In this study, poly(vinyl-alcohol) (PVA)/chitosan (CS) polymer blend films with different amounts of CS (0, 5, 20 and 35 wt. %) crosslinked by glutaraldehyde (GA) were prepared. The structure and properties of the prepared polymer films were studied by means of dynamic mechanical analysis (DMA), differential scanning calorimetry (DSC), and the time-lag permeation technique. The DMA analysis showed that CS reduces the crystallinity degree of PVA, leading to a higher amount of the amorphous phase contributing to the α relaxation that corresponds to the glass-to-rubber transition. However, the mobility of the amorphous phase can be restricted with crosslinking with 1 wt. % GA. Interaction between the PVA and the CS was confirmed by DCS analysis. Additionally, the influence of the CS and crosslinking on the permeation of nitrogen molecules was investigated. The permeation was examined by the time-lag method. It was found that the addition of CS and GA to PVA improves barrier properties.