odanacatib  (MedChemExpress)

Journal: Frontiers in Endocrinology

Article Title: CU Cilia – an application for image analysis by machine learning – reveals significance of cysteine cathepsin K activity for primary cilia of human thyroid epithelial cells

doi: 10.3389/fendo.2025.1588394

Figure Lengend Snippet: Detection of nuclei and primary cilia of human thyroid epithelial cells in vitro using CellProfiler™ pipelines. (A) Bar charts comparing the numbers of detected nuclei and primary cilia (a1) , cilia frequencies (a2) and cilia lengths (a3) as revealed by CellProfiler™ pipelines set up by non-expert (pink, left panels) and expert users (violet, right panels). Mean values ± standard deviations are displayed in the bar charts with individual data points indicated by circles or dots (a1–a3) . Statistical analysis was by Kruskal-Wallis multiple comparisons tests; levels of significance are indicated as **** for p<0.0001. Note that the ranges of measurements differed between non-expert and expert determinations, while the 1 h-treatments with broad-spectrum (E64d) or specific cysteine peptidase inhibitors (CA074me, Odanacatib, CathLi III) did not usually differ from controls (DMSO). (B) Merged channel confocal laser scanning micrographs depicting ARL13B-positive primary cilia ( B , green in b1–b5 ; black in b1’–b5’ ) and Draq5™-stained nuclei ( B , blue in b1–b5 ) of DMSO-treated controls (b1, b1’) or Nthy-ori 3–1 cell cultures treated with broad-spectrum (b2, b2’) or specific inhibitors of cathepsin B (b3, b3’) , cathepsin K (b4, b4’) or cathepsin L (b5, b5’) , respectively. Note that corresponding single channels of anti-ARL13B-positive primary cilia are shown in inverted contrast (b1’–b5’) for clarity. Scale bars represent 50 µm. Identical images were analyzed with the two different pipelines with n=9 technical replicates, except n=8 for Odanacatib-treated cells.

Article Snippet: However, in this study, cell cultures were incubated for 1 h and 24 h with 10 μM E64d (#BML-PI107, Enzo Life Sciences, Lörrach, Germany) for broad-spectrum inhibition of cysteine peptidase activities and with cysteine cathepsin B-, K-, and L-specific inhibitors, namely, 10 μM CA074 me (#BML-PI126, Enzo Life Sciences, Lörrach, Germany), 10 μM Odanacatib (#S1115, Selleck Chemicals GmbH, Cologne, Germany), and 10 μM cathepsin L inhibitor III (#219427, Merck KGaA, Darmstadt, Germany), respectively.

Techniques: In Vitro, Staining

Journal: Frontiers in Endocrinology

Article Title: CU Cilia – an application for image analysis by machine learning – reveals significance of cysteine cathepsin K activity for primary cilia of human thyroid epithelial cells

doi: 10.3389/fendo.2025.1588394

Figure Lengend Snippet: Elongation of primary cilia extending from human thyroid epithelial cells upon treatment with cysteine peptidase inhibitors for 24 h as revealed by CellProfiler™ and CU Cilia analyses. (A) Bar charts comparing the numbers of detected nuclei and primary cilia (a1) , cilia frequencies (a2) and cilia lengths (a3) as revealed by CellProfiler™ pipelines set up by an expert user (violet, left panels) and CU Cilia (green, right panels). Identical images (n=9) were analyzed with both approaches. Mean values ± standard deviations are displayed in the bar charts with individual data indicated by circles or dots (a1–a3) . Note that both approaches yielded comparable results. Cilia elongation was observed upon treatment with E46d and Odanacatib for 24 h (a3) . Statistical analysis was by Kruskal-Wallis and Dunn’s multiple comparisons tests; levels of significance are indicated as * for p<0.01, ** for p<0.001, and **** for p<0.0001. (B) Merged channel confocal laser scanning micrographs depicting ARL13B-positive primary cilia ( B , green in b1–b5 ; black in b1’–b5’ ) and Draq5™-stained nuclei ( B , blue in b1–b5 ) of DMSO-treated controls (b1, b1’) or Nthy-ori 3–1 cell cultures treated with broad-spectrum (b2, b2’) or specific inhibitors of cathepsin B (b3, b3’) , cathepsin K (b4, b4’) or cathepsin L (b5, b5’) , respectively. Note that corresponding single channels of anti-ARL13B-positive primary cilia are shown in inverted contrast (b1’–b5’) for clarity. Scale bars represent 50 µm. Cathepsin L inhibitor III treatment for 24 h was cytotoxic towards Nthy-ori 3–1 cells as obvious from fewer detected nuclei (a1) and more abundant apoptotic bodies (b5) .

Article Snippet: However, in this study, cell cultures were incubated for 1 h and 24 h with 10 μM E64d (#BML-PI107, Enzo Life Sciences, Lörrach, Germany) for broad-spectrum inhibition of cysteine peptidase activities and with cysteine cathepsin B-, K-, and L-specific inhibitors, namely, 10 μM CA074 me (#BML-PI126, Enzo Life Sciences, Lörrach, Germany), 10 μM Odanacatib (#S1115, Selleck Chemicals GmbH, Cologne, Germany), and 10 μM cathepsin L inhibitor III (#219427, Merck KGaA, Darmstadt, Germany), respectively.

Techniques: Staining

Journal: Frontiers in Endocrinology

Article Title: CU Cilia – an application for image analysis by machine learning – reveals significance of cysteine cathepsin K activity for primary cilia of human thyroid epithelial cells

doi: 10.3389/fendo.2025.1588394

Figure Lengend Snippet: Elongation and thinning of primary cilia extending from human thyroid epithelial cells upon treatment with general cysteine peptidase and cathepsin K-specific inhibitors for 24 h as revealed by advanced CU Cilia analyses. (A) Bar charts and box plots comparing the areas of primary cilia (a1) , major axis length (a2) , minor axis length (a3) , cilia perimeters (a4) , cilia skeleton lengths (a5) , mean distances of cilia to nearest center of mass (a6) or to nuclei boundaries (a7) as well as form factor (a8) or eccentricity of primary cilia (a9) as revealed by CU Cilia. Images (n=6) of DMSO-treated controls or Nthy-ori 3–1 cell cultures treated for 24 h with broad-spectrum or specific inhibitors of cathepsin B, cathepsin K, or cathepsin L, respectively, were analyzed and data is denoted as indicated. Mean values ± standard deviations are displayed in the bar charts with individual data indicated by circles or dots (a1–a5, a8–a9) , while box plots are displayed in a6 and a7 . Statistical analysis was conducted by Kruskal-Wallis and Dunn’s multiple comparisons tests; levels of significance are indicated as ** for p<0.01, *** for p<0.001, and **** for p<0.0001. Cilia elongation (a2, a4, a5) and cilia thinning (a3) was observed upon treatment with CA074me, E46d and Odanacatib for 24 h, while more elliptic cilia were observed in E64d- and Odanacatib-treated Nthy-ori 3–1 cells, only (a8, a9) . (B) Output images generated by CU Cilia consisting of merged channel confocal laser scanning micrographs depicting ARL13B-positive primary cilia ( B , white) and Draq5™-stained nuclei ( B , blue) with identified objects boxed ( B , orange) and object sizes indicated in white font (B) of Nthy-ori 3–1 control cell cultures. Schematic drawing summarizing the advanced features of CU Cilia measurements.

Article Snippet: However, in this study, cell cultures were incubated for 1 h and 24 h with 10 μM E64d (#BML-PI107, Enzo Life Sciences, Lörrach, Germany) for broad-spectrum inhibition of cysteine peptidase activities and with cysteine cathepsin B-, K-, and L-specific inhibitors, namely, 10 μM CA074 me (#BML-PI126, Enzo Life Sciences, Lörrach, Germany), 10 μM Odanacatib (#S1115, Selleck Chemicals GmbH, Cologne, Germany), and 10 μM cathepsin L inhibitor III (#219427, Merck KGaA, Darmstadt, Germany), respectively.

Techniques: Generated, Staining, Control

Journal: Clinical Oral Investigations

Article Title: Cytokine expression of soft tissue cells cultured with titanium discs and their respective supernatants in vitro

doi: 10.1007/s00784-024-06123-1

Figure Lengend Snippet: Workflow represents the sample distribution of 216 titanium discs in three groups of surface treatment: SLA, SBF and SBF + ODN. The SLA was considered as a control group. The diagram describes the analyses performed and the respective exposure times for each experiment

Article Snippet: The discs were immersed and incubated in SBF containing either no or 260 µg/mL of odanacatib (ODN, MedChemExpress LLC, Monmouth Junction, NJ) at pH 7.25 and 37 °C for 4 days.

Techniques: Control

Journal: Clinical Oral Investigations

Article Title: Cytokine expression of soft tissue cells cultured with titanium discs and their respective supernatants in vitro

doi: 10.1007/s00784-024-06123-1

Figure Lengend Snippet: Scanning electron microscopy (SEM) images of the surfaces of SLA, SBF, and SBF + ODN discs at 10,000 × magnification. A - D SLA shows uniform microroughness with varying depths and sizes. E – H SBF exhibits the expected spider web-like pores of the hydroxyapatite coating. I - L SBF + ODN displays a rough texture with globular microcavities, interlocking pores, and nanometric particles. Obviously, the presence of ODN affects the morphology of the hydroxyapatite coating compared to the SBF alone discs – and its wettability

Article Snippet: The discs were immersed and incubated in SBF containing either no or 260 µg/mL of odanacatib (ODN, MedChemExpress LLC, Monmouth Junction, NJ) at pH 7.25 and 37 °C for 4 days.

Techniques: Electron Microscopy

Journal: Clinical Oral Investigations

Article Title: Cytokine expression of soft tissue cells cultured with titanium discs and their respective supernatants in vitro

doi: 10.1007/s00784-024-06123-1

Figure Lengend Snippet: Energy-dispersive X-ray (EDX) spectroscopy of the surfaces of SLA, SBF, and SBF + ODN discs, showing the apparent concentration of chemical elements present on the surface

Article Snippet: The discs were immersed and incubated in SBF containing either no or 260 µg/mL of odanacatib (ODN, MedChemExpress LLC, Monmouth Junction, NJ) at pH 7.25 and 37 °C for 4 days.

Techniques: Spectroscopy, Concentration Assay

Journal: Clinical Oral Investigations

Article Title: Cytokine expression of soft tissue cells cultured with titanium discs and their respective supernatants in vitro

doi: 10.1007/s00784-024-06123-1

Figure Lengend Snippet: Variation in contact angle values (θ) in different surface modifications

Article Snippet: The discs were immersed and incubated in SBF containing either no or 260 µg/mL of odanacatib (ODN, MedChemExpress LLC, Monmouth Junction, NJ) at pH 7.25 and 37 °C for 4 days.

Techniques:

Journal: Clinical Oral Investigations

Article Title: Cytokine expression of soft tissue cells cultured with titanium discs and their respective supernatants in vitro

doi: 10.1007/s00784-024-06123-1

Figure Lengend Snippet: Surface roughness was evaluated by measuring Ra ( A ) and Rz ( B ) values for each group SLA, SBF and SBF + ODN. The results showed that the ODN coating significantly increased surface roughness compared to SBF alone. Different symbol shapes represent mean values of independent readings. Statistical analysis was performed using ratio-paired t-tests compared to untreated controls, and p-values are shown

Article Snippet: The discs were immersed and incubated in SBF containing either no or 260 µg/mL of odanacatib (ODN, MedChemExpress LLC, Monmouth Junction, NJ) at pH 7.25 and 37 °C for 4 days.

Techniques:

Journal: Clinical Oral Investigations

Article Title: Cytokine expression of soft tissue cells cultured with titanium discs and their respective supernatants in vitro

doi: 10.1007/s00784-024-06123-1

Figure Lengend Snippet: Cell viability assays were performed on human gingival fibroblasts and HSC2 cells stimulated with the supernatants and discs from each group—SLA, SBF, and SBF + ODN overnight

Article Snippet: The discs were immersed and incubated in SBF containing either no or 260 µg/mL of odanacatib (ODN, MedChemExpress LLC, Monmouth Junction, NJ) at pH 7.25 and 37 °C for 4 days.

Techniques:

Journal: Clinical Oral Investigations

Article Title: Cytokine expression of soft tissue cells cultured with titanium discs and their respective supernatants in vitro

doi: 10.1007/s00784-024-06123-1

Figure Lengend Snippet: Cathepsin K activity was conducted with titanium discs coated with SBF and SBF + ODN, in the presence of osteoclast lysates. Fluorescence intensity indicating CATK activity was determined. Statistical analysis was performed using ratio-paired t-tests. Notably, the presence of ODN significantly lowers the CATK activity intrinsic to the osteoclast lysate

Article Snippet: The discs were immersed and incubated in SBF containing either no or 260 µg/mL of odanacatib (ODN, MedChemExpress LLC, Monmouth Junction, NJ) at pH 7.25 and 37 °C for 4 days.

Techniques: Activity Assay, Fluorescence

Journal: Clinical Oral Investigations

Article Title: Cytokine expression of soft tissue cells cultured with titanium discs and their respective supernatants in vitro

doi: 10.1007/s00784-024-06123-1

Figure Lengend Snippet: Bone marrow-derived cells were exposed to supernatants ( A , C ) and titanium discs ( B , C ) from each group in the presence of M-CSF, RANKL, and TGF-β (MRT). A , B Gene expression levels of TRAP and CTSK were normalized to untreated cells. Different symbol shapes in the graphs represents independent experiments. The SBF coating resulted in decreased TRAP and CTSK expression, but this was not reflected in TRAP staining. Statistical analysis was performed using ratio-paired t-tests, with p-values indicated. C Representative TRAP-stained images show that osteoclast formation was not significantly suppressed by SLA, SBF, or SBF + ODN disc and respective supernatants

Article Snippet: The discs were immersed and incubated in SBF containing either no or 260 µg/mL of odanacatib (ODN, MedChemExpress LLC, Monmouth Junction, NJ) at pH 7.25 and 37 °C for 4 days.

Techniques: Derivative Assay, Expressing, Staining