Because there is no standard protocol for the removal of resin-based materials that retain semi-rigid trauma splints on teeth, iatrogenic enamel damage caused by various splint removal techniques has remained unknown. The purpose of this study was to evaluate the effect of five different resin removal techniques (H6/H7 scaler, ultrafine diamond bur, ultrasonic scaler, 16-blade tungsten carbide bur, and Sof-Lex disks) on the surface roughness of human enamel. Three-dimensional white-light interferometry, a non-contact profilometry technique, was used to obtain qualitative and quantitative measurements of surface roughness both at baseline and after finishing procedures. Statistical analysis using Friedman test and Wilcoxon signed ranks test showed that the roughest enamel surface was obtained after splint removal with the hand scaler (P < 0.05). Overall, the smoothest enamel surface was obtained after removal of resin with Sof-Lex disks and the 16-blade tungsten carbide bur (P < 0.05).