Three-dimensional adipose-derived stem cell spheroids exert potent therapeutic effects on rat femur osteochondral defects

•Adipose-derived stem cell has therapeutic effect on osteochondral defect of rat’s knee.•3D culture of adipose-derived stem cell is more effective in treatment than 2D culture.•Spheroids are an effective form of three-dimensional culture of adipose-derived stem cell. Spheroids can allow three-dimensional (3D) cell culture without scaffolds, potentially promoting the production of growth factors from adipose-derived stem cells (ADSCs). We hypothesized that ADSC spheroids exert more favourable effects on osteochondral defects than ADSCs in two-dimensional (2D) cultures. The purpose of this study was to compare the therapeutic effects of 2D and 3D cultures of ADSCs on osteochondral defects using animal models. Rat femoral osteochondral defects were created. When creating osteochondral defects, phosphate‐buffered saline, 2D ADSCs, or ADSC spheroids as a 3D culture were administered on to the lesion. At 2, 4, 6, 8, 10 and 12 weeks post-surgery, knee tissues were harvested and evaluated via histological examination. The expression of genes related to growth factors and apoptosis were compared between 2D and 3D ADSCs. Histologically, the repair of osteochondral defects was significantly enhanced in 3D ADSCs than in 2D ADSCs in terms of the Wakitani score and cartilage repair rate. In 3D ADSCs, TGF-β1, VEGF, HGF and BMP-2 were significantly upregulated, while apoptosis was suppressed in the early phase. The therapeutic effects of 3D ADSC spheroids on osteochondral defects were more potent than those of 2D ADSCs. The upregulated expression of growth factors and suppression of apoptosis could contribute to promoting these therapeutic effects. Overall, ADSC spheroids can help treat osteochondral defects.