This research aims to investigate the impact of advertising signs on road safety. An exhaustive review of international literature was carried out on the effect of advertising signs on driver behaviour and safety. Moreover, a before-and-after statistical analysis with control groups was applied on several road sites with different characteristics in the Athens metropolitan area, in Greece, in order to investigate the correlation between the placement or removal of advertising signs and the related occurrence of road accidents. Road accident data for the ‘before’ and ‘after’ periods on the test sites and the control sites were extracted from the database of the Hellenic Statistical Authority, and the selected ‘before’ and ‘after’ periods vary from 2.5 to 6 years. The statistical analysis shows no statistical correlation between road accidents and advertising signs in none of the nine sites examined, as the confidence intervals of the estimated safety effects are non-significant at 95% confidence level. This can be explained by the fact that, in the examined road sites, drivers are overloaded with information (traffic signs, directions signs, labels of shops, pedestrians and other vehicles, etc.) so that the additional information load from advertising signs may not further distract them.