Anti-Bacterial Effect of Cannabidiol against the Cariogenic Streptococcus mutans Bacterium: An In Vitro Study
Abstract
:1. Introduction
2. Results
2.1. CBD Inhibits Planktonic Growth and Biofilm Formation of S. mutans
2.2. CBD Treatment Prevents the Drop in pH Caused by S. mutans
2.3. CBD Reduces the Metabolic Activity and Prevents the Recovery of Preformed Biofilm of S. mutans
2.4. Precoating of Surfaces with CBD Alters Bacterial Growth and Biofilm Formation
2.5. CBD Decreases the Number of Live Bacteria and the Amount of EPS Produced in S. mutans Biofilms
2.6. CBD Increases the Membrane Potential of S. mutans
3. Discussion
4. Materials and Methods
4.1. Materials
4.2. Bacteria Source and Cultivation
4.3. Determination of Minimal Inhibitory Concentration (MIC)
4.4. Determination of Minimal Biofilm Inhibitory Concentration (MBIC)
4.5. pH Measurements of Planktonic Growing Bacteria
4.6. Effect of CBD on Preformed Biofilms
4.7. Biofilm Recovery after Drug Removal
4.8. Effect of CBD Precoating of Culture Plate Surfaces on Biofilm Formation
4.9. Biofilm Analysis by Spinning Disk Confocal Microscopy (SDCM)
4.10. Membrane Potential Assay
4.11. Statistical Analysis
5. Conclusions
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
- Pisanti, S.; Malfitano, A.M.; Ciaglia, E.; Lamberti, A.; Ranieri, R.; Cuomo, G.; Abate, M.; Faggiana, G.; Proto, M.C.; Fiore, D.; et al. Cannabidiol: State of the art and new challenges for therapeutic applications. Pharmacol. Ther. 2017, 175, 133–150. [Google Scholar] [CrossRef] [PubMed]
- Odieka, A.E.; Obuzor, G.U.; Oyedeji, O.O.; Gondwe, M.; Hosu, Y.S.; Oyedeji, A.O. The medicinal natural products of Cannabis sativa Linn.: A review. Molecules 2022, 27, 1689. [Google Scholar] [CrossRef] [PubMed]
- Hanuš, L.O.; Meyer, S.M.; Muñoz, E.; Taglialatela-Scafati, O.; Appendino, G. Phytocannabinoids: A unified critical inventory. Nat. Prod. Rep. 2016, 33, 1357–1392. [Google Scholar] [CrossRef] [Green Version]
- Mechoulam, R.; Hanuš, L.O.; Pertwee, R.; Howlett, A.C. Early phytocannabinoid chemistry to endocannabinoids and beyond. Nat. Rev. Neurosci. 2014, 15, 757–764. [Google Scholar] [CrossRef] [PubMed]
- Cristino, L.; Bisogno, T.; Di Marzo, V. Cannabinoids and the expanded endocannabinoid system in neurological disorders. Nat. Rev. Neurol. 2020, 16, 9–29. [Google Scholar] [CrossRef] [PubMed]
- Mechoulam, R.; Hanus, L. Cannabidiol: An overview of some chemical and pharmacological aspects. Part I: Chemical aspects. Chem. Phys. Lipids 2002, 121, 35–43. [Google Scholar] [CrossRef] [PubMed]
- Bergamaschi, M.M.; Queiroz, R.H.; Zuardi, A.W.; Crippa, J.A. Safety and side effects of cannabidiol, a Cannabis sativa constituent. Curr. Drug Saf. 2011, 6, 237–249. [Google Scholar] [CrossRef]
- Navarro, G.; Varani, K.; Lillo, A.; Vincenzi, F.; Rivas-Santisteban, R.; Raïch, I.; Reyes-Resina, I.; Ferreiro-Vera, C.; Borea, P.A.; Sánchez de Medina, V.; et al. Pharmacological data of cannabidiol- and cannabigerol-type phytocannabinoids acting on cannabinoid CB(1), CB(2) and CB(1)/CB(2) heteromer receptors. Pharmacol. Res. 2020, 159, 104940. [Google Scholar] [CrossRef]
- Sionov, R.V.; Steinberg, D. Anti-microbial activity of phytocannabinoids and endocannabinoids in the light of their physiological and pathophysiological roles. Biomedicines 2022, 10, 631. [Google Scholar] [CrossRef]
- Atalay, S.; Jarocka-Karpowicz, I.; Skrzydlewska, E. Antioxidative and anti-inflammatory properties of Cannabidiol. Antioxidants 2019, 9, 21. [Google Scholar] [CrossRef]
- Zuardi, A.W.; Crippa, J.A.; Hallak, J.; Bhattacharyya, S.; Atakan, Z.; Martín-Santos, R.; McGuire, P.K.; Guimarães, F.S. A critical review of the antipsychotic effects of cannabidiol: 30 years of a translational investigation. Curr. Pharm. Des. 2012, 18, 5131–5140. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Katchan, V.; David, P.; Shoenfeld, Y. Cannabinoids and autoimmune diseases: A systematic review. Autoimmun. Rev. 2016, 15, 513–528. [Google Scholar] [CrossRef] [PubMed]
- Saleemi, M.A.; Yahaya, N.; Zain, N.N.M.; Raoov, M.; Yong, Y.K.; Noor, N.S.; Lim, V. Antimicrobial and cytotoxic effects of Cannabinoids: An updated review with future perspectives and current challenges. Pharmaceuticals 2022, 15, 1228. [Google Scholar] [CrossRef] [PubMed]
- Appendino, G.; Gibbons, S.; Giana, A.; Pagani, A.; Grassi, G.; Stavri, M.; Smith, E.; Rahman, M.M. Antibacterial cannabinoids from Cannabis sativa: A structure-activity study. J. Nat. Prod. 2008, 71, 1427–1430. [Google Scholar] [CrossRef] [PubMed]
- Blaskovich, M.A.T.; Kavanagh, A.M.; Elliott, A.G.; Zhang, B.; Ramu, S.; Amado, M.; Lowe, G.J.; Hinton, A.O.; Pham, D.M.T.; Zuegg, J.; et al. The antimicrobial potential of cannabidiol. Commun. Biol. 2021, 4, 7. [Google Scholar] [CrossRef] [PubMed]
- Stahl, V.; Vasudevan, K. Comparison of efficacy of Cannabinoids versus commercial oral care products in reducing bacterial content from dental plaque: A preliminary observation. Cureus 2020, 12, e6809. [Google Scholar] [CrossRef] [PubMed]
- Wassmann, C.S.; Højrup, P.; Klitgaard, J.K. Cannabidiol is an effective helper compound in combination with bacitracin to kill Gram-positive bacteria. Sci. Rep. 2020, 10, 4112. [Google Scholar] [CrossRef] [Green Version]
- Feldman, M.; Sionov, R.V.; Mechoulam, R.; Steinberg, D. Anti-biofilm activity of Cannabidiol against Candida albicans. Microorganisms 2021, 9, 441. [Google Scholar] [CrossRef]
- Vasudevan, K.; Stahl, V. Cannabinoids infused mouthwash products are as effective as chlorhexidine on inhibition of total-culturable bacterial content in dental plaque samples. J. Cannabis Res. 2020, 2, 20. [Google Scholar] [CrossRef]
- Pitts, N.B.; Zero, D.T.; Marsh, P.D.; Ekstrand, K.; Weintraub, J.A.; Ramos-Gomez, F.; Tagami, J.; Twetman, S.; Tsakos, G.; Ismail, A. Dental caries. Nat. Rev. Dis. Primers 2017, 3, 17030. [Google Scholar] [CrossRef]
- Featherstone, J.D. Dental caries: A dynamic disease process. Aust. Dent. J. 2008, 53, 286–291. [Google Scholar] [CrossRef] [PubMed]
- Ahirwar, S.S.; Gupta, M.; Snehi, S.K. Dental caries and Lactobacillus: Role and ecology in the oral cavity. Int. J. Pharm. Sci. Res. 2019, 11, 4818–4829. [Google Scholar]
- Gilbert, K.; Joseph, R.; Vo, A.; Patel, T.; Chaudhry, S.; Nguyen, U.; Trevor, A.; Robinson, E.; Campbell, M.; McLennan, J.; et al. Children with severe early childhood caries: Streptococci genetic strains within carious and white spot lesions. J. Oral Microbiol. 2014, 6. [Google Scholar] [CrossRef] [Green Version]
- Salman, H.A.; Senthilkumar, R.; Imran, K.; Selvam, K.P. Isolation and typing of Streptococcus mutans and Streptococcus sobrinus from caries-active subjects. Contemp. Clin. Dent. 2017, 8, 587–593. [Google Scholar] [CrossRef] [PubMed]
- Jakubovics, N.S.; Goodman, S.D.; Mashburn-Warren, L.; Stafford, G.P.; Cieplik, F. The dental plaque biofilm matrix. Periodontol. 2000 2021, 86, 32–56. [Google Scholar] [CrossRef] [PubMed]
- Mirghani, R.; Saba, T.; Khaliq, H.; Mitchell, J.; Do, L.; Chambi, L.; Diaz, K.; Kennedy, T.; Alkassab, K.; Huynh, T.; et al. Biofilms: Formation, drug resistance and alternatives to conventional approaches. AIMS Microbiol. 2022, 8, 239–277. [Google Scholar] [CrossRef] [PubMed]
- Sionov, R.V.; Steinberg, D. Targeting the holy triangle of quorum sensing, biofilm formation, and antibiotic resistance in pathogenic bacteria. Microorganisms 2022, 10, 1239. [Google Scholar] [CrossRef]
- Lasserre, J.F.; Brecx, M.C.; Toma, S. Oral microbes, biofilms and their role in periodontal and peri-implant diseases. Materials 2018, 11, 1802. [Google Scholar] [CrossRef] [Green Version]
- Li, X.; Kolltveit, K.M.; Tronstad, L.; Olsen, I. Systemic diseases caused by oral infection. Clin. Microbiol. Rev. 2000, 13, 547–558. [Google Scholar] [CrossRef]
- Damle, S.G. Competence and transformation of oral Streptococcus sobrinus in dental caries. Contemp. Clin. Dent. 2018, 9, S195–S196. [Google Scholar] [CrossRef]
- Aqawi, M.; Sionov, R.V.; Gallily, R.; Friedman, M.; Steinberg, D. Anti-bacterial properties of Cannabigerol toward Streptococcus mutans. Front. Microbiol. 2021, 12, 656471. [Google Scholar] [CrossRef] [PubMed]
- Aqawi, M.; Sionov, R.V.; Gallily, R.; Friedman, M.; Steinberg, D. Anti-biofilm activity of Cannabigerol against Streptococcus mutans. Microorganisms 2021, 9, 2031. [Google Scholar] [CrossRef] [PubMed]
- Aqawi, M.; Steinberg, D.; Feuerstein, O.; Friedman, M.; Gingichashvili, S. Cannabigerol effect on Streptococcus mutans biofilms-A computational approach to confocal image analysis. Front. Microbiol. 2022, 13, 880993. [Google Scholar] [CrossRef] [PubMed]
- Guo, L.; Hu, W.; He, X.; Lux, R.; McLean, J.; Shi, W. Investigating acid production by Streptococcus mutans with a surface-displayed pH-sensitive green fluorescent protein. PLoS ONE 2013, 8, e57182. [Google Scholar] [CrossRef] [Green Version]
- Li, J.; Wu, T.; Peng, W.; Zhu, Y. Effects of resveratrol on cariogenic virulence properties of Streptococcus mutans. BMC Microbiol. 2020, 20, 99. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Dawes, C. What is the critical pH and why does a tooth dissolve in acid? J. Can. Dent. Assoc. 2003, 69, 722–724. [Google Scholar]
- Krzyściak, W.; Jurczak, A.; Kościelniak, D.; Bystrowska, B.; Skalniak, A. The virulence of Streptococcus mutans and the ability to form biofilms. Eur. J. Clin. Microbiol. Infect. Dis. 2014, 33, 499–515. [Google Scholar] [CrossRef] [Green Version]
- Høiby, N.; Bjarnsholt, T.; Givskov, M.; Molin, S.; Ciofu, O. Antibiotic resistance of bacterial biofilms. Int. J. Antimicrob. Agents 2010, 35, 322–332. [Google Scholar] [CrossRef] [Green Version]
- Hall-Stoodley, L.; Stoodley, P. Biofilm formation and dispersal and the transmission of human pathogens. Trends Microbiol. 2005, 13, 7–10. [Google Scholar] [CrossRef]
- Gomez, G.F.; Huang, R.; Eckert, G.; Gregory, R.L. Effect of phototherapy on the metabolism of Streptococcus mutans biofilm based on a colorimetric tetrazolium assay. J. Oral Sci. 2018, 60, 242–246. [Google Scholar] [CrossRef] [Green Version]
- Strahl, H.; Hamoen, L.W. Membrane potential is important for bacterial cell division. Proc. Natl. Acad. Sci. USA 2010, 107, 12281–12286. [Google Scholar] [CrossRef] [Green Version]
- Benarroch, J.M.; Asally, M. The Microbiologist’s guide to membrane potential dynamics. Trends Microbiol. 2020, 28, 304–314. [Google Scholar] [CrossRef] [PubMed]
- Vanhauteghem, D.; Janssens, G.P.; Lauwaerts, A.; Sys, S.; Boyen, F.; Cox, E.; Meyer, E. Exposure to the proton scavenger glycine under alkaline conditions induces Escherichia coli viability loss. PLoS ONE 2013, 8, e60328. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Spindler, E.C.; Hale, J.D.; Giddings, T.H., Jr.; Hancock, R.E.; Gill, R.T. Deciphering the mode of action of the synthetic antimicrobial peptide Bac8c. Antimicrob. Agents Chemother. 2011, 55, 1706–1716. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Schneider-Rayman, M.; Steinberg, D.; Sionov, R.V.; Friedman, M.; Shalish, M. Effect of epigallocatechin gallate on dental biofilm of Streptococcus mutans: An in vitro study. BMC Oral Health 2021, 21, 447. [Google Scholar] [CrossRef]
- Schwalbe, R.; Steele-Moore, L.; Goodwin, A.C. Antimicrobial Susceptibility Testing Protocols; CRC Press: Boca Raton, FL, USA, 2007. [Google Scholar]
- Sionov, R.V.; Tsavdaridou, D.; Aqawi, M.; Zaks, B.; Steinberg, D.; Shalish, M. Tooth mousse containing casein phosphopeptide-amorphous calcium phosphate prevents biofilm formation of Streptococcus mutans. BMC Oral Health 2021, 21, 136. [Google Scholar] [CrossRef]
- Feldman, M.; Ginsburg, I.; Al-Quntar, A.; Steinberg, D. Thiazolidinedione-8 alters symbiotic relationship in C. albicans-S. mutans dual species biofilm. Front. Microbiol. 2016, 7, 140. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Banerjee, S.; Sionov, R.V.; Feldman, M.; Smoum, R.; Mechoulam, R.; Steinberg, D. Anandamide alters the membrane properties, halts the cell division and prevents drug efflux in multidrug resistant Staphylococcus aureus. Sci. Rep. 2021, 11, 8690. [Google Scholar] [CrossRef]
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Barak, T.; Sharon, E.; Steinberg, D.; Feldman, M.; Sionov, R.V.; Shalish, M. Anti-Bacterial Effect of Cannabidiol against the Cariogenic Streptococcus mutans Bacterium: An In Vitro Study. Int. J. Mol. Sci. 2022, 23, 15878. https://doi.org/10.3390/ijms232415878
Barak T, Sharon E, Steinberg D, Feldman M, Sionov RV, Shalish M. Anti-Bacterial Effect of Cannabidiol against the Cariogenic Streptococcus mutans Bacterium: An In Vitro Study. International Journal of Molecular Sciences. 2022; 23(24):15878. https://doi.org/10.3390/ijms232415878
Chicago/Turabian StyleBarak, Tamar, Eden Sharon, Doron Steinberg, Mark Feldman, Ronit Vogt Sionov, and Miriam Shalish. 2022. "Anti-Bacterial Effect of Cannabidiol against the Cariogenic Streptococcus mutans Bacterium: An In Vitro Study" International Journal of Molecular Sciences 23, no. 24: 15878. https://doi.org/10.3390/ijms232415878