This study aimed to evaluate the effect of different matrix metalloproteinase inhibitors (MMPIs) on the microtensile bond strength (μTBS) and nanoleakage of universal adhesives. One hundred twenty non-carious human molars were prepared and randomly assigned to two groups: Scotchbond Bond Universal (SBU) and Gluma Bond Universal (GBU). The samples in each group were assigned to five subgroups (n=12) based on one control (water) and four MMPIs (Benzalkonium-chloride (BAC), Batimastat (BB94), Chlorhexidine (CHX), and Epigallocatechin-gallate (EGCG)). Each adhesive was applied in self-etch (SE) mode or etch-and-rinse (ER) mode. Dentin/composite sticks were fabricated and subjected to the μTBS test after 24 h or 6 months. At 6 months, MMPIs did not affect the μTBS of the adhesives, regardless of etching mode. Nanoleakage was more pronounced in ER mode than in SE mode for all subgroups. All MMPIs, with the exception of CHX, decreased the nanoleakage of GBU in ER mode.