Objective To systematically review the available histologic evidence on periodontal regeneration in class II and III furcations in animals and humans. Materials and methods A protocol including all aspects of a systematic review methodology was developed including definition of the focused question, defined search strategy, study inclusion criteria, determination of outcome measures, screening methods, data extraction and analysis, and data synthesis. The focused question was defined as follows: BWhat is the regenerative effect obtained by using or not several biomaterials as adjuncts to open flap surgery in the treatment of periodontal furcation defects as evaluated in animal and human histological studies?Ŝ earch strategy Using the MEDLINE database, the literature was searched for articles published up to and including September 2018: combinations of several search terms were applied to identify appropriate studies. Reference lists of review articles and of the included articles in the present review were screened. A hand search of the most important dental journals was also performed. Criteria for study selection and inclusion Only articles published in English describing animal and human histological studies evaluating the effect of surgical treatment, with or without the adjunctive use of potentially regenerative materials (i.e., barrier membranes, grafting materials, growth factors/proteins, and combinations thereof) for the treatment of periodontal furcation defects were considered. Only studies reporting a minimum of 8 weeks healing following reconstructive surgery were included. The primary outcome variable was formation of periodontal supporting tissues [e.g., periodontal ligament, root cementum, and alveolar bone, given as linear measurements (in mm) or as a percentage of the instrumented root length (%)] following surgical treatment with or without regenerative materials, as determined histologically/histomorphometrically. Healing type and defect resolution (i.e., complete regeneration, long junctional epithelium, connective tissue attachment, connective tissue adhesion, or osseous repair) were also recorded. Results In animals, periodontal regeneration was reported in class II and III defects with open flap debridement alone or combined with various types of bone grafts/bone substitues, biological factors, guided tissue regeneration, and different combinations thereof. The use of biological factors and combination approaches provided the best outcomes for class II defects whereas in class III defects, the combination approaches seem to offer the highest regenerative outcomes. In human class II furcations, the best outcomes were obtained with DFDBA combined with rhPDGF-BB and with GTR. In class III furcations, evidence from two case reports indicated very limited to no periodontal regeneration.