This in vitro study investigated whether restoration materials and adhesives influence secondary caries formation in gaps using a short-term in vitro biofilm model. Sixty enamel-dentin blocks were restored with 6 different restoration materials with or without adhesives (n = 10 per group) with a gap: 1) Clearfil AP-X composite, 2) Clearfil AP-X composite + SE Bond, 3) Clearfil AP-X composite + ProtectBond, 4) Filtek Silorane composite, 5) Filtek Silorane composite + Silorane System adhesive, or 6) Tytin amalgam. Specimens were subjected to an intermittent 1% sucrose biofilm model for 20 days to create artificial caries lesions. Lesion progression in the enamel-dentin next to the different materials was measured in lesion depth (LD) and mineral loss (ML) using transversal wavelength independent microradiography (T-WIM). A regression analysis was used to compare the LD and ML of the different restoration materials at 4 measurement locations: 1 location at the surface of the enamel, 1 location at the wall of the enamel, and 2 locations at the wall of the dentin. A statistically significant effect of AP-X composite with Protect Bond was found for LD and ML at the WallDentin1 location, leading to less advanced wall lesions. An additional finding was that gap size was also statistically significant at the 2 wall locations in dentin, leading to increasing lesion progression with wider gaps. In conclusion, adhesives can influence wall lesion development in gaps. Protect Bond showed significantly less caries progression compared to bare restoration materials or other adhesives in this short-term in vitro biofilm model.