OBJECTIVES: The aim of this study was to evaluate the in vitro effects of chlorhexidine mouthrinse solution (0.12%), on the morphology and cell viability of three different cell populations of human periodontal tissues, including osteoblastic cells, periodontal ligament cells (PDL), and gingival fibroblasts (GF), in a time-dependent mode.
MATERIALS AND METHODS: PDL and GF cell cultures were derived from maxillary premolars, and osteoblastic cells from human osteosarcoma (MG-63 cell-line). Cells were cultured in Dulbecco's modified eagle's medium (DMEM), supplemented with 10% fetal bovine serum (FBS) and antibiotics, followed by treatment with CHX solution in concentrations of 0,12% (v/v). Cell viability was performed using Trypan Blue Exclusion Assay. Morphological changes were evaluated by high-definition phase contrast microscopy, at selected time intervals. Statistical analysis was performed using Poisson regression analysis.
RESULTS: The effects of CHX on the cells were immediate and profound. The intensity of the morphological changes was gradually increasing, analogously to exposure time. The most characteristic changes observed, included gradual shrinking of the cells, widening of the intercellular spaces, gradual loss of cytoplasmic organelles and appearance of vacuoles. The intensity of the changes on PDL and GF cells was greater than that on the osteoblastic cells. After 24 hours, all three types of cell-cultures showed signs of complete cell lysis. Cell viability of all cell types was gradually reduced in a time dependent mode.
CONCLUSIONS: The 0.12% CHX solution seems to have an immediate cytotoxic effect on human periodontal cells, affecting the cell structure and viability in a time-dependent manner.