AbstractOsteochondral defects affect both the articular cartilage and the underlying subchondral bone, but poor osteochondral regeneration is still a daunting challenge. Although the tissue engineering technology provides a promising approach for osteochondral repair, an ideal biphasic scaffold is in high demand with regards to proper biomechanical strength. In this study, an oriented poly(l‐lacticacid)‐co‐poly(ε‐caprolactone) P(LLA‐CL)/collagen type I(Col‐I) nanofiber yarn mesh, fabricated by dynamic liquid electrospinning served as a skeleton for a freeze‐dried Col‐I/Hhyaluronate (HA) chondral phase (SPONGE) to enhance the mechanical strength of the scaffold. In vitro results show that the Yarn Col‐I/HA hybrid scaffold (Yarn‐CH) can allow the cell infiltration like sponge scaffolds. Using porous beta‐tricalcium phosphate (TCP) as the osseous phase, the Yarn‐CH/TCP biphasic scaffold was then assembled by freeze drying. After combination of bone marrow mesenchymal stem cells, the biphasic complex was successfully used to repair the osteochondral defects in a rabbit model with greatly improved repairing scores and compressive modulus. © 2014 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 103A: 581–592, 2015.