The aim of this study was to compare the plaque removal efficacy of a new oscillating/rotating/pulsating toothbrush [Oral-B® Professional Care® 8500 (PC 8500)] with two manual toothbrushes [Oral-B® CrossAction™ Vitalizer (CAV) and Oral-B® Indicator™ (IND), respectively]. The safety of the PC 8500 was also assessed. The study was a single-use, observer-masked, randomised 3 × 3 Latin square crossover design balanced for carryover effects. The enrolled subjects (n = 66) refrained from brushing for 23–25 h before each clinical examination. Plaque scores were recorded before and after brushing with the allocated toothbrush using the Turesky et al. modification of the Quigley and Hein plaque index. The safety was assessed evaluating the soft tissue conditions present after 30 days of the use of the PC 8500. The PC 8500 toothbrush was better in plaque removal efficacy compared with the CAV and IND brushes for full mouth and approximal surfaces (P < 0.01). When marginal surfaces were considered, the PC 8500 was significantly more effective than the IND (P < 0.01). No significant differences were found between PC 8500 and CAV (P > 0.05). The latter was shown to be significantly more effective than the IND at all tooth surfaces (P < 0.01). Safety examinations revealed the onset of only two small gingival abrasions after the 30-day use of the PC 8500. The PC 8500 toothbrush demonstrated to be more effective in plaque control than the CAV and IND in the full mouth and approximal surfaces and similar to the CAV in the marginal surfaces. The PC 8500 was safe to oral tissues in long-term use.