A self-assembling peptide nanofiber hydrogel (LKP) was developed to mimic the function of TGF-β1 for cartilage regeneration. LKP hydrogel is simple to synthesize and has good biocompatibility and cartilage-promoting ability. However, it has poor mechanical properties and is prone to fragmentation. To address these issues, a series of injectable hydrogel composite scaffolds (SF-GMA/LKP) were prepared by combining LKP with GMA-modified silk fibroin (SF). The SF-GMA/LKP composite scaffolds can induce in-situ filling of cartilage defects and prolong the action time of LKP through interaction between LKP and SF-GMA. The SF-GMA/LKP10 and SF-GMA/LKP20 scaffolds showed the best effect on neocartilage and subchondral bone reconstruction. The composite hydrogel scaffold can be used for high-quality cartilage repair. The SF-GMA/LKP scaffolds were prepared by mixing SF-GMA with LKP peptide nanofiber hydrogels. The scaffolds were characterized using various techniques, including FTIR, SEM, AFM, and rheological analysis. The scaffolds showed good biocompatibility, with no significant toxicity to BMSCs. The SF-GMA/LKP scaffolds promoted the expression of cartilage-related proteins, such as Col-II, Sox9, and ACAN, indicating their potential for cartilage regeneration. In vivo experiments showed that the SF-GMA/LKP scaffolds effectively repaired cartilage defects, with the SF-GMA/LKP10 and SF-GMA/LKP20 groups showing the best results. The scaffolds provided a favorable microenvironment for cell adhesion and proliferation, and the SF-GMA/LKP10 and SF-GMA/LKP20 groups exhibited optimal subchondral bone repair. The study demonstrated that the SF-GMA/LKP composite scaffold is a promising candidate for cartilage regeneration.A self-assembling peptide nanofiber hydrogel (LKP) was developed to mimic the function of TGF-β1 for cartilage regeneration. LKP hydrogel is simple to synthesize and has good biocompatibility and cartilage-promoting ability. However, it has poor mechanical properties and is prone to fragmentation. To address these issues, a series of injectable hydrogel composite scaffolds (SF-GMA/LKP) were prepared by combining LKP with GMA-modified silk fibroin (SF). The SF-GMA/LKP composite scaffolds can induce in-situ filling of cartilage defects and prolong the action time of LKP through interaction between LKP and SF-GMA. The SF-GMA/LKP10 and SF-GMA/LKP20 scaffolds showed the best effect on neocartilage and subchondral bone reconstruction. The composite hydrogel scaffold can be used for high-quality cartilage repair. The SF-GMA/LKP scaffolds were prepared by mixing SF-GMA with LKP peptide nanofiber hydrogels. The scaffolds were characterized using various techniques, including FTIR, SEM, AFM, and rheological analysis. The scaffolds showed good biocompatibility, with no significant toxicity to BMSCs. The SF-GMA/LKP scaffolds promoted the expression of cartilage-related proteins, such as Col-II, Sox9, and ACAN, indicating their potential for cartilage regeneration. In vivo experiments showed that the SF-GMA/LKP scaffolds effectively repaired cartilage defects, with the SF-GMA/LKP10 and SF-GMA/LKP20 groups showing the best results. The scaffolds provided a favorable microenvironment for cell adhesion and proliferation, and the SF-GMA/LKP10 and SF-GMA/LKP20 groups exhibited optimal subchondral bone repair. The study demonstrated that the SF-GMA/LKP composite scaffold is a promising candidate for cartilage regeneration.