Cytotoxicity of modified nonequilibrium plasma with chlorhexidine digluconate on primary cultured human gingival fibroblasts

Abstract

The aim of this study was to investigate the cytotoxicity of modified nonequilibrium plasma with chlorhexidine digluconate (CHX) on human gingival fibroblasts (HGFs), and to evaluate the biosecurity of modified nonequilibrium plasma with 2% CHX as a new method of root canal treatment. Tissue samples taken from human gingiva were primarily cultured and passaged. Cells from passages 3-7 were used. HGFs were treated by modified nonequilibrium plasma with 2% CHX for 0 min (control group), 30 s, 1 min, 1.5 min, 3 min, 5 min, and 10 min, respectively, and then they were incubated for 0, 24, and 48 h. After that, cell counting kit-8 (CCK-8) assay was applied to analyze the cytotoxicity of modified nonequilibrium plasma with 2% CHX on HGFs. There was no significant difference between the 0 h group treated with the modified nonequilibrium plasma for 1 min and the control group (P>0.05). However, there were significant differences between all the other treated groups and the control group (P<0.05). When treated for 1.5 min or shorter, the cell viability was obviously increased; while treated for 3 min or longer, it was obviously reduced. Moreover, when successively cultured for 0, 24, and 48 h, cell viability was decreased at first and then increased in the 3-min-treated and 5-min-treated groups. The modified nonequilibrium plasma with 2% CHX was of no influence on cell viability in 1.5 min treatment, and it could be safely used on root canal treatment.

Cytotoxicity evaluation of chlorhexidine gluconate on human fibroblasts, myoblasts, and osteoblasts

“…In their study, cell viability was significantly reduced when exposed to 2% CHX for 3 minutes or longer. Similar to findings in this study, Chen et al demonstrated that cell viability was reduced in a time-dependent manner, with cells exposed to CHX for 10 minutes having cell survival rates significantly lower than the same cells exposed to CHX for only 3 minutes 21 . Flemingson et al 33 exposed cultured human fibroblasts to different dilutions of commercially available CHX anti-plaque mouthwash products (1%, 2%, 5%, 10%, 20%, and 100%) for 1, 5, and 15 minutes.…”

“…In a study by Cline et al 29 , CHX was shown to affect fibroblast proliferation and impair cell adhesion. Chen et al 21 . exposed human gingival fibroblasts to 2% CHX and used the CCK assay to determine cell survival.…”

“…Prior studies have shown CHX solutions to be cytotoxic to human fibroblasts, osteoblasts, and lymphocytes in a time and dose dependent manner 4 , 8 , 13 , 14 , which may possibly delay wound healing or lead to increased rates of wound dehiscence 15 - 20 . Multiple in vitro studies with CHX has demonstrated its cytotoxicity to fibroblast cells 18 , 20 , 21 . While fibroblasts are a critical cell type in wound healing, myoblasts, and osteoblasts are crucial for skeletal muscle repair and bone healing, respectively 22 - 24 .…”

Use of magnetic nanoparticles as a drug delivery system to improve chlorhexidine antimicrobial activity

“…59 Data presented in this study confirmed that immobilization of CHX on MNPs also decreases the toxic effect of CHX without affecting its killing properties. In effect, magnetic derivatives of CHX may be an interesting alternative for formulations presented by Chen et al 60 Importantly, the employment of nanotechnology-based structures in dentistry provides a way for the introduction of novel therapy approaches based on structures that allow to control drug release and to improve the regenerative processes with subsequent strong antimicrobial activity, even in the presence of salivary proteins. …”

Biochanin A alleviates gingival inflammation and alveolar bone loss in rats with experimental periodontitis

“…The cultured cells from gingiva were tested by immunocytochemical analysis. These cells were tested negative for anti-keratin staining whilst staining positive for vimentin, suggesting these cells to be gingival fibroblasts (23). Cells between sixth and ninth passages were used for the assessment of cell cytotoxicity.…”