Purpose: This in vitro study investigated whether aging different restorative materials influences secondary caries development using a short-term in vitro biofilm model, hypothesizing that the antibacterial adhesive employed may lose its effect over time. Materials and Methods: Sixty enamel-dentin blocks were divided into 6 groups with n = 10 per group. The groups were restored with three different restorative materials, of which each sample contained an artificial gap: composite with conventional adhesive (CCA; negative control), composite with an antibacterial adhesive (CAA), and amalgam (A; positive control). Half of the groups were prepared fresh and half of the groups were submitted to an aging protocol consisting of water storage, thermocycling, storage in human saliva, and storage in 0.9% saline solution. All specimens were subjected to an intermittent 1% sucrose biofilm model for 20 days to create artificial caries lesions. Lesion progression in the enamel and dentin next to the different materials was measured as lesion depth (LD) and mineral loss (ML), using transverse wavelength independent microradiography (T-WIM). Regression analysis was used to evaluate the effect of aging on LD and ML per restorative material, corrected for gap size. Results: In the amalgam group, aging led to shallower lesions and less mineral loss. Fresh amalgam samples showed an average lesion depth of 156.65 ± 39.18 µm at wall dentin locations. Aged amalgam samples had an average lesion depth of 73.42 ± 73.50 µm. Fresh CAA samples showed lower average surface mineral loss values (9104 ± 2631 µm•vol%) than did fresh CCA samples (13166 ± 4769 µm•vol%). After aging, this effect was absent, and the average mineral loss in the CAA group was 13382 ± 5586 µm•vol%, while in the CCA group it was 15518 ± 9283 µm•vol%. Conclusion: Aging can influence secondary caries development either positively or negatively depending on the kind of restorative material. Antibacterial adhesives may lose their effectiveness over time. Keywords: adhesives, aging, amalgams, antimicrobial monomers, antibacterial adhesive, bacterial challenge, adhesives, composite restorations, secondary caries