The purpose of the present study was to histologically assess the effect of enamel matrix derivative on periodontal regeneration in one human experimental defect. Experimental surgery was performed in a healthy male volunteer to create a buccal dehiscence defect in a mandibular incisor. Following bone removal and conditioning of the exposed root surface, enamel matrix derivative was applied onto the root surface. The flaps were then replaced and sutured. Clinical healing was uneventful. After 4 months, the experimental tooth together with the surrounding soft and hard tissues were removed surgically for histological evaluation. The microscopic examination revealed formation of a new acellular extrinsic fibre cementum, which was firmly attached to the underlying dentin. A new periodontal ligament with inserting and functionally-oriented collagen fibres and an associated alveolar bone was also present. The new cementum covered 73% of the original defect. Regain of bone was 65% of the presurgical bone height. It was concluded that adjunctive use of enamel matrix derivative could provide a regenerative technology with a potential for true periodontal regeneration.