Cytotoxicity of chlorhexidine-hydrogen peroxide combination in different concentrations on cultured human periodontal ligament fibroblasts

• Main Authors: Hosein Mirhadi, Mohammad Reza Azar, Abbas Abbaszadegan, Bita Geramizadeh, Shima Torabi, Marjan Rahsaz
• Format: Article
• Language: English
• Published: Wolters Kluwer Medknow Publications 2014-01-01
• Series: Dental Research Journal
• Subjects: Chlorhexidine; cytotoxicity; hydrogen peroxide; root canal irrigation solution
• Online Access: http://www.drjjournal.net/article.asp?issn=1735-3327;year=2014;volume=11;issue=6;spage=645;epage=648;aulast=Mirhadi
• ISSN: 1735-3327; 2008-0255

Background: A strong antimicrobial synergism between chlorhexidine (CHX) and hydrogen peroxide (H 2 O 2 ) has been reported, but there is not enough data on the cytotoxicity of this combination. The primary aim of this study was to evaluate the cytotoxicity of CHX-H 2 O 2 combination in different concentrations and secondary aim is to assess the influence of H 2 O 2 on cytotoxicity of CHX on cultured human periodontal ligament (PDL) fibroblasts. Materials and Methods: The PDL cells were cultured from healthy human third molar teeth and were exposed to six prepared solutions (0.2% and 2% CHX separately and in combination with 1% and 3% H 2 O 2 ). The MTT assay was applied to assess their effects on the viability of the PDL cells. Two-way analysis of variance approach and subgroup analysis was performed to evaluate the differences in mean cell viability values. A level of P < 0.05 was considered as statistically significant. Results: All tested solutions were toxic to PDL cells. There was a significant interaction effect between CHX and H 2 O 2. The 2% CHX combined with 3% H 2 O 2 was the most and 0.2% CHX was the least cytotoxic solutions. The 2% CHX was significantly more toxic than 0.2% CHX and H 2 O 2 combinations. The cytotoxicity of 0.2% CHX and H 2 O 2 combinations did not significantly rise by increasing the concentration of H 2 O 2 from 1% to 3%. Conclusion: H 2 O 2 affected the cytotoxicity of CHX in a variable concentration-dependent manner. Based on the results of this study, it can be concluded that 2% CHX alone and in combination with either 1 or 3% H 2 O 2 are significantly more toxic than 0.2% CHX alone and in combination with 1 and 3% H 2 O 2 . Therefore, to benefit from the synergistic antimicrobial effect between CHX and H 2 O 2 , with a minimal cytotoxicity, it is recommended to use 0.2% concentration of CHX combined with 3% H 2 O 2 .