Although many reports concluded that polishing of glass ionomers is crucial for smoother surface and limiting the adhesion of cariogenic bacteria, there is no specific surface treatment protocol recommended. A novel material in the same category was released recently claimed to have surface smoothness comparable to resin composite and bacterial adhesion less than other types of glass ionomers. In this study, different polishing systems were tested with three glass ionomers one of them is the novel material to find the most appropriate polishing protocol. Objectives: To evaluate and compare the surface roughness and bacterial adhesion to resin modified glass ionomer, bioactive ionic resin and conventional glass ionomer restorative materials after different polishing protocols in vitro. The materials tested includes resin modified glass ionomer, bioactive ionic resin, and conventional glass ionomer. The polishing protocols were divided into four groups: group 1 = (Mylar matrix strips, Control), group 2 = (one-step, PoGo), group 3 = (two-step, Prisma Gloss) and group 4 = (three-step, Sof-Lex). From each material, eleven cylindrical specimens were prepared for each group according to the manufacturers? instructions. The surface roughness for all specimens was measured using atomic force microscope in tapping mode. the same specimens were subjected to bacterial adhesion testing after being coated with artificial saliva. Data were analyzed with two-way analysis of variance followed by Post hoc multiple comparisons. The highest Ra and S. mutans adhesion values were recorded for all materials in two-step group. The lowest Ra and S. mutans adhesion values were seen in one-step and three step groups. One-step polishing system was more effective and may be preferable for polishing of the three studied glass ionomer-based materials compared to two-step and three-step systems.