Silk, recognized as a premium textile material, has a history dating back to 2000–2400 BC. Derived primarily from Bombyx mori larvae, silk fibroin exhibits exceptional biocompatibility, biodegradability, and mechanical properties, making it ideal for biomedical applications. This chapter explores the structural and functional properties of silk fibroin, its processing techniques, and its versatile applications in tissue engineering, drug delivery, and regenerative medicine. Various preparation techniques, formats, and current clinical uses and prospects of silk fibroin-based materials are discussed in brief, highlighting its potential to revolutionize biomedical engineering.