Objectives Chronic periodontitis is a result of an unsuccessful resolution of an inflammation. Normally, specialised pro-resolution mediators (SPMs), lipoxins and resolvins, help to stop and resolve the inflammation. Endogenous synthesis of lipoxins and resolvins from fatty acids (PUFAs) can be induced by aspirin’s action on cyclooxygenase-2. The aim of this work is to assess the use of granulation tissue enriched with aspirin and PUFAs in the treatment of chronic periodontitis. Methods In 24 rabbits, a ligature-induced periodontitis model was used to create 47 periodontal defects. Six weeks after the induction, the surgery was performed to extract the granulation tissue (GT) from the defect. In group 1, GT was soaked with aspirin and PUFAs and replanted back. In group 2, soaking in saline served as placebo before giving it back into the defect. In group 3, GT was not returned. Animals were sacrificed after 2, 6 and 12 weeks and samples were harvested for histological and molecular biological analysis. Primary observed parameters were probing pocket depth (PPD) and clinical attachment level (CAL) measured before the surgery and after the necropsy. Results Six weeks after the surgery, PPD and CAL were significantly lower in group 1, treated with aspirin and PUFAs, compared with groups 2 and 3 (p<0,05). Differences of PPD and CAL in other end-points did not reach statistical significance. Also, in epithelialisation, reactive cellulisation, inflammatory infiltrate and size of bony defect, no significant differences were found, and all the defects healed. Conclusion Trend of an earlier healing can be seen when treating a periodontal defect with GT soaked with aspirin and PUFAs. However, 12 weeks after the intervention, no results can be found between the three groups anymore. Yet, we can say that enriched GT may be successfully returned into the periodontal defect with similar treatment outcomes as standard removal of GT in periodontal surgery techniques. It needs to be determined, whether GT only served as a carrier, or it was directly affected by aspirin and PUFAs and changed its behaviour from pro-inflammatory to pro-resolution, via enhanced lipoxin and resolvin synthetic pathways. Our results may be confirmed and further elucidated in a following clinical study on patients.