cells were washed, fixed, stained with DAPI and observed by CLSM (A1R+, Nikon, Japan).

Techniques: Biomarker Discovery, Transfection, Expressing, Binding Assay, Fluorescence, Imaging, Flow Cytometry, In Vitro, Enzyme-linked Immunosorbent Assay, Western Blot, Co-Immunoprecipitation Assay

Journal: Bioactive Materials

Article Title: M2 macrophage-derived exosomes delivering haptoglobin and interleukin-10 plasmids for synergistic therapy of intracerebral hemorrhage

doi: 10.1016/j.bioactmat.2026.01.047

Figure Lengend Snippet: M2-exo@HI promotes in vitro microglia polarization, BBB repair and neuroprotection. (a) Flow cytometry analysis of M1-type (CD86 + ) and M2-type microglia (CD163 + ) following treatment with different formulations. (b,c) Percentages of CD86 + and CD163 + microglia populations (n = 3). (d–g) The cytokine levels of IL-10, TGF-β, TNF-α, and IL-1β in treated microglia (n = 3). (h) Fluorescence microscopy images showing erythrophagocytosis by microglia across treatment groups. (i) Schematic of the in vitro BBB model assessing FITC-dextran permeability using a transwell assay. (j) Quantitative analysis of FITC-dextran penetration (n = 7). (k) Flow cytometry analysis of neuronal apoptosis across treatments (n = 3). (l) Quantitative analysis of neuronal apoptosis (n = 3). Data are presented as mean ± SD. Statistical significance was tested by one-way ANOVA with Tukey's multiple comparisons test.

Article Snippet: For CLSM observation, 100 μL ICG and M2-exo@ICG were incubated with M1 microglia for 0.25 h, 0.5 h, 1 h and 2 h. Then cells were washed, fixed, stained with DAPI and observed by CLSM (A1R+, Nikon, Japan).

Techniques: In Vitro, Flow Cytometry, Fluorescence, Microscopy, Permeability, Transwell Assay

Journal: Bioactive Materials

Article Title: M2 macrophage-derived exosomes delivering haptoglobin and interleukin-10 plasmids for synergistic therapy of intracerebral hemorrhage

doi: 10.1016/j.bioactmat.2026.01.047

Figure Lengend Snippet: In vivo therapeutic effects of M2-exo@HI in hemorrhagic stroke. (a) The schematic diagram illustrates the construction of mouse cerebral hemorrhage model and treatment regimens. (b) Digital photos showing cerebral hematoma of ICH mice in different groups. (c) Quantitative measurements of hemoglobin concentration in different groups (n = 3). (d) Cerebral edema quantification by brain water content measurements (n = 3). (e) CLSM images showing M1 microglia (CD86 + , green) and M2 microglia (CD163 + , red) in different groups. Nucleus were stained with DAPI (blue). (f) Immunofluorescence staining showing co-localization of Hb/Hp with microglia in different groups. (g) Representative images of HE, Nissl, and TUNEL staining. Data are presented as mean ± SD. Statistical significance was tested by one-way ANOVA with Tukey's multiple comparisons test.

Article Snippet: For CLSM observation, 100 μL ICG and M2-exo@ICG were incubated with M1 microglia for 0.25 h, 0.5 h, 1 h and 2 h. Then cells were washed, fixed, stained with DAPI and observed by CLSM (A1R+, Nikon, Japan).

Techniques: In Vivo, Concentration Assay, Staining, Immunofluorescence, TUNEL Assay

Journal: Frontiers in Immunology

Article Title: miR-511-3p dysregulation-mediated AKT3/USP8 signaling imbalance: a molecular bridge between neuroinflammation and PSCI

doi: 10.3389/fimmu.2026.1766326

Figure Lengend Snippet: Regulatory effects of miR-511-3p on the AKT3/USP8 in BV2 cells. (A–C) . Relative expression of miR-511-3p/AKT3/USP8 in BV2 cell model. (D) . miR-511-3p mimic promoted miR-511-3p expression. (E) . pcDNA3.1-AKT3 promoted AKT3 expression, but miR-511-3p mimic inhibited AKT3 expression, while this inhibition was rescued by pcDNA3.1-AKT3. (F) . pcDNA3.1-AKT3 inhibited USP8 expression, but miR-511-3p mimic promoted USP8 expression, while this promotion was rescued by pcDNA3.1-AKT3. ** P < 0.01, *** P < 0.001, compared to mimic NC; ## P < 0.01, compared to pcDNA3.1; & P < 0.05, &&& P < 0.001, compared to miR-511-3p mimic.

Article Snippet: The mouse microglia cell line BV2 cells (Procell, China, CL-0697) were resuscitated, seeded in six-well plates, added with DMEM containing 10% fetal bovine serum (FBS).

Techniques: Expressing, Inhibition

Journal: Frontiers in Immunology

Article Title: miR-511-3p dysregulation-mediated AKT3/USP8 signaling imbalance: a molecular bridge between neuroinflammation and PSCI

doi: 10.3389/fimmu.2026.1766326

Figure Lengend Snippet: miR-511-3p inhibited inflammation by targeting AKT3/USP8 in the cell model. (A) The expression level of IL-1β was regulatory by miR-511-3p/AKT3/USP8. (B) The expression level of IL-6 was regulatory by miR-511-3p/AKT3/USP8. (C) The expression level of TNF-α was regulatory by miR-511-3p/AKT3/USP8. *** P < 0.001, compared to BV2; ## P < 0.01, ### P < 0.001, compared to BV2+OGD/R; & P < 0.05, && P < 0.01, &&& P < 0.001, compared to BV2+OGD/R+miR-511-3p mimic.

Article Snippet: The mouse microglia cell line BV2 cells (Procell, China, CL-0697) were resuscitated, seeded in six-well plates, added with DMEM containing 10% fetal bovine serum (FBS).

Techniques: Expressing