Due to the poor self-regenerative capacity of native cartilage, the treatment of articular cartilage defects is a significant clinical challenge. The cartilage tissue engineered scaffolds with high chondrogenic capacity might provide an efficient strategy for the repair of cartilage defects. Herein, we report an aligned porous poly (L-lactic acid) (PLLA) electrospun fibrous membrane with a biomimetic surface for the rapid regeneration of cartilage tissue. The PLLA electrospun fibers aligned in a single direction and generated ellipse-shaped nanopores in situ onto the surface of the fibers. Subsequently, chondroitin sulfate (CS) was coated on the surface of the fibers using polydopamine (PDA) as an adhesive polymeric bridge. The in vitro results revealed that the aligned porous composite membranes with a biomimetic surface (PL/PDA/CS) could stimulate the attachment and proliferation of chondrocytes and rabbit bone marrow mesenchymal stem cells (rBMSCs), as well as inducing chondrogenic gene expression in rBMSCs. Furthermore, the PL/PDA/CS membranes significantly facilitated the filling of defects and the regeneration of hyaline cartilage-like tissue in vivo. The results suggested that the hierarchical aligned porous fibrous electrospun membrane with a biomimetic surface modification is a potential therapeutic strategy for highly efficient cartilage defect repair.