Journal: Materials Today Bio

Article Title: Modulating cell surface chemistry through mild reduction reinforces extracellular-to-intracellular transmission forces and mechano-signaling

doi: 10.1016/j.mtbio.2026.102908

Figure Lengend Snippet: Mild reduction of the cell surface promotes cell–matrix adhesion and focal adhesion assembly on adipose-like stiffness hydrogels. (A) Schematic representation of cell culture on collagen type 1-conjugated polyacrylamide hydrogels with adipose tissue-like stiffness. This schematic was created using BioRender.com . (B) Representative immunofluorescence staining for F-actin (red) and pFAK (green). (C-H) Quantitative image analysis of cell morphology and focal adhesion (FA) parameters. scale bar = 50 μm, Data are presented as mean ± 95% confidence interval (CI). ∗∗ p < 0.01; ∗∗∗ p < 0.001.

Article Snippet: Briefly, cells were seeded onto collagen type 1-conjugated polyacrylamide hydrogels and cultured in DMEM supplemented with 10% v/v FBS (26140-079, Gibco, Waltham, MA, USA).

Techniques: Cell Culture, Immunofluorescence, Staining

Journal: Bioactive Materials

Article Title: Bioinspired scaffold recapitulating chondrogenic ontogeny and microenvironment for functional cartilage regeneration

doi: 10.1016/j.bioactmat.2025.11.041

Figure Lengend Snippet: Biological functional analysis of tri-layer scaffold systems. A) Representative images of migrated cells stained with crystal violet at 48 h. B) Number of migrating cells in control and CE-SKP. C) Glycosaminoglycan concentrations in control and CE-SKP at 7 and 14 days. D) qPCR analysis for COL2, SOX9, and ACAN. E) Western blotting of COL2 and SOX9. F) Alizarin red staining of control and CPH. G) ALP activity of control and CPH. H) qPCR analysis for COL1, COL10, OCN, and RUNX2. I) Western blotting of COL10, COL1, and RUNX2. J) Representative images of migrated cells stained with crystal violet at 48 h. K) Number of migrating cells in control and P2G3. L) Scanning electron microscope micrographs of P2G3. ∗P < 0.05, ∗∗P < 0.01, ∗∗∗P < 0.001.

Article Snippet: After blocking the membranes for an hour at room temperature using fast blocking solution, the membranes were incubated with primary antibodies specific for SOX9 (1:2000, 67439-1-Ig, Proteintech), COL2 (1:200, NBP1-91056, Novus Biologicals), COL10 (1:500, 26984-1-AP, Proteintech), COL1 (1:2000, 67288-1-Ig, Proteintech), RUNX2 (1:200, 20700-1-AP, Proteintech), FGF2 (1:500, 11234-1-AP, Proteintech), FGF18 (1:500, 60341-1-Ig, Proteintech) and GAPDH (1:50000, 60004-1-Ig, Proteintech) for an entire night at 4 °C.

Techniques: Functional Assay, Staining, Control, Western Blot, Activity Assay, Microscopy

Journal: Bioactive Materials

Article Title: Bioinspired scaffold recapitulating chondrogenic ontogeny and microenvironment for functional cartilage regeneration

doi: 10.1016/j.bioactmat.2025.11.041

Figure Lengend Snippet: Regeneration of the femoral trochlea in a rabbit model. Representative images of IHC staining for A) COL1, B) COL10.

Article Snippet: After blocking the membranes for an hour at room temperature using fast blocking solution, the membranes were incubated with primary antibodies specific for SOX9 (1:2000, 67439-1-Ig, Proteintech), COL2 (1:200, NBP1-91056, Novus Biologicals), COL10 (1:500, 26984-1-AP, Proteintech), COL1 (1:2000, 67288-1-Ig, Proteintech), RUNX2 (1:200, 20700-1-AP, Proteintech), FGF2 (1:500, 11234-1-AP, Proteintech), FGF18 (1:500, 60341-1-Ig, Proteintech) and GAPDH (1:50000, 60004-1-Ig, Proteintech) for an entire night at 4 °C.

Techniques: Immunohistochemistry

Journal: Bioactive Materials

Article Title: An intelligent nanoliposome alleviates disc degeneration and discogenic pain by inhibiting neurovascular ingrowth via a “Soil-conditioning, seed-modulating, and weeds-suppressing” strategy

doi: 10.1016/j.bioactmat.2025.11.048

Figure Lengend Snippet: Oxidative stress and aberrant neurovascularization are progressively enhanced during IVDD. (A) Representative T2-weighted MRI images showing disc structure in human intervertebral discs at varying Pfirrmann degeneration grades (II–IV). (B–C) Fluorescent staining and quantitative analysis of ROS levels in primary NPCs from grades II–IV NP tissues, showing a significant increase in oxidative stress with disease severity (n = 5). Scale bar, 20 μm. (D–F) IHC staining and quantitative analyses of CD31 and Tuj1 across different degenerative grades (n = 5). Scale bar, 50 μm. (G) H&E, SOFG, and IF staining of COL2A1 and MMP3 for IVDs (n = 5). Scale bar, 100 μm. (H) Histological score of the IVD tissues (n = 5). (I–J) Quantification of IF staining for COL2A1 and MMP3 (n = 5). (K–M) IF staining and corresponding quantification analyses of CD31 and Tuj1 in disc tissues of sham and AFP-treated mice (n = 5). Scale bar, 100 μm. Data are presented as mean ± SD. The comparisons among groups were performed using one-way ANOVA. ∗, P < 0.05; ∗∗, P < 0.01; ∗∗∗, P < 0.001; ∗∗∗∗, P < 0.0001; ns = not significant.

Article Snippet: For immunofluorescence (IF) staining, antigen retrieval was conducted using citrate buffer (0.1 M, pH 6.0), followed by blocking in 10 % normal goat serum (Solarbio, Cat. No. SL038) at room temperature for 1 h. Sections were incubated overnight at 4 °C with primary antibodies targeting ACAN (Cat. No. DF7561, Affinity), COL2A1 (Cat. No. AF0135, Affinity), MMP3 (Cat. No. AF0217, Affinity), MMP13 (Cat. No. 18165-1-AP, Proteintech), CD31 (Cat. No. AF0077, Affinity), p-LATS1 (Cat. No. AF7170, Affinity), p-YAP (Cat. No. AF3328, Affinity), Tuj1 (Cat. No. 66375, Proteintech), CGRP (Cat. No. DF7386, Affinity), SP (Cat. No. DF7522, Affinity), and PGP9.5 (Cat. No. AF5490, Affinity) followed by incubation with appropriate fluorescent secondary antibodies for 2 h at room temperature and counterstaining with DAPI (Beyotime, Cat. No. C1006).

Techniques: Staining, Immunohistochemistry

Journal: Bioactive Materials

Article Title: An intelligent nanoliposome alleviates disc degeneration and discogenic pain by inhibiting neurovascular ingrowth via a “Soil-conditioning, seed-modulating, and weeds-suppressing” strategy

doi: 10.1016/j.bioactmat.2025.11.048

Figure Lengend Snippet: NM-LP TK /RSV-MnCDs attenuates H 2 O 2 -induced ECM degradation by enhancing anabolic gene expression and suppressing catabolic activity in NPCs. (A–D) RT-qPCR analysis of mRNA expression levels of anabolic markers Acan and Col2a1 , and catabolic enzymes Adamts5 and Mmp3 in NPCs (n = 3). (E–G) Representative IF images and quantitative analyses of ACAN and MMP3 expression in NPCs across treatment groups, showing reduced ECM degradation in NM-LPTK/RSV-MnCDs-treated NPCs (n = 3). Scale bar, 30 μm. Data are presented as mean ± SD. The comparisons among groups were performed using one-way ANOVA. ∗, P < 0.05; ∗∗, P < 0.01; ∗∗∗, P < 0.001; ∗∗∗∗, P < 0.0001; ns = not significant.

Article Snippet: For immunofluorescence (IF) staining, antigen retrieval was conducted using citrate buffer (0.1 M, pH 6.0), followed by blocking in 10 % normal goat serum (Solarbio, Cat. No. SL038) at room temperature for 1 h. Sections were incubated overnight at 4 °C with primary antibodies targeting ACAN (Cat. No. DF7561, Affinity), COL2A1 (Cat. No. AF0135, Affinity), MMP3 (Cat. No. AF0217, Affinity), MMP13 (Cat. No. 18165-1-AP, Proteintech), CD31 (Cat. No. AF0077, Affinity), p-LATS1 (Cat. No. AF7170, Affinity), p-YAP (Cat. No. AF3328, Affinity), Tuj1 (Cat. No. 66375, Proteintech), CGRP (Cat. No. DF7386, Affinity), SP (Cat. No. DF7522, Affinity), and PGP9.5 (Cat. No. AF5490, Affinity) followed by incubation with appropriate fluorescent secondary antibodies for 2 h at room temperature and counterstaining with DAPI (Beyotime, Cat. No. C1006).

Techniques: Gene Expression, Activity Assay, Quantitative RT-PCR, Expressing

Journal: Bioactive Materials

Article Title: An intelligent nanoliposome alleviates disc degeneration and discogenic pain by inhibiting neurovascular ingrowth via a “Soil-conditioning, seed-modulating, and weeds-suppressing” strategy

doi: 10.1016/j.bioactmat.2025.11.048

Figure Lengend Snippet: NM-LP TK /RSV-MnCDs suppresses vascular ingrowth and matrix degradation in AFP-induced disc degeneration through Hippo pathway inactivation. (A–D) Representative IF images and quantitative analyses of CD31 and Tuj1 in IVDs from AFP-induced IVDD mice treated with MCB, TRULI, or their combinations with NM-LP TK /RSV-MnCDs (n = 5). Scale bar, 100 μm. (E–H) IF staining and quantification of COL2A1 and MMP13 expression in NP regions (n = 5). Scale bar, 100 μm. Data are presented as mean ± SD. The comparisons among groups were performed using one-way ANOVA. ∗, P < 0.05; ∗∗, P < 0.01; ∗∗∗, P < 0.001; ∗∗∗∗, P < 0.0001; ns = not significant.

Article Snippet: For immunofluorescence (IF) staining, antigen retrieval was conducted using citrate buffer (0.1 M, pH 6.0), followed by blocking in 10 % normal goat serum (Solarbio, Cat. No. SL038) at room temperature for 1 h. Sections were incubated overnight at 4 °C with primary antibodies targeting ACAN (Cat. No. DF7561, Affinity), COL2A1 (Cat. No. AF0135, Affinity), MMP3 (Cat. No. AF0217, Affinity), MMP13 (Cat. No. 18165-1-AP, Proteintech), CD31 (Cat. No. AF0077, Affinity), p-LATS1 (Cat. No. AF7170, Affinity), p-YAP (Cat. No. AF3328, Affinity), Tuj1 (Cat. No. 66375, Proteintech), CGRP (Cat. No. DF7386, Affinity), SP (Cat. No. DF7522, Affinity), and PGP9.5 (Cat. No. AF5490, Affinity) followed by incubation with appropriate fluorescent secondary antibodies for 2 h at room temperature and counterstaining with DAPI (Beyotime, Cat. No. C1006).

Techniques: Staining, Expressing

Journal: International Journal of Molecular Medicine

Article Title: Deciphering the CAF-LCN2 axis: Key to overcoming anti-PD-L1 immunotherapy resistance in lung cancer

doi: 10.3892/ijmm.2026.5735

Figure Lengend Snippet: Influence of LCN2 on the growth of lung cancer in mice. (A) Simplified flowchart outlining the establishment of an anti-PD-L1 immunotherapy tolerance model in mice with lung cancer. (B) Western blot analysis of LCN2, COL1A1 and THY1 expression in tumor tissues from drug-resistant and drug-sensitive mice. (C) Immunohistochemical detection of LCN2, COL1A1 and THY1 expression in tumor tissues from drug-resistant and drug-sensitive mice (scale bar, 100 µm). (D) Establishment of a model for anti-PD-L1 immunotherapy in LCN2-silenced mice. (E) Evaluation of LCN2 mRNA expression in tumor tissues after LCN2 gene intervention in mice. (F) Assessment of LCN2 protein expression in tumor tissues following LCN2 gene intervention in mice. (G) Tumor size in LCN2 gene intervention mice (n=10 in each group). *P<0.05, **P<0.01 and ***P<0.001. Unpaired two-tailed Student's t-test was used for (B) and (C) and one-way ANOVA with Tukey's multiple comparisons test was used for (E) and (G). LCN2, lipocalin 2; PD-L1, programmed death-ligand 1; sh-, short hairpin; NC, negative control; COL1A1, collagen α-1(I) chain; THY1, Thy-1 membrane glycoprotein; i.p., intraperitoneal.

Article Snippet: Following permeabilization, cells were incubated overnight at 4°C with the following primary antibodies: Rabbit anti-collagen α-1(I) chain (COL1A1) (cat. no. PA5-101300; 1:250; Invitrogen; Thermo Fisher Scientific, Inc.), rabbit anti-Cytokeratin (cat. no. ab53280; 1:500; Abcam) and mouse anti-CD31 (cat. no. sc-376764; 1:50; Santa Cruz Biotechnology, Inc.).

Techniques: Western Blot, Expressing, Immunohistochemical staining, Two Tailed Test, Negative Control, Membrane