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

Abstract

Introduction: Chlorhexidine gluconate (CHX) is widely used as a preoperative surgical skin-preparation solution and intra-wound irrigation agent, with excellent efficacy against wide variety of bacteria. The cytotoxic effect of CHX on local proliferating cells following orthopaedic procedures is largely undescribed. Our aim was to investigate the in vitro effects of CHX on primary fibroblasts, myoblasts, and osteoblasts.Methods: Cells were exposed to CHX dilutions (0%, 0.002%, 0.02%, 0.2%, and 2%) for either a 1, 2, or 3-minute duration. Cell survival was measured using a cytotoxicity assay (Cell Counting Kit-8). Cell migration was measured using a scratch assay: a “scratch” was made in a cell monolayer following CHX exposure, and time to closure of the scratch was measured.Results: All cells exposed to CHX dilutions of ≥ 0.02% for any exposure duration had cell survival rates of less than 6% relative to untreated controls (p < 0.001). Cells exposed to CHX dilution of 0.002% all had significantly lower survival rates relative to control (p < 0.01) with the exception of 1-minute exposure to fibroblasts, which showed 96.4% cell survival (p = 0.78). Scratch defect closure was seen in < 24 hours in all control conditions. However, cells exposed to CHX dilutions ≥ 0.02% had scratch defects that remained open indefinitely.Conclusions: The clinically used concentration of CHX (2%) permanently halts cell migration and significantly reduces survival of in vitro fibroblasts, myoblasts, and osteoblasts. Further in vivo studies are required to examine and optimize CHX safety and efficacy when applied near open incisions or intra-wound application.

Peri-implantitis Update: Risk Indicators, Diagnosis, and Treatment

“…CHX has specific disadvantages because clinically used 2% CHX permanently halts cell migration and significantly reduces fibroblast, myoblast, and osteoblast survival in vitro. 42 Thus, further in vivo studies are required to examine and optimize CHX safety and efficacy.…”

Comparative Study of Antitumor Activity between Lipophilic Bismuth Nanoparticles (BisBAL NPs) and Chlorhexidine on Human Squamous Cell Carcinoma

“…Altogether, these results suggest that CHX kill both tumoral and nontumoral control cells in the same way; altering their cell membrane permeability and damaging their genomic DNA lacking selectivity. Several early studies have been described as the cytotoxicity of CHX on different kinds of cells [13,33,38]. Previously, it was reported that 1% of CHX shows a cytotoxic effect on human gingival fibroblasts (same control cells used in our study) employing also MTT cell viability assays [39].…”

Porphyromonas Gingivalis Load is Balanced by 0.20% Chlorhexidine Gel. A Randomized, Double-Blind, Controlled, Microbiological and Immunohistochemical Human Study

“…In conclusion, the results of this study show that the use of CHX considerably reduces P. gingivalis presence, peri-implant inflammation and improve the clinical outcomes of implant-supported restoration. Different in vitro studies showed how CHX was cytotoxic for fibroblast cells, myoblast, osteoblast, cell viability and migration [62]. However, its in vitro cytotoxicity is higher for concentrations close to 2%.…”