The aim of this report is to contribute to a better understanding of the radiographic, clinical and anatomic findings in maxillary second premolars. This paper reports the endodontic treatment of two cases of three-rooted three-canal maxillary second premolars in different patients, and two sound maxillary second premolars also with three canals and three independent roots in a sibling of one of the patients. Although the presence of maxillary second premolars with one or two canals and one root is much more common, other anatomic conditions can be found. A correct clinical and radiographic diagnosis based on knowledge of root canal anatomy and critical interpretation of radiographs is necessary for a safer and successful endodontic treatment of these teeth.