Microstructural internal lengths play an important role on the local and macroscopic mechanical behaviors of steels. In this study, the dislocation density gradients near grain boundaries in a ferritic steel are investigated using SEM/EBSD together with instrumented nanoindentation for undeformed and pre-deformed aluminum-killed steels (Al-k) at 3% and 18% tensile plastic strains. The effect of the distance to grain boundaries on Geometrically Necessary Dislocations (GND) densities is first determined by analyzing orientation gradients from 2D-EBSD maps. Then, nanohardness measurements are performed in the vicinity of grain boundaries. Data analyses show a clear correlation between the spatial gradients of GND density and the ones of nanohardness. Using a mechanistic model, the total dislocation densities are estimated from the measured nanohardness values. From both GND and total dislocation density profiles, the value of an internal length, denoted λ, is estimated from the analysis of dislocation density gradients near grain boundaries. At the end, the capabilities of 2D-EBSD and nanoindentation methods to assess this value are discussed.