The use of a standardized punch of 2 mm diameter for periodontal tissue biopsies seems to be suitable for immunohistochemistry analysis and showed that the GCF may not express all the markers in the same proportion at the corresponding tissue.

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.