The bleaching efficacy of common bleaching agents and deionized water treated with non-thermal atmospheric pressure plasma in the pulp chamber for nonvital tooth bleaching was evaluated. A total of 120 extracted human maxillary first incisors were stained using human blood. Teeth were randomly divided into eight groups (n = 15). In the first four groups, teeth were bleached using 35% hydrogen peroxide gel, 37% carbamide peroxide gel, 2:1 (w/v) sodium perborate paste, and deionized water for 30 min. In the remaining groups, bleaching agents were treated with non-thermal atmospheric plasma for 5 min inside the pulp chamber. Overall color changes (∆E) were determined using Commission Internationale de L’Eclairage Lab Colour System. The plasma-assisted tooth bleaching has not increased tooth temperature beyond 37°C. Bleaching efficacies of bleaching agents were significantly improved when treated with non-thermal atmospheric plasma compared to their application (P < 0.05). A remarkable bleaching effect was obtained when bleaching agents were substituted with water and when treated with non-thermal atmospheric plasma. Non-thermal atmospheric plasma treatment could be a novel tool for activation of bleaching agents in the pulp chamber for nonvital tooth bleaching procedure. Moreover, water could be used as a novel bleaching agent when treated with the non-thermal atmospheric plasma to eliminate possible risks which might arise from peroxide-containing agents.