Species-specific growth inhibitory effect of propolis

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Abstract

<p>The goal of dentistry is to prevent not only dental diseases but also systemic diseases by improving the oral environment. In this context, we focus on the improvement of oral microbiota. Various effects have been reported for propolis, a natural antimicrobial substance, which is used in health foods and folk medicines for periodontal diseases. We examined the effect of propolis on the oral microbiota using a next-generation sequencer. Gingival crevicular fluid was collected from periodontal pockets of more than 4 mm in three patients (two males and one female) at the Osaka Dental University Hospital, and the samples were cultured in SHI medium to obtain oral microbiota samples. Ethanol-extracted propolis (EEP; 0, 50 and 100 μg/mL; provided by Yamada Bee Farm, Okayama, Japan) was added to the samples and DNA was extracted after 24 h of incubation at 37°C with shaking under anaerobic conditions. Sequences of the V3-V4 region of the PCR-amplified 16S rRNA were decoded using the next-generation sequencer Miseq and changes in the microbiota after EEP treatment were analyzed. The effect of propolis on the growth of the periodontal bacteria Porphyromonas gingivalis (P. gingivalis) and Fusobacterium nucleatum (F. nucleatum) was then assessed by measuring the absorbance as optical density (OD) of liquid culture media and the colony forming units (CFUs) on solid media. EEP treatment did not show significant changes in the proportion and diversity of the cultured oral microbiota. However, EEP inhibited the growth of P. gingivalis in a dose-dependent manner, as evidenced in the experiments using OD measurements. In contrast, there was no significant effect on F. nucleatum proliferation. Furthermore, 100 μg/mL EEP reduced the number of P. gingivalis CFUs, but not that of F. nucleatum. These results suggest that EEP specifically inhibits the growth of the periodontal bacterium P. gingivalis without significantly altering the bacterial microbiota. (J Osaka Dent Univ 2021; 55: 83-89)</p>

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