Journal: Bioactive Materials

Article Title: Dual-function thermoresponsive antibiotic-loaded hydrogel with antimicrobial and osteogenic properties for implant-related infection control

doi: 10.1016/j.bioactmat.2026.02.044

Figure Lengend Snippet: In vivo immunohistochemical analysis (n = 6). (A) Representative immunolabeling images of the Control, PNVCL, and PNVCL/TC 25 mg/mL groups showing osteopontin (OPN) and osteocalcin (OCN) expression (immunopositive cells indicated by black arrows). (B) Mean scores (0–3) ± standard deviation for OPN immunostaining. (C) Mean scores (0–3) ± standard deviation for OCN immunostaining. Bars indicate statistically significant differences between groups (p < 0.05; Tukey HSD test).

Article Snippet: Endogenous peroxidase activity was quenched by incubation with 3% hydrogen peroxide for 1 h, followed by blocking of nonspecific binding sites with 1% bovine serum albumin for 12 h. The sections were then incubated with goat anti-osteopontin and goat anti-osteocalcin primary antibodies (sc-21742 and sc-30044, respectively; Santa Cruz Biotechnology, Dallas, TX, USA).

Techniques: In Vivo, Immunohistochemical staining, Immunolabeling, Control, Expressing, Standard Deviation, Immunostaining

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: Bioactive Materials

Article Title: Precisely regulated physically-crosslinked carriers enable synergetic release of bioactive factors for MSC-mediated cartilage regeneration

doi: 10.1016/j.bioactmat.2026.01.009

Figure Lengend Snippet: Biocompatibility and Bioactivity of PSF and KSF in vivo . (a) Transwell Assay of MSCs after treated with PBS, MAP and PSF. Scale bar = 200 μm. (b) Wound Healing Assay of MSCs at 0h and 24h. Scale bar = 200 μm. (c) Immunofluorescent staining of cell pellets after 21 days of co-culture. Blue: DAPI; Green: ActinGreen; Red: Aggrecan. Scale bar = 200 μm. (d) Alcian blue staining of 2D cultured MSCs after 14 days. Scale bar = 200 μm. (e) Cell viability of MSCs at day 3 after co-culture. (f) The cell number of MSCs migrated from upper to lower chamber in Transwell assay. (g) The average distance MSCs migrated from injured margin in wound healing assay. (h–j) qRT-PCR of Col2a1 , Acan and Sox9 mRNA relative expression ratio compared with PBS group. ns: p > 0.05; ∗ p < 0.05; ∗∗ p < 0.01; ∗∗∗ p < 0.001; ∗∗∗∗ p < 0.0001.

Article Snippet: Immunofluorescent staining was performed with Aggrecan primary antibody (13880-1-AP, Proteintech, USA), ActinGreen ( R37110 , Thermo, USA) and DAPI (Solarbio, China) and observed with 3D reconstruction under high content imaging system (PerkinElmer, Operetta CLS, USA).

Techniques: In Vivo, Transwell Assay, Wound Healing Assay, Staining, Co-Culture Assay, Cell Culture, Quantitative RT-PCR, Expressing

Journal: The Journal of General Physiology

Article Title: Beat-locked ATP microdomains in the sinoatrial node map a Ca 2+ -timed energetic hierarchy and regional pacemaker roles

doi: 10.1085/jgp.202513874

Figure Lengend Snippet: Superior SA node myocytes exhibit elevated diastolic ATP and metabolic flux compared with the inferior region. (A) 3D segmented maximum-intensity projection of a whole-mount SA node immunolabeled for CD31 (vasculature, red) and cyto-iATP (myocytes, green). The dashed line denotes the boundary between superior and inferior regions. (B) Image-processing workflow illustrating merged maximum-intensity projections, binary segmentation masks, and extraction of grayscale cyto-iATP signals used for quantitative analysis. (C) Mean cyto-iATP fluorescence intensity per myocyte, grouped by region ( N = 5 mice per region), reporting expression levels of the EGFP-tagged cyto-iATP sensor. (D) Live confocal imaging of cyto-iATP signals showing representative line-scan images and corresponding normalized fluorescence traces (F/F 0 ) from superior and inferior regions. (E and F) Summary quantification of cyto-iATP signal mass rate (E) and estimated diastolic [ATP] i (F). P values are shown above comparisons. Large circles denote per-animal means; small circles indicate individual biological replicates. N represents the number of independent mice.

Article Snippet: For immunolabeling, SA nodes were incubated for 48 h at 4°C with a goat anti-mouse CD31 primary antibody (1:50, AF3628; R&D Systems).

Techniques: Immunolabeling, Extraction, Fluorescence, Expressing, Imaging

Journal: Bioactive Materials

Article Title: Ribbon-shaped microgels as bioinks for 3D bioprinting of anisotropic tissue structures

doi: 10.1016/j.bioactmat.2025.12.040

Figure Lengend Snippet: μRB bioinks modulate MSC morphology, osteogenesis and bone formation in a stiffness-dependent manner. (A) Live/dead staining of MSCs after extrusion (Day 0) or after 28 days of culture in osteogenic medium. Green: live cells, Red: dead cells. (B) Metabolic activity of MSCs measured by PrestoBlue assay at day 0 and day 14 after bioprinting (n = 6 per group). (C) DNA content per scaffold measured by PicoGreen assay at day 28 (n = 3 per group). (D) Alizarin red S (ARS) staining for mineralized bone matrix, (E) Aniline Blue staining for total collagen, and (F) immunostaining of Osteocalcin (OCN), a mature bone marker, at day 14 and day 21 of osteogenesis (n = 4 per group). (G–I) Quantification of ARS and Aniline Blue percent positive area and OCN mean fluorescence intensity (MFI). Scale bar = 100 μm. Values are presented as mean ± S.D. and p-values were determined by one-way analysis of variance (ANOVA) with Tukey's multiple comparisons test; ∗p ≤ 0.05, ∗∗p ≤ 0.01, ∗∗∗p ≤ 0.005, ∗∗∗∗p ≤ 0.001.

Article Snippet: Primary antibody for osteocalcin (1:200, 23418-1-AP, Proteintech), GFP (1:100, 50430-2-AP, Proteintech) was diluted in 1 % BSA with 0.1 % Triton X-100 and incubated overnight at 4 °C.

Techniques: Staining, Activity Assay, Prestoblue Assay, Picogreen Assay, Immunostaining, Marker, Fluorescence

Journal: Biomedical Reports

Article Title: ALDH5A1 as a key in osteonecrosis of the femoral head: Insights from bioinformatics and experimental validation

doi: 10.3892/br.2026.2136

Figure Lengend Snippet: Accuracy analysis of signature gene. (A) Differential violin plots and ROC curves for ALDH5A1 in experimental groups. (B) Differential violin plots and ROC curves for ALDH5A1 in validation groups. * P<0.05 and *** P<0.001. ROC, receiver operating characteristic; ALDH5A1, Aldehyde dehydrogenase 5 family member A1; AUC, area under curve.

Article Snippet: After washing, cells were incubated overnight at 4 ̊C with anti-ALDH5A1 primary antibody (Proteintech, China), followed by 2-h incubation with FITC-conjugated goat anti-rabbit IgG secondary antibody (Epizyme Biotech, China) in the dark.

Techniques: Biomarker Discovery

Journal: Biomedical Reports

Article Title: ALDH5A1 as a key in osteonecrosis of the femoral head: Insights from bioinformatics and experimental validation

doi: 10.3892/br.2026.2136

Figure Lengend Snippet: Signature gene correlation analysis. (A) Heatmap of ALDH5A1-related DEGs. (B) Volcano plot of ALDH5A1-related DEGs. (C) Bubble plot of ALDH5A1-related DEGs. ALDH5A1, Aldehyde dehydrogenase 5 family member A1; DEGs, differentially expressed genes.

Article Snippet: After washing, cells were incubated overnight at 4 ̊C with anti-ALDH5A1 primary antibody (Proteintech, China), followed by 2-h incubation with FITC-conjugated goat anti-rabbit IgG secondary antibody (Epizyme Biotech, China) in the dark.

Techniques:

Journal: Biomedical Reports

Article Title: ALDH5A1 as a key in osteonecrosis of the femoral head: Insights from bioinformatics and experimental validation

doi: 10.3892/br.2026.2136

Figure Lengend Snippet: GO and GSVA enrichment analysis. (A) Bar plot of GO function enrichment analysis. (B) Bubble plot of GO enrichment analysis (C) GSVA analysis based on ALDH5A1 expression levels. GO, Gene Ontology; GSVA, Gene Set Variation Analysis; DEGs, differentially expressed genes; ALDH5A1, Aldehyde dehydrogenase 5 family member A1; KEGG, Kyoto Encyclopedia of Genes and Genomes.

Article Snippet: After washing, cells were incubated overnight at 4 ̊C with anti-ALDH5A1 primary antibody (Proteintech, China), followed by 2-h incubation with FITC-conjugated goat anti-rabbit IgG secondary antibody (Epizyme Biotech, China) in the dark.

Techniques: Expressing

Journal: Biomedical Reports

Article Title: ALDH5A1 as a key in osteonecrosis of the femoral head: Insights from bioinformatics and experimental validation

doi: 10.3892/br.2026.2136

Figure Lengend Snippet: Gene set enrichment analysis. (A) Functional enrichment analysis of GSEA in the ALDH5A1 high expression group. (B) Functional enrichment analysis of GSEA in the ALDH5A1 low expression group. GSEA, Gene Set Enrichment Analysis; ALDH5A1, Aldehyde dehydrogenase 5 family member A1; KEGG, Kyoto Encyclopedia of Genes and Genomes.

Article Snippet: After washing, cells were incubated overnight at 4 ̊C with anti-ALDH5A1 primary antibody (Proteintech, China), followed by 2-h incubation with FITC-conjugated goat anti-rabbit IgG secondary antibody (Epizyme Biotech, China) in the dark.

Techniques: Functional Assay, Expressing

Journal: Biomedical Reports

Article Title: ALDH5A1 as a key in osteonecrosis of the femoral head: Insights from bioinformatics and experimental validation

doi: 10.3892/br.2026.2136

Figure Lengend Snippet: Differences in 29 immune-related functions across different ALDH5A1 expression levels. * P<0.05. ONFH, osteonecrosis of the femoral head; ssGSEA, single-sample Gene Set Enrichment Analysis; ALDH5A1, aldehyde dehydrogenase 5 family member A1.

Article Snippet: After washing, cells were incubated overnight at 4 ̊C with anti-ALDH5A1 primary antibody (Proteintech, China), followed by 2-h incubation with FITC-conjugated goat anti-rabbit IgG secondary antibody (Epizyme Biotech, China) in the dark.

Techniques: Expressing

Journal: Biomedical Reports

Article Title: ALDH5A1 as a key in osteonecrosis of the femoral head: Insights from bioinformatics and experimental validation

doi: 10.3892/br.2026.2136

Figure Lengend Snippet: Immune correlation analysis. (A) Lollipop plot of correlations between ALDH5A1 expression and immune cell infiltration. The X-axis shows the Spearman correlation coefficient. The size of the circles corresponds to the absolute correlation coefficient (|Correlation|), and the color indicates statistical significance (Red for P<0.05, Grey for P≥0.05). The P-values are displayed on the right. (B) Distribution of Spearman correlation of activated CD4 memory T cells with ALDH5A1 in samples from patients with ONFH. ALDH5A1, Aldehyde dehydrogenase 5 family member A1; ONFH, osteonecrosis of the femoral head.

Article Snippet: After washing, cells were incubated overnight at 4 ̊C with anti-ALDH5A1 primary antibody (Proteintech, China), followed by 2-h incubation with FITC-conjugated goat anti-rabbit IgG secondary antibody (Epizyme Biotech, China) in the dark.

Techniques: Expressing

Journal: Biomedical Reports

Article Title: ALDH5A1 as a key in osteonecrosis of the femoral head: Insights from bioinformatics and experimental validation

doi: 10.3892/br.2026.2136

Figure Lengend Snippet: Plot of the competing endogenous RNA network of ALDH5A1. ALDH5A1, Aldehyde dehydrogenase 5 family member A1.

Article Snippet: After washing, cells were incubated overnight at 4 ̊C with anti-ALDH5A1 primary antibody (Proteintech, China), followed by 2-h incubation with FITC-conjugated goat anti-rabbit IgG secondary antibody (Epizyme Biotech, China) in the dark.

Techniques:

Journal: Biomedical Reports

Article Title: ALDH5A1 as a key in osteonecrosis of the femoral head: Insights from bioinformatics and experimental validation

doi: 10.3892/br.2026.2136

Figure Lengend Snippet: Verification of ALDH5A1 expression. (A) RT-qPCR analysis of ALDH5A1 mRNA levels in chondrocytes from the NC and ONFH groups. (B) Densitometric quantification of ALDH5A1 protein expression in the NC and ONFH groups. (C) Representative western blot images showing ALDH5A1 and the loading control GAPDH in the NC and ONFH groups (n=3). (D) Immunofluorescence staining of ALDH5A1 in chondrocytes from the NC and ONFH groups. ALDH5A1 is shown in green and nuclei are counterstained with DAPI (blue). (E) Quantification of the relative fluorescence intensity of ALDH5A1 in the NC and ONFH groups. * P<0.05 and ** P<0.01. ALDH5A1, aldehyde dehydrogenase 5 family member A1; ONFH, osteonecrosis of the femoral head; NC, normal control.

Article Snippet: After washing, cells were incubated overnight at 4 ̊C with anti-ALDH5A1 primary antibody (Proteintech, China), followed by 2-h incubation with FITC-conjugated goat anti-rabbit IgG secondary antibody (Epizyme Biotech, China) in the dark.

Techniques: Expressing, Quantitative RT-PCR, Western Blot, Control, Immunofluorescence, Staining, Fluorescence

Journal: Redox Biology

Article Title: Aryl hydrocarbon receptor in club cells drives Th17-mediated lung injury following inhalation exposure to environmentally persistent free radicals

doi: 10.1016/j.redox.2026.104105

Figure Lengend Snippet: Generation and validation of club cell-specific AHR knockout mice ( Ahr ΔCC). (a) Schematic of breeding strategy to generate Ahr ΔCC mice by crossing Ahr fl/fl mice with Scgb1a1-CreER TM mice, followed by tamoxifen induction. (b) Immunofluorescence staining showing club cell marker CC10 (green), AHR (red) and DAPI (blue) in lung sections of Cre-negative Ahr fl/fl LM control (top panel) and Ahr ΔCC (bottom panel) mice. (c) Representative flow cytometry plots of lung epithelial cells gated as CD45 − CD31 − EpCAM + CC10 + cells isolated from lungs of LM (left) or Ahr ΔCC (center) mice and Fluorescence-minus-one (FMO) control for AHR staining (right). (d) Quantification of the percentage of AHR + CC10 + cells in the lungs of LM (white bar) and Ahr ΔCC (gray bar) mice. Data represent mean ± SEM, n = 3-4 mice per group. Statistical significance was determined using Student's t-test; ∗p < 0.05.

Article Snippet: After washing, slides were incubated overnight at room temperature with a second primary antibody against CC10 (Proteintech, Rosemont, IL; catalog# 10490-1-AP), followed by an Alexa Fluor Plus 488-conjugated goat anti-rabbit secondary antibody (Invitrogen, Waltham, MA; catalog# A32731).

Techniques: Biomarker Discovery, Knock-Out, Immunofluorescence, Staining, Marker, Control, Flow Cytometry, Isolation, Fluorescence