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: Primary antibodies against NLRP3 (cat. no. 15101) were obtained from Cell Signaling Technology (Beverly, Massachusetts, USA).

Techniques: In Vitro, Western Blot, Expressing, Control, Immunofluorescence, Fluorescence, 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: The composite hydrogel inhibits post-SCI pyroptosis in neurons. (A) Immunofluorescence images of residual neurons existing in the anterior horn of the spinal cord (scale bar, 500 μm and 200 μm). (B) Immunofluorescence image of Caspase-1 expression of neurons 3 days after SCI (scale bar, 20 μm). (C) Quantitative analysis of relative fluorescence intensity of Caspase-1 protein expression in neurons within the specified groups (n = 3). (D) Immunofluorescence image of GSDMD-N expression of neurons 3 days after SCI (scale bar, 20 μm). (E) Quantitative analysis of relative fluorescence intensity of GSDMD-N protein expression in neurons within the specified groups (n = 3). (F) Representative western blots showing the expression of NLRP3, Caspase-1, IL-1β, ASC, GSDMD-N, and IL-18 protein 3 days after SCI, GAPDH was utilized as a loading control. (G) 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: Primary antibodies against NLRP3 (cat. no. 15101) were obtained from Cell Signaling Technology (Beverly, Massachusetts, USA).

Techniques: Immunofluorescence, Expressing, Fluorescence, Western Blot, Control, Comparison

Journal: Bioengineering & Translational Medicine

Article Title: Engineering macrophage phenotype switching via nucleotide‐binding oligomerization domain‐like receptor protein 3 inflammasome inhibition: A translational approach using antibiotic cement for diabetic foot ulcers

doi: 10.1002/btm2.70073

Figure Lengend Snippet: Regulation of NLRP3 inflammasome activation by antibiotic‐loaded bone cement (ALBC). (a) Schematic diagram of NLRP3 inflammasome activation; (b) immunohistochemistry detection of NLRP3 and Caspase‐1 expression in full‐thickness skin tissue surrounding the wound edge in mice; (c, d) western blot analysis of Pro‐Caspase‐1 and cleaved Caspase‐1 expression in cells; (e) IF detection of NLRP3 and apoptosis‐associated speck‐like protein containing a CARD (ASC) co‐localization. N = 6; cell experiments were repeated three times; *** p < 0.001 between groups. ALBC‐H, high‐dose ALBC; ALBC‐L, low‐dose ALBC; ALBC‐M, medium‐dose ALBC; HG, high‐glucose.

Article Snippet: Primary antibodies against NLRP3 (CSB‐ PA209407 , 1:100, Cusabio) and Caspase‐1 (CSB‐ PA001227 , 1:100, Cusabio) were diluted according to the manufacturer's instructions, applied to the slides, and incubated overnight at 4°C.

Techniques: Activation Assay, Immunohistochemistry, Expressing, Western Blot

Journal: Bioengineering & Translational Medicine

Article Title: Engineering macrophage phenotype switching via nucleotide‐binding oligomerization domain‐like receptor protein 3 inflammasome inhibition: A translational approach using antibiotic cement for diabetic foot ulcers

doi: 10.1002/btm2.70073

Figure Lengend Snippet: Role of the NLRP3 inflammasome in the therapeutic effect of antibiotic‐loaded bone cement (ALBC) in diabetic foot ulcer (DFU) mice. (a) Schematic diagram of the treatment protocol in which diabetic foot ulcer (DFU0 mice received high‐dose antibiotic‐loaded bone cement [ALBC‐H] in combination with the NLRP3 activator Nigericin or the inhibitor MCC950); (b) wound healing rates on Days 7 and 14 in each group; (c) immunohistochemistry analysis of NLRP3 and Caspase‐1 expression in skin tissues from each group; (d) enzyme‐linked immunosorbent assay analysis of IL‐1β and IL‐18 cytokine levels; (e) immunofluorescence detection of TNF‐α, iNOS, CD206, and CD163 protein expression in mouse skin tissues. N = 6; * p < 0.05, ** p < 0.01, *** p < 0.001 between groups.

Article Snippet: Primary antibodies against NLRP3 (CSB‐ PA209407 , 1:100, Cusabio) and Caspase‐1 (CSB‐ PA001227 , 1:100, Cusabio) were diluted according to the manufacturer's instructions, applied to the slides, and incubated overnight at 4°C.

Techniques: Immunohistochemistry, Expressing, Enzyme-linked Immunosorbent Assay, Immunofluorescence

Journal: Bioengineering & Translational Medicine

Article Title: Engineering macrophage phenotype switching via nucleotide‐binding oligomerization domain‐like receptor protein 3 inflammasome inhibition: A translational approach using antibiotic cement for diabetic foot ulcers

doi: 10.1002/btm2.70073

Figure Lengend Snippet: Effects of antibiotic‐loaded bone cement on wound tissues of diabetic foot ulcer (DFU) patients. (a) Immunofluorescence detection of M1 and M2 macrophage phenotype markers in DFU patient wound tissues; (b) immunohistochemistry analysis of NLRP3 and Caspase‐1 expression in DFU patient wound tissues. N = 3; *** p < 0.001 between groups.

Article Snippet: Primary antibodies against NLRP3 (CSB‐ PA209407 , 1:100, Cusabio) and Caspase‐1 (CSB‐ PA001227 , 1:100, Cusabio) were diluted according to the manufacturer's instructions, applied to the slides, and incubated overnight at 4°C.

Techniques: Immunofluorescence, Immunohistochemistry, Expressing

Journal: Chinese Medicine

Article Title: Moxibustion ameliorates abnormal subchondral bone remodeling by promoting ACSL1-mediated autophagy to degrade NLRP3 in osteoarthritis

doi: 10.1186/s13020-025-01182-2

Figure Lengend Snippet: Moxibustion inhibits NLRP3 activation in OA subchondral bone. A Schematic of moxibustion inhibiting NLRP3 activation. B Western blotting detection of NLRP3, IL-1β, IL-18, and Caspase-1 proteins in subchondral bone. C , D IHC detection of IL-1β, IL-18 expression in each group. Scale bar: 250 μm; 50 μm. E , F Quantitative analysis of IHC staining of IL-1β, IL-18 in subchondral bone tissues (n = 5 per group). G Co-localization imaging of NLRP3 (red) and Caspase-1 (green). Scale bar: 50 μm; 10 μm. H – J Intensity of NLRP3 and Caspase-1 co-localization in different groups. K – M The expression levels of NLRP3, IL-1β, and Caspase-1 proteins in subchondral bone were detected by western blotting (n = 3 per group). Data presented as the mean ± SD. ns, not significant, * p < 0.05, ** p < 0.01, *** p < 0.001

Article Snippet: Primary antibody against NLRP3 (Proteintech, 68102-1-I; 1:80), Caspase-1 (Abmart, M025280F; 1:150), SQSTM1 (Abmart, T55546F; 1:200), LAMP1 (Proteintech, 21997-1-AP; 1:200), LC3B (Proteintech, 18725-1-AP; 1:200), ATG5 (Abmart, T55766F; 1:100) and PPARγ (ZENBIO, 340844; 1:200) were added to the slices at 4 °C for over 12 h. After being incubated with anti-rabbit or anti-mouse fluorescent secondary antibodies (Abbkine, 1:200) at room temperature for 1 h, all sections were stained with DAPI (LEAGENE; DA0001) and observed under the fluorescence microscope.

Techniques: Activation Assay, Western Blot, Expressing, Immunohistochemistry, Imaging

Journal: Chinese Medicine

Article Title: Moxibustion ameliorates abnormal subchondral bone remodeling by promoting ACSL1-mediated autophagy to degrade NLRP3 in osteoarthritis

doi: 10.1186/s13020-025-01182-2

Figure Lengend Snippet: Moxibustion-induced autophagy and lysosome enhancement suppresses NLRP3 inflammasome activation. A Schematic of moxibustion promoting autophagy-lysosome pathway for NLRP3 degradation. B Representative images of autophagosomes (blue), lysosomes (yellow), and autolysosomes (red) in subchondral bone cells by TEM. Scale bar: 0.5 μm. C , I Analysis of LAMP1 immunofluorescence staining (n = 5 per group). Scale bar: 50 μm; 10 μm. D , E Co-localization images of SQSTM1 and NLRP3 in subchondral bone (n = 5 per group). Scale bar: 50 μm; 10 μm. F – H Intensity of SQSTM1 and NLRP3 co-localization in different groups. J Western blot detection of LAMP1, SQSTM1, LC3II/LC3II, ATG5 and Beclin1 proteins. Data presented as the mean ± SD. ns, not significant, * p < 0.05, ** p < 0.01, *** p < 0.001

Article Snippet: Primary antibody against NLRP3 (Proteintech, 68102-1-I; 1:80), Caspase-1 (Abmart, M025280F; 1:150), SQSTM1 (Abmart, T55546F; 1:200), LAMP1 (Proteintech, 21997-1-AP; 1:200), LC3B (Proteintech, 18725-1-AP; 1:200), ATG5 (Abmart, T55766F; 1:100) and PPARγ (ZENBIO, 340844; 1:200) were added to the slices at 4 °C for over 12 h. After being incubated with anti-rabbit or anti-mouse fluorescent secondary antibodies (Abbkine, 1:200) at room temperature for 1 h, all sections were stained with DAPI (LEAGENE; DA0001) and observed under the fluorescence microscope.

Techniques: Activation Assay, Immunofluorescence, Staining, Western Blot

Journal: Chinese Medicine

Article Title: Moxibustion ameliorates abnormal subchondral bone remodeling by promoting ACSL1-mediated autophagy to degrade NLRP3 in osteoarthritis

doi: 10.1186/s13020-025-01182-2

Figure Lengend Snippet: Schematic of the improvement of moxibustion against abnormal subchondral bone remodeling in OA. Abnormal stimulation triggers the activation of the NLRP3 inflammasome, leading to subchondral bone deterioration in OA. After moxibustion treatment, ACSL1 expression is upregulated, promoting increased autophagy levels to suppress NLRP3 activation, thereby balancing osteoclastic and osteogenic activities in OA. Ultimately, moxibustion treatment ameliorates abnormal subchondral bone remodeling and delays OA progression

Article Snippet: Primary antibody against NLRP3 (Proteintech, 68102-1-I; 1:80), Caspase-1 (Abmart, M025280F; 1:150), SQSTM1 (Abmart, T55546F; 1:200), LAMP1 (Proteintech, 21997-1-AP; 1:200), LC3B (Proteintech, 18725-1-AP; 1:200), ATG5 (Abmart, T55766F; 1:100) and PPARγ (ZENBIO, 340844; 1:200) were added to the slices at 4 °C for over 12 h. After being incubated with anti-rabbit or anti-mouse fluorescent secondary antibodies (Abbkine, 1:200) at room temperature for 1 h, all sections were stained with DAPI (LEAGENE; DA0001) and observed under the fluorescence microscope.

Techniques: Activation Assay, Expressing