The heterogeneous nature of osteochondral tissue requires the construction of implant with biomimetic gradients. Electrospun fibrous meshes with different designs provide the feasibility in arranging such a kind of gradient structure via layer-by-layer stacking. In this study, a kind of triphasic implant was constructed by layering pre-differentiated cell sheets, which were hold by electrospun poly(L-lactide)/gelatin (PLLA/Gel) fibrous meshes containing hyaluronic acid and chondroitin sulfate for the cartilage layer or hydroxyapatite for the bone layer. As for the calcified interlayer, the bone marrow mesenchymal stromal cells (BMSCs) seeded on PLLA/Gel fibrous mesh was pre-differentiated with a mixed osteoinductive/chondroinductive (1:1) medium. With this gradient construct being implanted in rabbit knee osteochondral defect, it was found that both the cartilage and subchondral bone were regenerated effectively with reproduced tidal line structure. The importance of implants with biomimetic gradients for osteochondral defect repair was confirmed, and cell sheets on electrospun fibrous meshes were flexible for gradient structure construction via the layer-by-layer stacking technology. HighlightsComposite fibrous meshes with tissue-specific components are electrospun.Confluent BMSCs on fibrous meshes are chondrogenically or osteogenically induced.BMSCs hypertrophy is induced with the mixture of chondroinductive and osteoinductive medium.The pre-differentiated cell/mesh complexes are stacked layer-by-layer to form gradient construct.The gradient construct efficiently promotes osteochondral regeneration in rabbit joint.
Keywords: multi-layered; Osteochondral; cell sheet; fibrous mesh; gradient.