The Effects of Three Chlorhexidine-Based Mouthwashes on Human Osteoblast-Like SaOS-2 Cells. An In Vitro Study

Abstract

Several decontamination methods for removing biofilm from implant surfaces during surgical peri-implantitis treatment have been reported, including the intraoperative usage of chlorhexidine (CHX)-based antiseptics. There is a lack of information on possible adverse effects on bone healing. The study aimed to examine the impact of three CHX-based mouthwashes on osteoblast-like cells (SaOS-2) in vitro. Cells were cultured for three days in 96-well binding plates. Each well was randomly treated for either 30, 60 or 120 s with 0.05% CHX combined with 0.05% cetylpyridinium chloride (CPC), 0.1% CHX, 0.2% CHX or sterile saline (NaCl) as control. Cell viability, cytotoxicity and apoptosis were assessed at day 0, 3 and 6. Cell viability resulted in being higher in the control group at all time points. At day 0, the CHX 0.2 group showed significantly higher cytotoxicity values compared to CHX 0.1 (30 s), CHX + CPC (30 s, 60 s and 120 s) and control (60 s and 120 s), while no significant differences were identified between CHX + CPC and both CHX 0.1 and NaCl groups. All test mouthwashes were found to induce apoptosis to a lower extent compared to control. Results indicate that 0.2% CHX presented the highest cytotoxic effect. Therefore, its intraoperative use should be carefully considered.

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

“…In a recent in vitro study by our group, a CHX 0.05% + CPC 0.05% mouthwash was revealed to be effective against oral living bacteria after in situ plaque accumulation, showing similar properties as comparted to CHX 0.1% solution [30]. Furthermore, utilizing the same study design of the present work, the authors found the highest cytotoxicity on osteoblast-like cells at day 0 in the CHX 0.2% group, which also presented significantly higher values compared to CHX 0.05% + CPC 0.05% for all the application times [31]. Due to the limited data available on the cytotoxicity on fibroblasts of the former, the current work was conceived as a complementary study in support of our recent investigations [30,31].…”

“…Whereas, at low concentrations, it indirectly promotes cell autolysis through the activation of intracellular latent ribonucleases [29]. Several mouthwash formulations containing both CHX and CPC have been investigated, including solutions with CHX at low concentration, such as CHX 0.05 % + CPC 0.05 % [26,30,31] or CHX 0.03 % + CPC 0.05 % [24,25], but also at higher concentration (e.g., CHX 0.12 % + CPC 0.05 %) [32].…”

Systematic Analysis of Lower Second Molar Periodontal Health after Autogenous Dentine Graft from Wisdom Tooth Extraction

“…One notable departure from the common practice was observed in the study by Kuperschlag et al (2020) [11], where a chlorhexidine mouth rinse was administered before surgery. While there is some evidence supporting its potential to reduce the occurrence of alveolar osteitis, its efficacy in promoting bone healing remains inconclusive [15].…”

Effectiveness of mechanical and chemical decontamination methods for the treatment of dental implant surfaces affected by peri‐implantitis: A systematic review and meta‐analysis

“…It acts through disruption of the cell membranes of bacteria causing cell death (Jenkins et al, 1988 ). While many studies agree that CHX is an effective agent for biofilm reduction on the implant surfaces, there has been recent concern surrounding its cytotoxicity to host cells, influencing the biocompatibility of the implant surface following treatment (Brunello et al, 2021 ; Cai, Li, Wang, Chen, Jiang, Ge, Lei, Huang, 2019 ; Etemadi et al, 2020 ; Ghasemi et al, 2019 ; Kotsakis et al, 2016 ; Ntrouka et al, 2011 ; Tonon et al, 2020 ). For this reason, some authors have suggested eliminating the frequent use of CHX as an agent for biofilm reduction.…”

“…However, the mechanical removal of bacteria from the implant surface with the use of a saline‐soaked cotton pellet is beneficial as a decontamination method (Alhag et al, 2008 ; Kolonidis et al, 2003 ; Persson et al, 1999 ). Supportive authors believe that saline has many benefits as it does not affect the implant surface structure or biocompatibility and is also cheap and readily available (Brunello et al, 2021 ; Kotsakis et al, 2016 ; Monje et al, 2022 ). Although the current meta‐analysis and other in vitro studies show that PBS/saline tends to perform more poorly than other mechanical, laser, or chemical means, in vivo studies where saline was combined with other forms of decontamination showed promising results (Alhag et al, 2008 ; Kolonidis et al, 2003 ; Persson et al, 1999 ).…”