Purpose: To compare the removal of dental biofilm with soft and medium filament toothbrushes. Materials and Methods: Twenty-five patients refrained from performing mechanical or chemical plaque control for 96 hours prior to the experiment in order to allow dental biofilm accumulation on the tooth surfaces. After the period of dental biofilm accumulation, quadrants were randomised and participants brushed different quadrants with soft and medium toothbrushes, with or without dentifrice. After plaque disclosure, the Quigley-Hein (Turesky) plaque index was assessed by a blinded examiner before and after toothbrushing. In addition, photographs before and after brushing were taken for evaluation of gingival abrasion. Results: The results revealed that medium toothbrushes removed high amounts of dental biofilm compared to soft toothbrushes (P 0.05). Considering gingival abrasion, intergroup analysis demonstrated a statistically significant difference on the cervical regions, with a higher proportion of abrasions in the medium toothbrush group (P 0.01). Conclusion: Medium toothbrushes have a greater ability to remove biofilm and cause gingival abrasion than do soft toothbrushes. Keywords: toothbrushing, dental plaque, gingival recession