The paper contains the results of a metallographic examination and nanoindentation test conducted for the medium carbon structural steel with low content of Mn, Si, Cu, Cr, and Ni after its grinding to a depth ranging from 2 μm to 20 μm, at constant cutting speed (peripheral speed) of vs = 25 ms−1 and constant feed rate of vft = 1 m/min. Applied grinding parameters did not cause the surface layer hardening, which could generate an unfavorable stress distribution. The increase in the surface hardness was obtained due to the work hardening effect. Microstructure, phase composition, and chemical composition of the grinded surface layer were examined using an X-ray diffractometer, light microscope, and scanning microscope equipped with X-ray energy-dispersive spectroscopy, respectively. Hardness on the grinded surface and on the cross-section was also determined. It was shown that the grinding of C45 steel causes work hardening of its surface layer without phase transformation. What is more, only grinding to a depth of 20 μm caused the formation of an oxide scale on the work-hardened surface layer. Nanoindentation test on the cross-section, at a short distance from the grinded surface, has shown that ferrite grains were more susceptible to work hardening than pearlite grains due to the creation of an equiaxed cellular microstructure, and that different dislocation substructure was created in the work-hardened surface layer after grinding to different depths.