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: Screening and validation of potential compounds for IVDD. (A) An intersection analysis of four compound libraries (Neuronal Signaling Library, FDA-approved Library, Natural Products Library, and Angiogenesis-Related Library) identified five potential therapeutic candidates. (B) Schematic of the in vitro experimental design. (C) Heatmap displaying the relative mRNA expression levels of key genes, including Acan , Mmp3 , and Vegfa , in NPCs following treatment with the five candidate compounds (n = 3). (D) RT-qPCR analysis of the relative mRNA expression of neurovascular modulation genes ( Slit2 , Sema3a , and Netrin-1 ) in NPCs treated with H 2 O 2 with or without RSV (n = 3). Data represent 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: Biomarker Discovery, In Vitro, Expressing, Quantitative RT-PCR

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 attenuates IVDD by restoring matrix homeostasis in LSI and AFP mouse models. (A) The mice experimental design diagram. (B) Representative H&E and SOFG images, and IF staining for ACAN and MMP3 expression (n = 5). Scale bar, 100 μm. (C) Histological scoring of disc degeneration in LSI and sham mice (n = 5). (D–E) Quantitative analysis of ACAN and MMP3 fluorescence intensity (n = 5). (F) Representative H&E and SOFG staining images of disc sections (n = 5). Scale bar, 100 μm. (G) Histological scores of AFP disc sections in each treatment group (n = 5). (H–I) Quantification of ACAN and MMP3 fluorescence intensity in the AFP model (n = 5). 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, Fluorescence

Journal: ACS Nano

Article Title: Development of Intervertebral Disc Organoids through Directed Differentiation of Mesenchymal Stem Cells and Hierarchical 3D Printing

doi: 10.1021/acsnano.5c14391

Figure Lengend Snippet: Hierarchical 3D-printed IVDOs recapitulate the NP and AF structure. (a) Histological images of H&E, Masson, and safranin O/fast green staining. (b) Immunofluorescence staining results for COL II, COL I, and ACAN demonstrate that the HG and SilMA hydrogels and the NP and AF differentiation media had synergistic effects on the zone-specific matrix secretion of IVD organoids at 28 days. (c) Semiquantitative fluorescence intensity of COL2, COL1 and ACAN. * p < 0.05, ** p < 0.01, two-tailed unpaired t test. n = 3 independent experiments per group.

Article Snippet: Sections were incubated for 1 h at room temperature using COL-1 (Proteintech, 14695-1-AP), COL-2 (Proteintech, 28459–1-AP), and ACAN (Proteintech, 13880-1-AP) as primary antibodies.

Techniques: Staining, Immunofluorescence, Fluorescence, Two Tailed Test

Journal: Cellular & Molecular Biology Letters

Article Title: Dichotomous SMAD2/3 regulation and selective antihypertrophic activity of heparin during in vitro chondrogenesis of mesenchymal stromal cells

doi: 10.1186/s11658-026-00899-8

Figure Lengend Snippet: MSC in vitro chondrogenesis in heparin–PEG hydrogels versus pellet controls. MSCs were cultured as pellets (pellet control [ctrl]; 5 × 10 5 cells) or in heparin–PEG hydrogels (H–PEG hydrogel; 1.2 × 10 6 cells) containing 22.4 mg/mL crosslinked heparin and 120 ng TGF-β1 for 4 weeks in TGF-β1-containing (10 ng/mL) standard chondrogenic medium in vitro. A , B Microsections of day-28 samples were assessed either via immunohistochemistry to detect type II collagen or aggrecan, and type X collagen, or by enzymatic activity staining to visualize alkaline phosphatase, as indicated (scale bar: overview = 200 µm, magnification = 50 µm; positive and negative staining controls are shown in Supplementary Fig. S2). One representative result out of n = 5 experiments with independent MSC donor populations is shown. C Day-28 gene expression levels of chondrocyte and D hypertrophy markers, with CPSF6 and HPRT used as reference genes. The hypertrophy markers in day-28 samples are expressed as ratio to COL2A1 . n = 4 experiments using independent MSC donor populations. Box plots were built as described in the statistics section. * p \documentclass[12pt]{minimal} \usepackage{amsmath} \usepackage{wasysym} \usepackage{amsfonts} \usepackage{amssymb} \usepackage{amsbsy} \usepackage{mathrsfs} \usepackage{upgreek} \setlength{\oddsidemargin}{-69pt} \begin{document}$$\le $$\end{document} ≤ 0.05, Mann–Whitney U test

Article Snippet: Human type II collagens, type X collagens, or aggrecans were visualized after incubation with a mouse anti-type II collagen antibody (1:1000, same as for western blotting), a mouse anti-type X collagen antibody (1:1, as above), and a mouse anti-aggrecan antibody (1:25, clone HAG7D4 (7D4), no. SM1353, Acris/OriGene, Rockville, MD, USA), respectively, and an ALP-coupled goat anti-mouse immunoglobulin G secondary antibody (ImmunoLogic, WellMed, MS Arnhem, the Netherlands) using the ImmPACT ® Vector ® Red Substrate kit (Vector Laboratories, Newark, CA, USA).

Techniques: In Vitro, Cell Culture, Control, Immunohistochemistry, Activity Assay, Staining, Negative Staining, Gene Expression, MANN-WHITNEY

Journal: Cellular & Molecular Biology Letters

Article Title: Dichotomous SMAD2/3 regulation and selective antihypertrophic activity of heparin during in vitro chondrogenesis of mesenchymal stromal cells

doi: 10.1186/s11658-026-00899-8

Figure Lengend Snippet: Effect of soluble heparin on MSC in vitro chondrogenesis. MSCs were cultured as pellets for 28 days in standard chondrogenic medium (including 10 ng/mL TGF-β1 and 6.25 µg/mL insulin) supplemented with 0, 10, 100, or 700 µg/mL soluble heparin. A Paraffin microsections of day-28 samples were assessed for type II collagen and aggrecan via immunohistochemistry (scale bar: overview = 200 µm, magnification = 50 µm). Representative pictures of n = 5 experiments with independent MSCs are shown. B Type II collagen content per pellet was measured by ELISA ( n = 4). C Proteoglycan content per pellet was assessed using DMMB assay and normalized to DNA amounts per pellet ( n = 5). D Cell culture supernatants were collected from seven to nine pellets at weekly intervals and analyzed for PGE2 levels using ELISA. Box plots were built as described in the statistics section and analyzed using Mann–Whitney U test. No statistically significant differences were found between the groups ( p \documentclass[12pt]{minimal} \usepackage{amsmath} \usepackage{wasysym} \usepackage{amsfonts} \usepackage{amssymb} \usepackage{amsbsy} \usepackage{mathrsfs} \usepackage{upgreek} \setlength{\oddsidemargin}{-69pt} \begin{document}$$>$$\end{document} > 0.05). Line graphs show data as mean \documentclass[12pt]{minimal} \usepackage{amsmath} \usepackage{wasysym} \usepackage{amsfonts} \usepackage{amssymb} \usepackage{amsbsy} \usepackage{mathrsfs} \usepackage{upgreek} \setlength{\oddsidemargin}{-69pt} \begin{document}$$\pm $$\end{document} ± standard error of the mean (SEM). # p \documentclass[12pt]{minimal} \usepackage{amsmath} \usepackage{wasysym} \usepackage{amsfonts} \usepackage{amssymb} \usepackage{amsbsy} \usepackage{mathrsfs} \usepackage{upgreek} \setlength{\oddsidemargin}{-69pt} \begin{document}$$\le $$\end{document} ≤ 0.05 versus control, paired Student’s t -test

Article Snippet: Human type II collagens, type X collagens, or aggrecans were visualized after incubation with a mouse anti-type II collagen antibody (1:1000, same as for western blotting), a mouse anti-type X collagen antibody (1:1, as above), and a mouse anti-aggrecan antibody (1:25, clone HAG7D4 (7D4), no. SM1353, Acris/OriGene, Rockville, MD, USA), respectively, and an ALP-coupled goat anti-mouse immunoglobulin G secondary antibody (ImmunoLogic, WellMed, MS Arnhem, the Netherlands) using the ImmPACT ® Vector ® Red Substrate kit (Vector Laboratories, Newark, CA, USA).

Techniques: In Vitro, Cell Culture, Immunohistochemistry, Enzyme-linked Immunosorbent Assay, Dimethylmethylene Blue Assay, MANN-WHITNEY, Control