Journal: Scientific Reports

Article Title: BMP7 reduces the fibrocartilage chondrocyte phenotype

doi: 10.1038/s41598-021-99096-0

Figure Lengend Snippet: BMP7 increases MMP2 expression and activity in SW1353 cells. ( A ) SW1353 cells (biological triplicates) were exposed to 1 nM BMP7 for 24 h and MMP2 mRNA expression was determined using RT-qPCR analysis. Data were normalized to cyclophillin expression and set relative to control conditions. ( B / C ) In parallel acquired samples with identical treatment (n = 3) MMP2 protein levels and activity in culture supernatant were measured using a MMP2 ELISA and MMP2 activity assay (relative to control condition). ( D ) OA-HACs (n = 18) were exposed to 1 nM BMP7 for 24 h after which MMP2 mRNA expression was measured by RT-qPCR. Data were normalized for cyclophilin mRNA levels and set relative to control conditions per patient. ( E ) SW1353 cells (biological quadruplicates) were pre-incubated with the selective MMP2 inhibitor OA-Hy (50 µM) for 24 h. MMP2 activity was determined and calculated relative to the control condition. ( F ) Collagen type I protein levels were detected in SW1353 cells (biological quintiplicates) by immunocytochemistry in control conditions and conditions exposed to MMP2 inhibitor OA-Hy (50 µM) with and without BMP7. Data were normalized for DNA content and set relative to control conditions. Statistical significance was determined using 2-tailed unpaired Student’s t-tests ( D per donor and as a group). Bars show the mean (± SEM). * P < 0.05, ** P < 0.01, *** P < 0.001 versus control conditions.

Article Snippet: SW1353 chondrosarcoma cells and OA HACs were treated with 1 nM BMP7 (R&D Systems, Minneapolis, MN, USA) for 24 h. Prior to BMP7 exposure, SW1353 cells were pre-incubated for 1 h with 50 μM selective MMP2 Inhibitor I (OA-Hy SantaCruz Biotechnology, Dallas, TX, USA).

Techniques: Expressing, Activity Assay, Quantitative RT-PCR, Control, Enzyme-linked Immunosorbent Assay, Incubation, Immunocytochemistry

Journal: Scientific Reports

Article Title: BMP7 reduces the fibrocartilage chondrocyte phenotype

doi: 10.1038/s41598-021-99096-0

Figure Lengend Snippet: Proposed molecular interactions by which BMP7 attenuates the fibrocartilage chondrocyte phenotype. The potential molecular pathway by which BMP7 attenuates the fibrocartilage chondrocyte phenotype as proposed in this study. Upon BMP7 exposure, chondrocytic cells demonstrate reduced SMAD3 signaling, interfering with PAI1 expression and activity. In addition, following BMP7 exposure, MMP2 expression and activity is increased via PAI1 and MT1-MMP, resulting in a reduced fibrocartilage chondrocyte phenotype. BMP7’s mode of action to reduce chondrocyte fibrosis is MMP2-dependent. Dashed lines: interactions which have been found in organ-fibrosis (with corresponding references), but have not been demonstrated directly in the present study. Solid lines: interactions directly demonstrated in the present study (regarding chondrocyte fibrosis).

Article Snippet: SW1353 chondrosarcoma cells and OA HACs were treated with 1 nM BMP7 (R&D Systems, Minneapolis, MN, USA) for 24 h. Prior to BMP7 exposure, SW1353 cells were pre-incubated for 1 h with 50 μM selective MMP2 Inhibitor I (OA-Hy SantaCruz Biotechnology, Dallas, TX, USA).

Techniques: Expressing, Activity Assay

Journal: Cancer medicine

Article Title: Anti-tumor effects of a nonsteroidal anti-inflammatory drug zaltoprofen on chondrosarcoma via activating peroxisome proliferator-activated receptor gamma and suppressing matrix metalloproteinase-2 expression.

doi: 10.1002/cam4.1438

Figure Lengend Snippet: Figure 3. (A) Cell migration assay was carried out using PPARγ knockdown cell lines, OUMS27shPPAR and SW1353shPPAR, and the control cell lines, OUMS27shNTC and SW1353shNTC with or without the treatment of zaltoprofen. A selective inhibitor of MMP2, ARP100, was used at 25 μmol/L. Scale bar = 100 μm. (B, C) The fielded wound area (%) was calculated in OUMS27 (B) and SW1353 cells (C). Values represented as the mean ± SEM. Asterisks denote a statistically significant difference (*P < 0.05 and **P < 0.01).

Article Snippet: Zaltoprofen and ARP100, a selective MMP2 inhibitor, were purchased from Santa Cruz (Santa Cruz Biotechnology, Inc., Santa Cruz, CA) and dissolved in dimethyl sulfoxide (DMSO).

Techniques: Cell Migration Assay, Knockdown, Control

Journal: Cancer medicine

Article Title: Anti-tumor effects of a nonsteroidal anti-inflammatory drug zaltoprofen on chondrosarcoma via activating peroxisome proliferator-activated receptor gamma and suppressing matrix metalloproteinase-2 expression.

doi: 10.1002/cam4.1438

Figure Lengend Snippet: Figure 4. (A) Cell invasion assay was carried out using a PPARγ knockdown cell line SW1353shPPAR and the control cell line SW1353shNTC with or without the treatment of zaltoprofen. A selective inhibitor of MMP2, ARP100, was used at 25 μmol/L. Scale bar = 100 μm. (B) Relative invaded cell number was calculated. Values represented as the mean ± SEM. Asterisks denote a statistically significant difference (**P < 0.01). (C) Gelatin zymography. Effects of zaltoprofen (400 μmol/L) on MMP2 activities in PPARγ knockdown cell lines, OUMS27shPPAR and SW1353shPPAR, and the control cell lines, OUMS27shNTC and SW1353shNTC.

Article Snippet: Zaltoprofen and ARP100, a selective MMP2 inhibitor, were purchased from Santa Cruz (Santa Cruz Biotechnology, Inc., Santa Cruz, CA) and dissolved in dimethyl sulfoxide (DMSO).

Techniques: Invasion Assay, Knockdown, Control, Zymography

Journal: Cancer medicine

Article Title: Anti-tumor effects of a nonsteroidal anti-inflammatory drug zaltoprofen on chondrosarcoma via activating peroxisome proliferator-activated receptor gamma and suppressing matrix metalloproteinase-2 expression.

doi: 10.1002/cam4.1438

Figure Lengend Snippet: Figure 6. Histopathological findings of the tumors at the second operation (preadministration of zaltoprofen, Pre) and the final operation (postadministration of zaltoprofen, Post). Scale bar = 25 μm. (A) HE staining. (B, C) Immunofluorescent stainings for PPARγ (B) and MMP2 (C). DAPI, a nuclear staining.

Article Snippet: Zaltoprofen and ARP100, a selective MMP2 inhibitor, were purchased from Santa Cruz (Santa Cruz Biotechnology, Inc., Santa Cruz, CA) and dissolved in dimethyl sulfoxide (DMSO).

Techniques: Staining