matrix metalloproteinase mmp 13  (Proteintech)

Journal: Materials Today Bio

Article Title: Cuprorivaite microspheres inhibit cuproptosis and oxidative stress in osteoarthritis via Wnt/β-catenin pathway

doi: 10.1016/j.mtbio.2024.101300

Figure Lengend Snippet: Anti-inflammatory role of Cuprorivaite (CaCuSi 4 O 10 ) microspheres in IL-1β-stimulated chondrocytes. (A) Cell counting kit-8 test for cell proliferation. (B) Fluorescein diacetate for cell viability detection. (C) Cell apoptosis by flow cytometry. (D) ELISA detection for TNF-α, IL-6 and MMP13 in cell supernatant. (E) Western blot detection for TNF-α, IL-6 and MMP13 in cell lysates. (F) RT-qPCR quantification for in TNF-α, IL-6 and MMP13. ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001.

Article Snippet: Then, levels of TNF-α, IL-6, and MMP13 in cell supernatant and serum samples were detected by the multi-functional microplate detector, with mouse TNF-α (CSB-E04741m, CUSABIO, Wuhan, China), mouse IL-6 (CSB-E04639m, CUSABIO, Wuhan, China) and mouse MMP13 (CSB-E07413m, CUSABIO, Wuhan, China) kit.

Techniques: Cell Counting, Flow Cytometry, Enzyme-linked Immunosorbent Assay, Western Blot, Quantitative RT-PCR

Journal: Materials Today Bio

Article Title: Cuprorivaite microspheres inhibit cuproptosis and oxidative stress in osteoarthritis via Wnt/β-catenin pathway

doi: 10.1016/j.mtbio.2024.101300

Figure Lengend Snippet: Cuprorivaite (CaCuSi 4 O 10 ) microspheres improved IL-1β-induced injury in chondrocytes via inhibiting Wnt/β-catenin pathway. (A) Cell counting kit-8 test for cell viability. (B) Fluorescein diacetate for cell viability detection. (C) Cell apoptosis by flow cytometry. (D) ELISA detection for TNF-α, IL-6 and MMP13 in cell supernatant. (E) Western blot detection for TNF-α, IL-6 and MMP13 in cell lysates. (F) RT-qPCR quantification for extracellular matrix components including collagen II and SOX9. (G) Intracellular copper content. (H) RT-qPCR quantification for cuproptosis biomarkers including ATP7B and FDX1. (I) Western blot detection for ATP7B and FDX1. (J) Oxidative stress detection including MDA, SOD and GSH. (K) RT-qPCR quantification for Wnt1, GSK3β and β-catenin. ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001.

Article Snippet: Then, levels of TNF-α, IL-6, and MMP13 in cell supernatant and serum samples were detected by the multi-functional microplate detector, with mouse TNF-α (CSB-E04741m, CUSABIO, Wuhan, China), mouse IL-6 (CSB-E04639m, CUSABIO, Wuhan, China) and mouse MMP13 (CSB-E07413m, CUSABIO, Wuhan, China) kit.

Techniques: Cell Counting, Flow Cytometry, Enzyme-linked Immunosorbent Assay, Western Blot, Quantitative RT-PCR

Journal: Materials Today Bio

Article Title: Cuprorivaite microspheres inhibit cuproptosis and oxidative stress in osteoarthritis via Wnt/β-catenin pathway

doi: 10.1016/j.mtbio.2024.101300

Figure Lengend Snippet: Validation of mechanism of Cuprorivaite (CaCuSi 4 O 10 ) microspheres in OA. (A) Hematoxylin and eosin staining. (B) Safranin-O staining/fast green staining. (C) OARSI score. (D) ELISA detection for TNF-α, IL-6 and MMP13 in the serum. (E) Western blot detection for TNF-α, IL-6 and MMP13 in cartilage tissue. (F) RT-qPCR quantification for extracellular matrix components including collagen II and SOX9 in cartilage tissue. (G) Copper content in cartilage tissue. (H) RT-qPCR quantification for cuproptosis biomarkers including ATP7B and FDX1 in cartilage tissue. (I) Western blot detection for ATP7B and FDX1 in cartilage tissue. (J) Representative images of FDX1 expression in cartilage tissue detected by immunohistochemistry. (K) Oxidative stress detection including MDA, SOD and GSH in cartilage tissue. (L) RT-qPCR quantification for Wnt1, GSK3β and β-catenin in cartilage tissue. ∗∗∗ P < 0.001.

Article Snippet: Then, levels of TNF-α, IL-6, and MMP13 in cell supernatant and serum samples were detected by the multi-functional microplate detector, with mouse TNF-α (CSB-E04741m, CUSABIO, Wuhan, China), mouse IL-6 (CSB-E04639m, CUSABIO, Wuhan, China) and mouse MMP13 (CSB-E07413m, CUSABIO, Wuhan, China) kit.

Techniques: Biomarker Discovery, Staining, Enzyme-linked Immunosorbent Assay, Western Blot, Quantitative RT-PCR, Expressing, Immunohistochemistry

Journal: Scientific Reports

Article Title: Interferon-gamma signaling promotes cartilage regeneration after injury

doi: 10.1038/s41598-024-58779-0

Figure Lengend Snippet: Genetic ablation of IFN-γR1 exacerbated OA pathogenesis in mice after DMM. ( A ) Representative images of safranin-O/fast green-stained sections of the knee joints of WT and IFN-γR1 −/− mice after DMM. Boxed areas in the right panel are shown at higher magnification. Scale bar = 50 μm. ( B ) Representative images of the synovium in the knee joint of each group. Boxed areas in the right panel are shown at higher magnification. Arrows indicate increased synovial cell proliferation. Scale bar = 50 μm. ( C ) Quantitative analyses of the Osteoarthritis Research Society International (OARSI), synovitis, subchondral bone, and osteophyte formation scores after DMM in WT and IFN-γR1 −/− mice (n = 5–7). ( D , E ) Representative images of immunohistochemical staining for aggrecan ( D ) and matrix metalloproteinase (MMP)-13 ( E ) expression in knee sections of WT and IFN-γR1 −/− mice after DMM. Quantification of aggrecan- and MMP-13-positive chondrocytes in the articular cartilage of sham-operated and DMM joints (n = 5–6 per group; five sections/mouse). Scale bar = 50 μm. ( F ) Mechanical allodynia was evaluated using the von Frey filament test (left; n = 8–10) and the withdrawal threshold was measured using a pressure algometer (right; n = 6–7) after DMM. Data are shown as mean ± SEM. *P < 0.05, **P < 0.01, ***P < 0.001 versus WT sham or WT DMM according to the two-tailed Mann–Whitney U test ( C ), unpaired two-tailed t -test ( D , E ), and Kruskal–Wallis test with Dunn’s multiple comparison test ( F ).

Article Snippet: For immunohistochemistry, the following primary antibodies were used: anti-type II collagen (Abcam, ab34712, dilution 1:200), anti-aggrecan (Millipore, AB1031, dilution 1:200), and anti-matrix metalloproteinase (MMP)-13 (Abcam, ab39012, dilution 1:200).

Techniques: Staining, Immunohistochemical staining, Expressing, Two Tailed Test, MANN-WHITNEY, Comparison