Spontaneous and induced apoptosis of cells obtained from surgical materials after face lift

Abstract

Subcutaneous necrosis after face lift is connected with impaired skin and subcutaneous tissue vascularization as well as with skin cell energy deficiency and activation of the apoptotic signaling pathways. Aim of this work was to measure the level of spontaneous and C2-ceramide-induced apoptosis in cells isolated from surgical materials. Patients. The face subcutaneous tissues samples have been obtained from 52 pa tients (women, 32—64 years old) during plastic surgery for a face lift. Methods. Cell suspensions from surgical samples were prepared by treatment with collagenase followed by centrifugation and incubation in complete medium RPMI 1640 with 10% FCS for 2 h at +37°C, 5% CO₂. Spontaneous apoptosis level was calculated in main cell populations by staining with propidium iodide and flow cytometry. The percentage of induced apoptosis was measured after the culturing of cells with C2-ceramide. Mitochondrial membrane potential was determined by JC-1 staining and flow cytometry. Results. The spontaneous apoptosis levels in all investigated cell populations did notexceed the normal values. C2-ceramide treatment revealed two groups samples with high and low levels of induced apoptosis (18 ± 4% and 37 ± 5% for epithelial cells, and 18 ± 5% and 31 ± 4% for adipocytes). The highest percentage of induced apoptosis was observed for patients over 50 years. The monitoring of patients within 30 days after surgery revealed a prolonged period for wound healing (5.7%), scar infections (1.5%), and scar partial necrosis followed by repeated surgery (1%). Individual case history analysis identified the higher percentage of induced apoptosis and lower mitochondrial membrane potential in these patients. Conclusions. The excessive level of ceramide-induced apoptosis in cells isolated from surgical material has been found toelucidate the high risk of postoperative complications associated with scar necrosis.