Journal: Journal of Translational Autoimmunity

Article Title: T cell proliferative response to a homocitrullinated peptide correlates with joint pathology in collagen induced arthritis

doi: 10.1016/j.jtauto.2025.100345

Figure Lengend Snippet: RA-Specific Serum IgG Antibodies in DBA/1J Mice. Sera were collected biweekly from DBA/1J mice immunized with bovine type II collagen (bCII) (arthritic: N = 9–18; non-arthritic: N = 17–37), or PBS (N = 14) and tested via indirect ELISA for A) anti-CitP, and B) anti-HomoCitP IgG antibodies. Graphs show the median [IQR] for each group, with a cut-off indicating the limit of detection (dashed line). Statistical analysis was performed using a mixed effects model with the Geisser-Greenhouse correction and Tukey's multiple comparisons test. The resulting p-values are indicated on the graphs.

Article Snippet: The anti-mCII ELISA kits were used according to manufacturer's protocol (2036T; Chondrex).

Techniques: Indirect ELISA

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: 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: The COL2A primary antibody (1:1000, Cat# 28459-1-AP, Proteintech, USA) was applied, and the sections were incubated overnight at 4°C.

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: The COL2A primary antibody (1:1000, Cat# 28459-1-AP, Proteintech, USA) was applied, and the sections were incubated overnight at 4°C.

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: The COL2A primary antibody (1:1000, Cat# 28459-1-AP, Proteintech, USA) was applied, and the sections were incubated overnight at 4°C.

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

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