β-cyclodextrin and allyl isothiocyanate inclusion complexes (β-CD:AITC) have been proposed for developing fresh fruit and vegetable packaging materials. Therefore, the aim of this research was to develop active materials based on poly(lactic acid) (PLA) loaded with β-CD:AITC and to assess changes in the material properties during the release of AITC to food simulants. PLA films with 0, 5 and 10 wt.% β-CD:AITC were developed by extrusion. Surface properties were determined from contact angle measurements. Films were immersed in water, aqueous and fatty simulants to assess the absorption capacity and the change in the thermal properties. Moreover, the release of AITC in both simulants was evaluated by UV-spectroscopy and kinetic parameters were determined by data modeling. Results showed that a higher concentration of β-CD:AITC increased the absorption of aqueous simulant of films, favoring the plasticization of PLA. However, the incorporation of β-CD:AITC also avoided the swelling of PLA in fatty simulant. These effects and complex relationships between the polymer, inclusion complexes and food simulant explained the non-systematic behavior in the diffusion coefficient. However, the lower partition coefficient and higher percentage of released AITC to the fatty simulant suggested the potential of these materials for high-fat fruit and vegetable active packaging applications.