Effect of Three Chlorhexidine-Based Mouthwashes on Human Gingival Fibroblasts: An In Vitro Study

Mouthwashes containing chlorhexidine (CHX) are deemed to be associated with dose-dependent side effects, including burning sensation and taste alteration. To overcome these drawbacks, mouthwashes with CHX at lower concentrations with or without adjunctive agents are proposed. The aim of this in vitro study was to investigate the effects of three CHX-based mouthwashes on human gingival fibroblasts (HGFs). After 3 days of cell culture, groups were randomly treated for 30 s, 60 s or 120 s with (a) CHX 0.05% in combination with cetylpyridnium chloride (CPC) 0.05%; (b) CHX 0.1%; (c) CHX 0.2%; or (d) NaCl as control. Cell viability, cytotoxicity and apoptosis were evaluated at 2 h, 3 days and 6 days after the exposure to the different solutions. Similar cell viability values were found among the test groups at all time points. At day 0, higher cytotoxicity was measured in the group treated with CHX 0.2%, in particular after long application time (120 s), while no significant difference was found between CHX + CPC and the control group. All the investigated mouthwashes were well tolerated by HGF cells for the tested application times. The highest cytotoxic effect was observed for CHX 0.2%; therefore, clinicians should consider limiting its usage to carefully selected clinical situations.