Method Profile
Miniaturized human cartilage pellets support dose/time screening of thyroid-receptor antagonists, followed by a native osteochondral tissue comparison.
Assess candidate effects on T3-induced terminal maturation in osteoarthritis-related human cartilage models.
Primary articular chondrocytes from preserved cartilage of joint-replacement patients form 96-well neo-cartilage organoids; osteochondral explants retain cartilage with underlying bone.
T3-induced maturation followed by candidate compound dose/time comparisons; gene expression and secreted MMP3 are measured separately in each model.
Human material
Macroscopically preserved cartilage from osteoarthritis surgical specimens; native explants and newly formed cartilage organoids are distinct tissue states.
Perturbations
T3 at 100 ng/mL; Galapagos-developed thyroid hormone receptor antagonists A, B and C. Organoid screening uses 0.3, 1, 3 and 10 μM for 4 or 7 days.
Endpoints
RT-qPCR for terminal/early maturation and cartilage-state markers; MMP3 ELISA in culture medium.
Evaluate markers within each tissue model, retain dose and duration distinctions and use the explant experiment to examine whether a screening response extends to native tissue.
Establish T3 responses in both systems, screen three compounds in organoids, and compare A/B responses in explants.
Dose/time screening: one donor with 1–2 samples per condition. Explant compound figure: 2–5 donors and 1–2 samples; these are different experiments from broader model-characterization sets.
Untreated and T3-induced conditions, candidate doses/times and the two tissue formats.
Organoid Results report dose-dependent CCDC80 reduction, strongest at 10 μM/4 days. Compound A restored several terminal-maturation markers and MMP3 toward baseline in explants; B did not restore MMP3, while ATOH8/COL10A1 were unchanged.
A 96-well cartilage organoid screen is followed by human osteochondral explant testing. Compound A most consistently reverses selected T3-related changes, while small screening samples and different marker responses constrain interpretation.
