Exposure to elevated concentrations of ambient ultrafine particulate matter has been associated with increased morbidity and mortality in the public. The particle parameters triggering the underlying mechanisms are largely unknown. The aim of this study was to compare biological in vitro-effects of ultrafine model particles (hematite, silicasol) of different composition and different sizes to evaluate the influence of these parameters. Human epithelial (A549) and macrophage (THP-1, Mono Mac 6) cell lines in mono-culture as well as in co-culture were used as cellular models. The uptake of hematite particles into A549 cells was identified by light microscopy and confirmed by transmission electron microscopy. The loss of membrane integrity measured by the lactate dehydrogenase assay as well as the induction of interleukin-6 and interleukin-8 release were affected by the particles in a dose dependent manner. This study demonstrated that particle size and particle composition, respectively, were responsible for the observed biological effects. Furthermore, the co-cultures of epithelial cells (A549) and macrophages (Mono Mac 6 or differentiated THP-1) showed an increased sensitivity to particles concerning the cytokine release in comparison to the mono-cultures of each cell type.