To assess and compare the enamel surface quality after interproximal enamel reduction (IPR) was performed with different systems and to study the relation between acid penetration depth and enamel surface quality as well as the importance of remineralization. Sixty-five extracted teeth were randomly allocated to five experimental groups: untreated control, manual with New Metal Strips, mechanical with oscillating segment (OS) discs, Safe-Tipped Bur Kit, and the Ortho-Strip, followed by 30 s of polishing with the Softflex system and the Compo-system after treating the tooth with OS discs. Mesial surfaces were demineralized for 24 h and distal surfaces were subjected to interchanging demineralization and remineralization cycles of 24 h each for 18 days. The analysis was carried out by profilometry, scanning electron microscopy, and polarization microscopy. After IPR and polishing, enamel roughness was reduced for all systems tested except for the Essix Safe-Tipped Bur Kit. Subsequent demineralization increased enamel roughness in all groups except controls beyond the original level prior to IPR except for IPR with New Metal Strips or Ortho-Strips and subsequent polishing. Cyclic demineralization and remineralization for 18 days yielded a reduction in acid penetration depth and an increase in surface smoothness, which correlated with each other only for controls and treatment with New Metal Strips or Ortho-Strips. Manual IPR, using New Metal Strips and, even more, the oscillating IPR system Ortho-Strips, yielded smoother interproximal enamel surfaces and less acid penetration depth than the IPR systems with OS discs and the Safe-Tipped Bur Kit after polishing and 18 days of cyclic demineralization and remineralization. Irrespective of the IPR procedure, proper remineralization of IPR-treated surfaces is advisable to reduce caries susceptibility.