Background, aims. This investigation was designed to evaluate the null hypothesis of no differences in GTR outcomes in intrabony defects at vital and successfully root-canal-treated teeth. Method: 208 consecutive patients with one intrabony defect each were enrolled. Based on tooth vitality, the treated population was divided at baseline into 2 groups: one with 41 non-vital teeth and the other with 167 vital teeth. The 2 groups were similar in terms of patient and defect characteristics. Results: A slight unbalance in terms of depth of the intrabony component was observed in the non-vital group compared to the vital group (6.9 +/-2.1 nm versus 6.2 +/-2.3 mm, p=0.08). All defects were treated with GTR therapy. At 1 year, the non-vital and the vital groups showed a clinical attachment level (CAL) gain of 4.9 +/-2.2 mm and of 4.2 +/-2 mm, respectively. The difference was statistically significant (p=0.03). To correct for the baseline unbalance in defect depth, data were expressed as a % of clinical attachment level gains with respect to the original intrabony depth of the defect. % CAL gains were 72.8 +/- 42.2% and 73 +/- 26.4% for vital and non-vital teeth, respectively: the difference was not statistically significant (p=0.48). Average residual pocket depths were 2.8 +/-1 mm in the vital and 2.8 +/-0.9 mm in the non-vital group. Tooth vitality was assessed at baseline, at 1-year and at follow-up (5.4 +/-2.8 years after surgery): all teeth vital at baseline were still vital at follow-up with the exception of 2 teeth that received endodontic treatment for reconstructive reasons and for caries. At follow-up visit, the difference in CAL with respect to 1-year measurements was -0.9 +/-0.8 mm in the vital group and -0.7 +/-0.8 mm in the non-vital group, indicating stability of the regenerated attachment at the majority of sites. Conclusions: Data from this study demonstrate that root canal treatment does not negatively affect the healing response of deep intrabony defects treated with GTR therapy; furthermore GTR therapy in deep intrabony defects does not negatively influence tooth vitality.