Abstract
BackgroundExcessive biofilm formation on surfaces in the oral cavity is amongst the main reasons for severe infection development like periodontitis and peri-implantitis. Mechanical biofilm removal as well as the use of adjuvant antiseptics supports the prevention of pathogenic biofilm formation. Recently, the antibacterial effect of the oral care product REPHA-OS®, based on medicinal plant extracts and essential oils, has been demonstrated on oral pathogens grown on agar plates. In the present study, the effectiveness of the product on medical relevant oral biofilm development should be demonstrated for the first time.MethodsAn established in vitro oral multispecies biofilm, composed of Streptococcus oralis, Actinomyces naeslundii, Veillonella dispar and Porphyromonas gingivalis, was used to analyze the antibacterial effect of different REPHA-OS® concentrations on planktonic bacteria, biofilm formation and mature biofilms. It was quantified using metabolic activity assays and live/dead fluorescence staining combined with three-dimensional confocal laser-scanning microscopy. Additionally, effects on species distribution inside the biofilm were assessed by means of quantitative real-time PCR.ResultsREPHA-OS® showed statistically significant antimicrobial effects on all stages of biofilm development: a minimal inhibitory concentration of 5% could be detected for both, for planktonic bacteria and for biofilm formation. Interestingly, only a slightly higher concentration of 10% was necessary to completely kill all bacteria in mature biofilms also. In contrast, an influence on the biofilm matrix or the species distribution could not be observed. The effect could be attributed to the herbal ingredients, not to the contained ethanol.ConclusionThe strong antibacterial effect of REPHA-OS® on different stages of oral biofilm development strengthens its application as an alternative adjuvant in oral care therapies.
Highlights
Excessive biofilm formation on surfaces in the oral cavity is amongst the main reasons for severe infection development like periodontitis and peri-implantitis
Antibacterial effect of REPHA‐OS® on planktonic bacteria The effect of REPHA-OS® on planktonic bacteria after 24-h incubation was investigated by measuring the optical density to determine bacterial growth (Fig. 1a) and BacTiter-GloTM assay to quantify the amount of adenosine triphosphate (ATP) as parameter for metabolic activity (Fig. 1b)
We have investigated the effect of REPHA-OS® on colony growth on agar plates of single periodontal pathogens and halitosis-associated bacteria (Aggregatibacter actinomycetemcomitans, Fusobacterium nucleatum, Prevotella intermedia, P. gingivalis and Solobacterium moorei) [21]
Summary
Excessive biofilm formation on surfaces in the oral cavity is amongst the main reasons for severe infection development like periodontitis and peri-implantitis. If the bacterial biomass excessively increases, e.g., due to reduced oral hygiene or systemic precondition, the bacterial species distribution shifts and the amount of pathogens increases [7] These dysbiotic biofilms interfere with the host immune system and are often the reason for severe local infections like periodontitis and peri-implantitis [8]. Treatment of these oral infections is difficult due to the specific properties of biofilms: the biofilm matrix serves as diffusion barrier for antibacterial substances and the altered gene expression pattern effectively protects against the immune response of the host [9]. Prevention of an uncontrolled growth of bacteria is essential for prevention of infections in the oral cavity
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