Nuclear Magnetic Resonance (NMR) based diffusion methods open new perspectives for nanomedicine characterization and their bioenvironment interaction understanding. This review summarizes the theoretical background of diffusion phenomena. Self-diffusion and mutual diffusion coefficient notions are featured. Principles, advantages, drawbacks, and key challenges of NMR diffusometry spectroscopic and imaging methods are presented. This review article also gives an overview of representative applicative works to the nanomedicine field that can contribute to elucidate important issues. Examples of in vitro characterizations such as identification of formulated species, process monitoring, drug release follow-up, 2 / 29 nanomedicine interactions with biological barriers are presented as well as possible transpositions for studying in vivo nanomedicine fate.