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CC BY 4.0 · Eur J Dent
DOI: 10.1055/s-0045-1809027
Original Article

The Inhibition of Streptococcus mutans Biofilms following Exposure to Different Chocolate Ingredients

Hadi A. Almoabid   *
1   College of Dentistry, Imam Abdulrahman Bin Faisal University, Dammam, Saudi Arabia
,
Leen Saleh Almutairi    *
1   College of Dentistry, Imam Abdulrahman Bin Faisal University, Dammam, Saudi Arabia
,
Abdul Samad Khan 
2   Department of Restorative Dental Sciences, College of Dentistry, Imam Abdulrahman Bin Faisal University, Dammam, Saudi Arabia
,
Mohammed A. Aljaffary
1   College of Dentistry, Imam Abdulrahman Bin Faisal University, Dammam, Saudi Arabia
,
Rasha AlSheikh
2   Department of Restorative Dental Sciences, College of Dentistry, Imam Abdulrahman Bin Faisal University, Dammam, Saudi Arabia
,
Khalid S. Almulhim
2   Department of Restorative Dental Sciences, College of Dentistry, Imam Abdulrahman Bin Faisal University, Dammam, Saudi Arabia
,
Abdulrahman A. Balhaddad 
2   Department of Restorative Dental Sciences, College of Dentistry, Imam Abdulrahman Bin Faisal University, Dammam, Saudi Arabia
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Abstract

Objectives

This project aimed to investigate the anticariogenic effect of four chocolate ingredients (polyphenol, theobromine, cacao, and flavanol) against Streptococcus mutans biofilms grown in vitro.

Materials and Methods

Stored S. mutans (UA 159) was transferred to Brain Heart Infusion (BHI) broth and incubated in aerobic incubator for 24 hours at 37°C in 5% CO2. Following this, 190 µL of each ingredient concentration (0.78–200 mg/mL) and 10 µL of the culture were added to a 96-well plate and incubated for 24 hours at 37°C in 5% CO2. Then, biofilms were fixed, stained with crystal violet, and analyzed for formation using a spectrophotometer. Control groups included negative control with only S. mutans and sterility control with BHI media.

Statistical Analysis

One-way analysis of variance and Tukey tests analyzed the data.

Results

Flavonoid at the 6.25 to 25 mg/mL concentrations reduced the S. mutans biofilms (p < 0.001) by 5- to 33-fold. Meanwhile, 50 mg/mL concentrations and higher completely eradicated biofilm growth. Similarly, cocoa concentrations ranging between 12.5 and 200 mg/mL revealed massive antibiofilm action from a 22-fold reduction at 12.5 mg/mL to complete biofilm eradication at 200 mg/mL. Polyphenol was the only ingredient showing biofilm inhibition at all concentrations ranging from almost 10-fold reduction to complete biofilm eradication, which were all significant (p < 0.001) compared to the control. Regarding theobromine, 3.125 mg/mL of it significantly increased the growth of S. mutans biofilms. At the concentration of 6.25 mg/mL, theobromine significantly (p < 0.001) inhibited the S. mutans biofilms by 3.35-fold. While at the range of 25 to 200 mg/mL, theobromine resulted in a reduction between 11-fold and complete biofilm eradication.

Conclusion

The findings suggest that flavonoid, cacao, polyphenol, and theobromine may serve as effective adjuncts in preventing dental caries by inhibiting S. mutans biofilm formation.

Keywords

antibacterial - biofilms - caries - chocolate - prevention

Authors' Contributions

All authors made a significant contribution to the work reported, whether that is in the conception, study design, execution, acquisition of data, analysis, and interpretation, or in all these areas; took part in drafting, revising, or critically reviewing the article; gave final approval of the version to be published; have agreed on the journal to which the article has been submitted; and agree to be accountable for all aspects of the work.


Data Availability Statement

The data supporting this study's findings are available from the corresponding author upon reasonable request.


* Both authors contribute equally




Publication History

Article published online:
06 May 2025

© 2025. The Author(s). This is an open access article published by Thieme under the terms of the Creative Commons Attribution License, permitting unrestricted use, distribution, and reproduction so long as the original work is properly cited. (https://creativecommons.org/licenses/by/4.0/)

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