Hydrogels are widely used in biomedicine and for bioanalytical purposes, normally under wet conditions. For certain applications, processing steps, or process monitoring, hydrogel films are used or treated under ambient conditions, and because they are hygroscopic, it is of interest to investigate how they respond to changes in atmospheric humidity. We have used neutron reflectometry to follow the swelling of thin UV-polymerized hydrogel films in air under different relative humidities (RHs). These polymers were prepared to similar thicknesses on silica and gold substrates, and the chemical similarity between them was verified by infrared spectroscopy. The swelling in response to variations in RH was different for the layers on the two substrate types, reflecting structural changes induced by differences in the UV exposure required to achieve a given polymer thickness, as demonstrated also by differences in the Flory-Huggins interaction parameter, obtained by fitting a Flory-Huggins-type sorption model to the swelling data. Wetting studies show small changes in contact angles with surrounding humidity variations, indicating that structural reorganization at the interface in response to humidity changes is limited.