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With the objective of correlating the anatomical aspects of the palato-gingival groove with its etiology, diagnosis, and alternative treatments, 13 permanent maxillary incisors with palato-gingival grooves were selected from a large sample and subjected to macroscopic and microscopic analysis of groove morphology. The palato-gingival groove occurred most frequently on the lingual aspect of the lateral incisor (11 of 13), and its coronal and radicular extensions were on the disto-lingual surface of the incisors (7 of 13 and 6 of 13, respectively). Deformation of the contour of the pulp cavity was noted subjacent to the groove (9 of 13), along with diminished enamel and dentin thickness (11 of 13 and 13 of 13, respectively) and an increase in cement (12 of 13). The groove was observed extending to the apical third in nine specimens, and a direct communication between the pulp and periodontium was observed in only one case. From these examinations it is concluded that the palato-gingival groove can be clinically diagnosed, preventing subsequent problems; however microscopic analysis of the affected tooth is necessary to allow precise evaluation of the groove's extension and damage to the dental structure.