AIM: In the present study, we aimed to evaluate the cytosolic and nuclear-mitochondrial expression of pro-apoptotic and anti-apoptotic markers in gingival tissue from patients with severe generalized chronic periodontitis (sGCP). METHODS: Twenty-four patients participated in the study: 15 (54.3 + 8.3 years) with sGCP and nine (38.2 + 5.4 years) with gingivitis alone. Gingival tissue was collected using a 1.5-mm diameter punch and homogenized using a cell disruptor. The supernatants were analyzed for the cytosolic and nuclear-mitochondrial fractions of caspase-3, Bax, Bak, Smac, lamin B, Bad, Bim, survivin, Bcl-xL, Mcl-1, and of the dimers Bcl-2/Bax, Bcl-xL/Bak and Mcl-1/Bak using a multiplex immunoassay. RESULTS: Significantly higher levels of cytosolic Bcl-xL/Bak, nuclear-mitochondrial Mcl-1/Bak, and cytosolic Bcl-xL were observed in gingival tissues from periodontitis patients compared to controls (P = 0.03, 0.03, and 0.05, respectively). The patients with gingivitis presented significantly increased levels of nuclear-mitochondrial Bad, cytosolic and nuclear-mitochondrial Bcl-2/Bax, and cytosolic Bim compared to the patients with periodontitis (P < 0.001, 0.03, 0.05, and 0.04, respectively). CONCLUSION: Significantly higher levels of anti-apoptotic markers, such as Bcl-xL/Bak, Mcl-1/Bak, and Bcl-xL, and lower levels of pro-apoptotic markers Bad and Bim in inflamed tissues indicate an anti-apoptotic trait in patients with sGCP.