The use of nano-sized materials offers exciting new options in technical and medical applications. On the other hand, adverse effects on cells have been reported and may limit their use. In addition to physico-chemical parameters such as contamination with toxic elements, fibrous structure and high surface charge, the generation of radical species was identified as key mechanism for cytotoxic action of nanoparticles. The cytotoxic potential of nanoparticles in the absence of radical generation is less well investigated. This study aims to investigate the size-dependent effect of carboxyl polystyrene particles on cells to identify potential adverse effects of these particles. Particles were characterized in different solutions to assess the influence of the medium on size and surface charge. Viability, membrane integrity, apoptosis, proliferation and generation of oxidative stress were investigated. In addition the intracellular localization of the particles was recorded. 20 nm polystyrene particles induced cellular damage by induction of apoptosis and necrosis. These particles generated radicals to the same degree as larger polystyrene particles. Particles were taken up into endosomes and lysosomes in a size-dependent manner. Protein containing solutions led to increases in particle size, decreased cytotoxicity and reduced cellular uptake. It can be concluded that even in the absence of high surface reactivity and not linked to the generation of radicals nano-sized particles may cause cell damage. The mechanism of this damage includes apoptosis, necrosis and inhibition of proliferation.