A　double-layered　collagen/silk　fibroin　composite　scaffold　that　incorporates　TGF-β1/poly-L-lysine　nanoparticles　was　prepared　in　this　study.　The　microstructure　and　morphology　of　the　obtained　scaffold　were　examined　through　transmission　electron　microscopy　and　scanning　electron　microscopy.　Scaffold　exhibited　a　double-layered　structure　with　TGF-β1-loaded　poly-L-lysine-heparin　(PLL-Hep)　nanoparticles　sandwiched　between　dense　and　loose　layers.　TGF-β1　release　displayed　a　spatiotemporal　control　model.　In　vitro　biocompatibility　and　fluorescence　staining　test　with　Bone　Marrow　Stromal　Cells　revealed　that　scaffold　could　significantly　enhance　cell　proliferation.　The　prepared　scaffold　was　then　implanted　into　full-thickness　articular　cartilage　defects　in　rabbits.　The　results　indicated　that　the　group　exhibiting　loose　layer　facing　the　surface　of　subchondral　bone　was　repaired　more　effectively　than　displayed　a　loose　layer　facing　the　opposite　direction.　Therefore,　this　composite　is　expected　to　be　a　potential　scaffold　for　cartilage　tissue　engineering　material.　©　2015　American　Scientific　Publishers.　All　rights　reserved.