Background This study investigated the effects of Pro-Argin application on the microhardness of bleached enamel and compared them with those of acidulated phosphate fluoride (APF) and casein phosphopeptide-amorphous calcium phosphate (CPP-ACP) applications. Methods Forty blocks (4x4x4mm) were prepared from the facial surfaces of 10 sound bovine incisors. The enamel surfaces were bleached with 35% hydrogen peroxide three times at 5-day intervals. The specimens were divided into five treatment regimen groups (n=8 each): Group 1, bleaching/no surface treatment (control); Group 2, bleaching/Pro-Argin toothpaste treatment; Group 3, bleaching/Pro-Argin treatment; Group 4, bleaching/APF treatment; and Group 5, bleaching/CPP-ACP treatment. During bleaching treatments, specimens were stored in artificial saliva. Vickers microhardness measurements were performed at baseline and after treatment in all groups. Statistical analyses were performed using the paired t-test, one-way analysis of variance, and post hoc least significant difference tests. Results After bleaching, hardness values of enamel surfaces increased significantly in the treatment groups versus the control group (p<0.05). However, no significant difference was found among treatment groups (p>0.05). Conclusions Through bleaching treatment, Pro-Argin application may have a positive effect on enamel surface hardness, equivalent to that of CPP-ACP or APF.