antibodies targeting acan  (Proteintech)

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: Regenerative Therapy

Article Title: Hypoxia-preconditioned bone marrow mesenchymal stem cell-derived exosomes ameliorate knee osteoarthritis by promoting cartilage regeneration and alleviating pain in rats

doi: 10.1016/j.reth.2025.101049

Figure Lengend Snippet: Hypo-BMSCs-Exos attenuate IL-1β-induced chondrocyte injury. A: Immunofluorescence staining reveals changes in Collagen II, Aggrecan, ADAMTS-5, and MMP-13 in IL-1β-induced chondrocytes following intervention with Norm-BMSCs-Exos and Hypo-BMSCs-Exos. B: Quantitative analysis of immunofluorescence staining. C: Western blot detection of protein expression levels of Collagen II, Aggrecan, ADAMTS-5, and MMP-13 in IL-1β-induced chondrocytes treated with Norm-BMSCs-Exos and Hypo-BMSCs-Exos. D: Statistical analysis of relative protein band intensity values. Data are presented as Mean ± SD. Intergroup differences with statistical significance: ∗P < 0.05, ∗∗P < 0.01, ∗∗∗P < 0.001, n = 3 per group.

Article Snippet: The cells were then washed three times with PBS, with each wash lasting 5 min. After blocking with bovine serum albumin (BSA) for 20 min at room temperature and subsequent washing with PBS, the cells were incubated overnight at 4 °C with primary antibodies against iNOS (Servicebio, China), COX2 (Servicebio, China), Collagen II (Bisso, China), aggrecan (Bisso, China), ADAMTS-5 (Bisso, China), MMP-13 (Affinity, USA), p16 (Abcam, USA), p21 (Affinity, USA), and p53 (Proteintech, USA).

Techniques: Immunofluorescence, Staining, Western Blot, Expressing

Journal: Regenerative Therapy

Article Title: Hypoxia-preconditioned bone marrow mesenchymal stem cell-derived exosomes ameliorate knee osteoarthritis by promoting cartilage regeneration and alleviating pain in rats

doi: 10.1016/j.reth.2025.101049

Figure Lengend Snippet: Intra-articular injection of Hypo-BMSCs-Exo protects articular cartilage from degeneration in KOA rats. A: Representative images of immunohistochemical staining for iNOS, COX2, Collagen II, Aggrecan, ADAMTS-5, and MMP-13 in cartilage tissue at 8 weeks post-intervention. Scale bar = 50 μm. B: Quantitative analysis and statistical evaluation of immunopositive cells. Data are expressed as mean ± standard deviation (SD), with n = 3 per group. Intergroup statistical significance is indicated as: ∗P < 0.05, ∗∗P < 0.01, ∗∗∗P < 0.001 compared to control rats; # p < 0.05、 ## p < 0.01、 ### p < 0.001 indicate differences compared with OA rats; & p < 0.05、 && p < 0.01、 &&& p < 0.001 indicate differences compared with Norm-BMSCs-Exos rats.

Article Snippet: The cells were then washed three times with PBS, with each wash lasting 5 min. After blocking with bovine serum albumin (BSA) for 20 min at room temperature and subsequent washing with PBS, the cells were incubated overnight at 4 °C with primary antibodies against iNOS (Servicebio, China), COX2 (Servicebio, China), Collagen II (Bisso, China), aggrecan (Bisso, China), ADAMTS-5 (Bisso, China), MMP-13 (Affinity, USA), p16 (Abcam, USA), p21 (Affinity, USA), and p53 (Proteintech, USA).

Techniques: Injection, Immunohistochemical staining, Staining, Standard Deviation, Control

Journal: Aging and Disease

Article Title: Enhanced Chondrogenic Potential and Osteoarthritis Treatment Using Cyaonoside A-Induced MSC Delivered via a Hyaluronic Acid-Based Hydrogel System

doi: 10.14336/AD.2024.10016

Figure Lengend Snippet: CyA promotes chondrogenic differentiation of MSC . ( A ) Relative mRNA expression levels of ACAN and COL2A in MSC. ( B ) Alcian blue staining images of MSC cultured with CyA for 21 days. ( C ) Cell viability of MSC over 21 days. ( D ) Relative mRNA expression levels of ACAN, COL2A, and SOX9 in MSC over 21 days. ( E ) Protein expression levels of ACAN, COL2A, and SOX9 in MSC over 21 days. ( F ) Representative immunocytochemistry images of COL2A in MSC from Day 0 to Day 21. Statistical significance was determined using one-way ANOVA for multiple group comparisons, followed by post hoc tests. All data are presented as mean ± SD (n = 3). *p < 0.05, **p < 0.01, ***p < 0.001, and ns = no statistically significant difference between groups.

Article Snippet: Overnight incubation at 4°C with diluted primary antibodies followed: ACAN (1:2000, Cat# 68350-1-Ig, Proteintech, USA), COL2A (1:2000, Cat# 28459-1-AP, Proteintech, USA), SOX9 (1:2000, Cat# 67439-1-Ig, Proteintech, USA), MMP13 (1:2000, Cat# 18165-1-AP, Proteintech, USA), CD9 (1:1000, Cat# 13174, CST, USA), CD63 (1:1000, Cat# 52090, CST, USA), GM130 (1:1000, Cat# 12480, CST, USA), Bcl-2 (1:1000, Cat# 3498, CST, USA), P53 (1:1000, Cat# 9282, CST, USA), and GAPDH (1:10000, Cat# 10494-1-AP, Proteintech, USA).

Techniques: Expressing, Staining, Cell Culture, Immunocytochemistry

Journal: Aging and Disease

Article Title: Enhanced Chondrogenic Potential and Osteoarthritis Treatment Using Cyaonoside A-Induced MSC Delivered via a Hyaluronic Acid-Based Hydrogel System

doi: 10.14336/AD.2024.10016

Figure Lengend Snippet: Expression of ACAN, COL2A, SOX9, and MMP13 in chondrocytes . ( A ) Western blot analysis of ACAN, COL2A, SOX9, and MMP13 expression in chondrocytes. ( B ) Relative mRNA expression levels of ACAN, COL2A, SOX9, and MMP13 in chondrocytes. Statistical significance was determined using one-way ANOVA for multiple group comparisons, followed by post hoc tests. All data are presented as mean ± SD (n = 3). **P < 0.01, ***P < 0.001, and ns = no statistically significant difference between groups. ( C ) Alcian blue staining images of chondrocytes. ( D ) Immunofluorescence images of COL2A in chondrocytes. ( E ) Immunofluorescence images of MMP13 in chondrocytes.

Article Snippet: Overnight incubation at 4°C with diluted primary antibodies followed: ACAN (1:2000, Cat# 68350-1-Ig, Proteintech, USA), COL2A (1:2000, Cat# 28459-1-AP, Proteintech, USA), SOX9 (1:2000, Cat# 67439-1-Ig, Proteintech, USA), MMP13 (1:2000, Cat# 18165-1-AP, Proteintech, USA), CD9 (1:1000, Cat# 13174, CST, USA), CD63 (1:1000, Cat# 52090, CST, USA), GM130 (1:1000, Cat# 12480, CST, USA), Bcl-2 (1:1000, Cat# 3498, CST, USA), P53 (1:1000, Cat# 9282, CST, USA), and GAPDH (1:10000, Cat# 10494-1-AP, Proteintech, USA).

Techniques: Expressing, Western Blot, Staining, Immunofluorescence

Journal: Aging and Disease

Article Title: Enhanced Chondrogenic Potential and Osteoarthritis Treatment Using Cyaonoside A-Induced MSC Delivered via a Hyaluronic Acid-Based Hydrogel System

doi: 10.14336/AD.2024.10016

Figure Lengend Snippet: In vivo therapeutic evaluation of hydrogel loaded with CyA-induced MSC . ( A ) Representative 3D reconstruction images of joints in different groups. ( B ) H&E staining of articular cartilage in different groups. ( C ) S&F staining of articular cartilage in different groups. ( D ) Immunohistochemical staining of COL2A in articular cartilage across different groups. ( E ) Immunofluorescence staining of Caspase-3 in articular cartilage across different groups. (F, G) Relative mRNA expression levels of ACAN and COL2A in joint tissues of different groups. Statistical significance was determined using one-way ANOVA for multiple group comparisons, followed by post hoc tests. All data are presented as mean ± SD (n = 3). *p < 0.05, **p < 0.01, ***p < 0.001, and ns = no statistically significant difference between groups.

Article Snippet: Overnight incubation at 4°C with diluted primary antibodies followed: ACAN (1:2000, Cat# 68350-1-Ig, Proteintech, USA), COL2A (1:2000, Cat# 28459-1-AP, Proteintech, USA), SOX9 (1:2000, Cat# 67439-1-Ig, Proteintech, USA), MMP13 (1:2000, Cat# 18165-1-AP, Proteintech, USA), CD9 (1:1000, Cat# 13174, CST, USA), CD63 (1:1000, Cat# 52090, CST, USA), GM130 (1:1000, Cat# 12480, CST, USA), Bcl-2 (1:1000, Cat# 3498, CST, USA), P53 (1:1000, Cat# 9282, CST, USA), and GAPDH (1:10000, Cat# 10494-1-AP, Proteintech, USA).

Techniques: In Vivo, Staining, Immunohistochemical staining, Immunofluorescence, Expressing