aggrecan  (R&D Systems)

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: Non-coding RNA Research

Article Title: Hsa_circ_0101645 contributes to excessive autophagy and apoptosis in intervertebral disc degeneration by acting as a miR-1304-5p sponge modulating BNIP3 expression

doi: 10.1016/j.ncrna.2025.11.007

Figure Lengend Snippet: Hsa_circ_0101645 accelerating the IVDD process in vivo . A: Diagram of the animal procedure for this study. B: Grouping information for this section. C: Representative X-rays of each group of rats. Statistical graph demonstrating the disc height index (DHI) changes for L4/5 in each group of rats (N = 6) (One-way ANOVA test with Tukey's multiple comparisons test). D: HE staining exhibiting pathological changes of CEP, NP, and AP in IVD in each group of rats (N = 6). Scale bar: 500 μm. E: EdU staining was used to detect cell proliferation in IVD tissues of rats (N = 6) (One-way ANOVA test with Tukey's multiple comparisons test). Scale bar: 50 μm. F: TUNEL (white light) staining exhibiting TUNEL-positive cells in IVD tissues of rats (N = 6) (One-way ANOVA test with Tukey's multiple comparisons test). Scale bar: 50 μm. G: The effect of hsa_circ_0101645 on the protein levels of Collagen Ⅱ, Aggrecan, MMP-3 and MMP-13 in IVD was observed by IHC staining (N = 6) (One-way ANOVA test with Tukey's multiple comparisons test). Scale bar: 100 μm. H-I: The expression of hsa_circ_0101645 (H) and miR-1304-5p (I) in each group of IVD tissues (N = 6) (One-way ANOVA test with Tukey's multiple comparisons test). J-K: Changes in expression of apoptosis ( J; Caspase 3, Bcl-2 and Bax) and autophagy markers ( K; LC3B, Beclin and P62) in IVD tissues (N = 3) (One-way ANOVA test with Tukey's multiple comparisons test or Kruskal-Wallis test with Dunn's multiple comparisons test). ∗ indicates P < 0.05.

Article Snippet: Sections were then incubated with primary antibodies against Collagen II (28459-1-AP; 1:200; Proteintech, USA), Aggrecan (13880-1-AP; 1:100; Proteintech), MMP-3 (17873-1-AP; 1:200; Proteintech), and MMP-13 (18165-1-AP; 1:100; Proteintech), followed by the corresponding secondary antibodies.

Techniques: In Vivo, Staining, TUNEL Assay, Immunohistochemistry, 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: 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