Graphene is a two-dimensional carbon nanomaterial extensively explored due to its excellent mechanical, thermal, chemical, optical, electrical, electronic, and medical properties. Due to its promising characteristics, they are widely used in composites, batteries, flexible transparent screens, supercapacitors, electronic devices, sensors, advanced biomaterials, etc. Graphene and its derivatives are explored in the biomedical field as an excellent advanced biomaterial. Considering the range of biocompatibility, large surface area, chemical purity, ease of functionalization, and good drug loading efficiency, graphene-based nanomaterials are employed in various biomedical applications. This chapter highlights the major biomedical applications of graphene-based nanomaterials including biosensing, bioimaging, drug delivery, tissue engineering, phototherapy, and theranostics.