Aim To investigate tooth survival and clinical long-term outcomes up to 26 years following guided tissue regeneration (GTR) therapy in deep intra-bony defects. Methods Patients from three prospective clinical split-mouth studies, which investigated the outcomes of GTR therapy, were re-evaluated 21-26 years after surgery independent of the membrane type used, and tooth survival was assessed according to several site-specific and patient-related factors. Results About 50 patients contributing 102 defects were available for this long-term follow-up. After up to 26 years (median 23.3 years), 52.9% of the teeth were still in situ. The median survival of the extracted teeth was 13.8 years. Patients with diabetes mellitus and/or smoking history lost significantly more teeth in the long term. Compared to the 1-year situation, there was no new median CAL loss after up to 26 years in the teeth which were still in situ. Conclusions Within the limitations of this study, our data show that more than 50% of the initially seriously diseased teeth were still in situ up to 26 years following GTR therapy despite an overall limited adherence to SPT. In the majority of these teeth, the CAL gain 1 year after GTR could be maintained over this long period.