In 2013, 26.090 people were killed in road accidents throughout the EU, approximately 6.200 females and 19.800 males. Females account for 51% of the total EU population, but only 24% of all road fatalities. The objective of this research is the analysis of basic road safety parameters related to road users’ gender in the EU countries, by the use of the EU CARE database with disaggregated data on road accidents and of other data sources such as Eurostat. Time-series data on road accident fatalities by gender from 28 EU countries over a period of 10 years (2004–2013) are correlated with basic safety parameters, such as age, road user type and transport mode. Data from the EU Injury Database are also used to identify injury patterns and improve the assessment of injury severity and additional insight into accident causation for road users by gender is offered through the use of in-depth accident data from the EC SafetyNet project Accident Causation System (SNACS). In all EU countries, road fatalities decreased between 2004 and 2013 for both females and males. Besides the trends of that period, the proportion of male road fatalities did not change significantly. Amongst EU countries a slight tendency for male proportions to be higher in south is noted. Additionally, the ratio between male and female fatalities increases with age, reaches a peak between the ages of 30–44 and then falls, which reflects a specific gender development in the travel behaviour of men and women in Europe. As regards the road user type, the proportion of passengers’ or pedestrians’ fatalities is higher for females than the males, while the opposite is true for drivers. The results of the analysis allow for an overall assessment of the gender-differentiated safety level in the European road network, providing thus useful support to decision makers working for the improvement of safety in the European road network.