In Vitro Evaluation of Antibacterial and Antibiofilm Activity of Different Chlorhexidine-Containing Mouthwash Formulations against <i>Streptococcus mutans</i>
Daily use of mouthwash is generally recommended to control dental plaque development and limit further adhesion of oral bacteria. To this purpose, Chlorhexidine di-gluconate (CHG)-containing products are still the most used antiseptic in oral health. The aim of this study was to compare the antimicr...
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MDPI AG
2023-06-01
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| Series: | Applied Sciences |
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| Online Access: | https://www.mdpi.com/2076-3417/13/13/7531 |
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| author | Francesca Mensitieri Mario Caggiano Giulia Gaudino Bruno Charlier Albino Coglianese Alessandra Amato Federica Di Spirito Massimo Amato Fabrizio Dal Piaz Viviana Izzo |
| author_facet | Francesca Mensitieri Mario Caggiano Giulia Gaudino Bruno Charlier Albino Coglianese Alessandra Amato Federica Di Spirito Massimo Amato Fabrizio Dal Piaz Viviana Izzo |
| author_sort | Francesca Mensitieri |
| collection | DOAJ |
| description | Daily use of mouthwash is generally recommended to control dental plaque development and limit further adhesion of oral bacteria. To this purpose, Chlorhexidine di-gluconate (CHG)-containing products are still the most used antiseptic in oral health. The aim of this study was to compare the antimicrobial and antibiofilm activity of three different mouthwash commercial formulations containing CHG at the same concentration (0.2%) on the main etiological agent of dental plaque, <i>Streptococcus mutans</i> (<i>S. mutans</i>). Liquid Chromatography coupled with tandem mass spectrometry (LC-MS/MS) was used to confirm the CHG concentration in the commercial formulations. The Minimal Inhibitory Concentration (MIC) and Minimal Bactericidal concentration (MBC) were measured to evaluate the antimicrobial activity of mouthwashes on planktonic cells. The biofilm prevention concentration (BPC) and CHG formulation activity towards mature biofilm were investigated. Both total cell titer and viability in <i>S. mutans</i> pregrown biofilms were evaluated after treatment with formulations at different time points and concentrations, using Crystal Violet (CV) and tetrazolium dye assay (MTT). The Shapiro–Wilk and Levene tests were used to evaluate the normality and overall homogeneity of the datasets, respectively. The chi-square test, Fischer’s exact test and Student’s <i>t</i>-test were used to evaluate the inhibitory capacity of the commercial formulations in CV and MTT assays. Results showed that even though only slight differences in MIC and MBC values were found on planktonic bacteria, the antibiofilm effect of the different formulations exhibited wide variation when mature biofilms were treated. In addition, results were discussed based on the different compositions of the three formulations, suggesting that herbal extracts, when present in mouthwash formulations, may not always have a beneficial effect, especially when mature biofilms are concerned. |
| first_indexed | 2024-03-11T01:46:58Z |
| format | Article |
| id | doaj.art-421583fd72c94fb396256b5d4a2c5782 |
| institution | Directory Open Access Journal |
| issn | 2076-3417 |
| language | English |
| last_indexed | 2024-03-11T01:46:58Z |
| publishDate | 2023-06-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Applied Sciences |
| spelling | doaj.art-421583fd72c94fb396256b5d4a2c57822023-11-18T16:07:46ZengMDPI AGApplied Sciences2076-34172023-06-011313753110.3390/app13137531In Vitro Evaluation of Antibacterial and Antibiofilm Activity of Different Chlorhexidine-Containing Mouthwash Formulations against <i>Streptococcus mutans</i>Francesca Mensitieri0Mario Caggiano1Giulia Gaudino2Bruno Charlier3Albino Coglianese4Alessandra Amato5Federica Di Spirito6Massimo Amato7Fabrizio Dal Piaz8Viviana Izzo9Department of Medicine, Surgery and Dentistry “Scuola Medica Salernitana”, University of Salerno, 84084 Baronissi, ItalyDepartment of Medicine, Surgery and Dentistry “Scuola Medica Salernitana”, University of Salerno, 84084 Baronissi, ItalyDepartment of Medicine, Surgery and Dentistry “Scuola Medica Salernitana”, University of Salerno, 84084 Baronissi, ItalyDepartment of Medicine, Surgery and Dentistry “Scuola Medica Salernitana”, University of Salerno, 84084 Baronissi, ItalyDepartment of Medicine, Surgery and Dentistry “Scuola Medica Salernitana”, University of Salerno, 84084 Baronissi, ItalyDepartment of Medicine, Surgery and Dentistry “Scuola Medica Salernitana”, University of Salerno, 84084 Baronissi, ItalyDepartment of Medicine, Surgery and Dentistry “Scuola Medica Salernitana”, University of Salerno, 84084 Baronissi, ItalyDepartment of Medicine, Surgery and Dentistry “Scuola Medica Salernitana”, University of Salerno, 84084 Baronissi, ItalyDepartment of Medicine, Surgery and Dentistry “Scuola Medica Salernitana”, University of Salerno, 84084 Baronissi, ItalyDepartment of Medicine, Surgery and Dentistry “Scuola Medica Salernitana”, University of Salerno, 84084 Baronissi, ItalyDaily use of mouthwash is generally recommended to control dental plaque development and limit further adhesion of oral bacteria. To this purpose, Chlorhexidine di-gluconate (CHG)-containing products are still the most used antiseptic in oral health. The aim of this study was to compare the antimicrobial and antibiofilm activity of three different mouthwash commercial formulations containing CHG at the same concentration (0.2%) on the main etiological agent of dental plaque, <i>Streptococcus mutans</i> (<i>S. mutans</i>). Liquid Chromatography coupled with tandem mass spectrometry (LC-MS/MS) was used to confirm the CHG concentration in the commercial formulations. The Minimal Inhibitory Concentration (MIC) and Minimal Bactericidal concentration (MBC) were measured to evaluate the antimicrobial activity of mouthwashes on planktonic cells. The biofilm prevention concentration (BPC) and CHG formulation activity towards mature biofilm were investigated. Both total cell titer and viability in <i>S. mutans</i> pregrown biofilms were evaluated after treatment with formulations at different time points and concentrations, using Crystal Violet (CV) and tetrazolium dye assay (MTT). The Shapiro–Wilk and Levene tests were used to evaluate the normality and overall homogeneity of the datasets, respectively. The chi-square test, Fischer’s exact test and Student’s <i>t</i>-test were used to evaluate the inhibitory capacity of the commercial formulations in CV and MTT assays. Results showed that even though only slight differences in MIC and MBC values were found on planktonic bacteria, the antibiofilm effect of the different formulations exhibited wide variation when mature biofilms were treated. In addition, results were discussed based on the different compositions of the three formulations, suggesting that herbal extracts, when present in mouthwash formulations, may not always have a beneficial effect, especially when mature biofilms are concerned.https://www.mdpi.com/2076-3417/13/13/7531chlorhexidine<i>S. mutans</i>mouthwashantibacterial activityantibiofilm activityoral microbiome |
| spellingShingle | Francesca Mensitieri Mario Caggiano Giulia Gaudino Bruno Charlier Albino Coglianese Alessandra Amato Federica Di Spirito Massimo Amato Fabrizio Dal Piaz Viviana Izzo In Vitro Evaluation of Antibacterial and Antibiofilm Activity of Different Chlorhexidine-Containing Mouthwash Formulations against <i>Streptococcus mutans</i> Applied Sciences chlorhexidine <i>S. mutans</i> mouthwash antibacterial activity antibiofilm activity oral microbiome |
| title | In Vitro Evaluation of Antibacterial and Antibiofilm Activity of Different Chlorhexidine-Containing Mouthwash Formulations against <i>Streptococcus mutans</i> |
| title_full | In Vitro Evaluation of Antibacterial and Antibiofilm Activity of Different Chlorhexidine-Containing Mouthwash Formulations against <i>Streptococcus mutans</i> |
| title_fullStr | In Vitro Evaluation of Antibacterial and Antibiofilm Activity of Different Chlorhexidine-Containing Mouthwash Formulations against <i>Streptococcus mutans</i> |
| title_full_unstemmed | In Vitro Evaluation of Antibacterial and Antibiofilm Activity of Different Chlorhexidine-Containing Mouthwash Formulations against <i>Streptococcus mutans</i> |
| title_short | In Vitro Evaluation of Antibacterial and Antibiofilm Activity of Different Chlorhexidine-Containing Mouthwash Formulations against <i>Streptococcus mutans</i> |
| title_sort | in vitro evaluation of antibacterial and antibiofilm activity of different chlorhexidine containing mouthwash formulations against i streptococcus mutans i |
| topic | chlorhexidine <i>S. mutans</i> mouthwash antibacterial activity antibiofilm activity oral microbiome |
| url | https://www.mdpi.com/2076-3417/13/13/7531 |
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