The field of nanobiotechnology holds great promise to develop multiscale fabrication techniques that benefit regenerative medicine. The combination of nano/micro-fabrication techniques with novel biomaterials provides a unique opportunity to create extracellular matrices that mimic native three-dimensional tissue microstructure. This chapter presents advances in nanobiotechnology techniques to develop biomaterial-based matrices and drug delivery systems for regenerative medicine applications. The discussion focuses on systems using hydrogel matrices, nano/micro-fabrication platforms, electrospun nanofibrous matrices, and nano/micro-particles for drug and biomacromolecule delivery. Each technique is discussed from a biomaterials perspective and specific examples are cited to explain their application in tissue repair and regeneration.