Silk fibroin (SF) is a natural fibrous protein with great potential in tissue engineering. This review summarizes the latest developments in SF-based scaffolds for tissue engineering, focusing on their structure, processing methods, and applications in skin, bone, cartilage, blood vessels, ligaments, tendons, and nerves. SF scaffolds mimic the extracellular matrix, providing a physical structure for cell interaction and tissue regeneration. The primary structure of SF includes an outer sericin layer and two inner fibroin layers, while the secondary structure consists of silk I, II, and III, which determine the material's properties. SF is extracted from silk cocoons through degumming, dissolution, and various processing techniques such as electrospinning, freeze-drying, solvent casting, gas foaming, and 3D printing. These methods allow the fabrication of scaffolds with different morphologies and porosities, which are crucial for tissue regeneration. SF scaffolds have been applied in various tissue engineering applications, including skin regeneration, bone repair, cartilage regeneration, blood vessel tissue engineering, ligament and tendon regeneration, and nerve tissue regeneration. SF scaffolds offer advantages such as biocompatibility, biodegradability, and the ability to support cell adhesion and proliferation. However, challenges remain in balancing biocompatibility and antibacterial properties, as well as ensuring long-term effectiveness in clinical applications. Overall, SF-based scaffolds show great promise in tissue engineering due to their unique properties and potential for customization.Silk fibroin (SF) is a natural fibrous protein with great potential in tissue engineering. This review summarizes the latest developments in SF-based scaffolds for tissue engineering, focusing on their structure, processing methods, and applications in skin, bone, cartilage, blood vessels, ligaments, tendons, and nerves. SF scaffolds mimic the extracellular matrix, providing a physical structure for cell interaction and tissue regeneration. The primary structure of SF includes an outer sericin layer and two inner fibroin layers, while the secondary structure consists of silk I, II, and III, which determine the material's properties. SF is extracted from silk cocoons through degumming, dissolution, and various processing techniques such as electrospinning, freeze-drying, solvent casting, gas foaming, and 3D printing. These methods allow the fabrication of scaffolds with different morphologies and porosities, which are crucial for tissue regeneration. SF scaffolds have been applied in various tissue engineering applications, including skin regeneration, bone repair, cartilage regeneration, blood vessel tissue engineering, ligament and tendon regeneration, and nerve tissue regeneration. SF scaffolds offer advantages such as biocompatibility, biodegradability, and the ability to support cell adhesion and proliferation. However, challenges remain in balancing biocompatibility and antibacterial properties, as well as ensuring long-term effectiveness in clinical applications. Overall, SF-based scaffolds show great promise in tissue engineering due to their unique properties and potential for customization.