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Objective The management of chondral defects has long been a challenge since the poor self-healing capacity of articular cartilage.Many efforts using approaches ranging from symptomatic treatment to structural cartilage regeneration have obtained very limited satisfactory results.Cartilage tissue engineering, involves an optimized combination of novel scaffolds, cell sources and growth factors, has emerged as a promising strategy for cartilage regeneration and repair.Methods In the study, based on the 3D scaffold of Poly(3-hydroxybutyrate-co-4-hydroxybutyrate) (P3HB4HB) by electrospinning, the cell behaviors of adipose-derived stem cells (ASCs) and chondrocytes seeded onto the scaffold were evaluated using cell morphologies and the abilities of adhesion, migration and proliferation.Next, TGF-β1/scaffolds with 4∶1 coculture of ASCs and chondrocytes were implanted into the full thickness cartilage defects in rabbit knee for 16 weeks.Results ASCs and chondrocytes seeded onto the scaffolds showed better capacities of adhesion, migration and proliferation in vitro.Importantly, implantation with TGF-β1/scaffolds with delivery of ASCs and chondrocytes revealed desirable in vivo healing outcomes.Conclusion These results demonstrate that ASCs have a great potential in the field of tissue engineering.It is possible that the improvement in ASC-based electrospun 3D P3HB4HB scaffold may ultimately lead to better repair of cartilage injury.