Fig. 1

Workflow of monocyte isolation, and macrophage differentiation and polarization. Approximately 3–5 × 10⁶ PBMCs from early- and late-stage BD patients and healthy controls were seeded into 12-well cell culture plates and allowed to adhere for 2 h in a 5% CO₂ incubator at 37 °C. Adherent cells were differentiated into macrophages in RPMI 1640 media supplemented with 10% FBS and M-CSF, 50 ng/mL for 7 days. For classical and alternative phenotypes, Mψ were incubated for additional 24 h with RPMI + 10% FBS supplemented with IFNγ (20 ng/mL) and LPS (100 ng/mL) or IL-4 (20 ng/mL), respectively