The purposes of this in vitro study were to evaluate the tubule plugging efficiencies and effects on the surface roughness of dentin of different dentin desensitizing agents (DDAs; Teethmate Desensitizer, Kuraray; Gluma Desensitizer, Kulzer; Clinpro White Varnish, 3M ESPE; Enamelast, Ultradent) and the Er,Cr:YSGG laser (Biolase, Waterlase), both alone and in combination with DDAs, after application and after an erosion-abrasion cycle. For surface roughness examinations, superficial buccal dentin specimens were divided into 10 groups: the control, Teethmate Desensitizer, Gluma Desensitizer, Enamelast, Clipro White Varnish, Er,Cr:YSGG Laser, Teethmate Desensitizer-Laser, Gluma Desensitizer-Laser, and Enamelast-Laser, and Clinpro White Varnish-Laser groups. Profilometric analyses and scanning electron microscopy (SEM) examinations were performed after applications and after a 5-day erosive-abrasive cycle. For the statistical analysis of surface roughness measurements, 2-way analysis of variance (ANOVA), 1-way ANOVA, and Tukey post hoc test were used. Among the treatments, only DDAs alone did not cause increase in surface roughness after application. All of the laser applications increased the surface roughness of dentin, and after the erosion-abrasion cycle, all of the test groups had increased surface roughness. However, SEM images showed that morphological changes were less frequently observed in all of the experimental groups than in the control group. In addition, all of the laser-DDA combinations had stronger tubule occlusion effects than did DDAs alone, even after erosion-abrasion. All of the test treatments showed protective effects on dentin surfaces against the negative effects of erosion-abrasion. The addition of the laser to DDA applications increased tubular plugging efficiencies of DDAs, and the tubule plugs of the combination treatments were resistant to the erosion-abrasion cycle.