Mechanistic Effects of E-Liquids on Biofilm Formation and Growth of Oral Commensal Streptococcal Communities: Effect of Flavoring Agents

Abstract Background: Vaping has become a global health concern. As research continues, more studies are beginning to question the relative safety of E-liquid flavoring additives. The oral cavity is the first site of exposure to E-liquid aerosol, making it critical for investigation. Because of the importance of commensal bacterial biofilms for oral health, we sought to explore the effects of E-liquids ± flavors on the formation and growth of single- and multi-species biofilms and to investigate the mechanism of inhibition. Methods: Quantitative and confocal biofilm analysis, death curves, and colony-forming units (CFU) were evaluated with flavorless and flavored (tobacco, menthol, cinnamon, strawberry, blueberry) E-liquids using four strains of oral commensal bacteria (Streptococcus gordonii, Streptococcus intermedius, Streptococcus mitis, and Streptococcus oralis). Results: All flavoring agents show a dose-dependent inhibition in the growth of single-species and multi-species biofilms. Furthermore, CFUs, death curves, and light microscopy show that flavoring agents have a bactericidal mode of inhibition on the growth of these oral streptococci. Conclusions: These results show that flavored, rather than unflavored, E-liquids are more detrimental to biofilm formation and growth of oral commensal bacteria. Consequently, E-liquid flavorings agents could pose risks to the oral microenvironment, and by extension, to systemic health.