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In this work, we present a short summary of the synthesis and characterization of superparamagnetic iron oxide nanoparticles and their behavior in vitro and in vivo. Therefore, we have used various characterization techniques to deduce the physical particle size as well as magnetic properties. It is shown that the particle properties were significantly improved by a thermochemical treatment and dialysis, obtaining weakly interacting particles with a clear blocking temperature. We also present the interaction of polyvinyl alcohol and vinyl alcohol/vinyl amine copolymer-coated SPIONs with HELA cells. It is shown that the uptake increased significantly in the presence of a magnetic field and that surface functional groups had an impact on particle uptake and metabolic activity. Furthermore, the influences of the varied parameters (polymer type and therefore surface charge, cell medium, and serum) on the agglomeration rate and the cell uptake are presented and discussed. Finally, we briefly describe the intraarticular application of SPIONs in sheep, their uptake by synovial membrane, and their systemic distribution and elimination.