Antimicrobial activity of extract from GRAS plant species agains oral pathogenic microorganisms

Microbial oral diseases such as caries and periodontitis are among the most frequent human infections. Conventional chemical antiseptics used for their treatment and prevention often produce adverse side-effects, which restrict their long-term use. Although plants are considered as perspective sources of novel antimicrobial compounds, little is still known about their inhibitory properties against oral pathogens and about their safety while used on a daily basis. The objective of this study was to investigate in vitro growth-inhibitory effects of ethanol Generally Recognised as Safe (GRAS) plant and supercritical CO2 hop extracts on planktonic cultures of cariogenic, periodontal and candidal human pathogens, namely Aggregatibacter actinomycetemcomitans, Bifidobacterium dentium, B. longum, Candida albicans, Eikenella corrodens, Fusobacterium nucleatum subsp. vincentii, Lactobacillus salivarius subsp. salivarius, Porphyromonas gingivalis, Streptococcus mutans, S. salivarius subsp. salivarius and S. sobrinus using the broth microdilution method. The findings showed that ethanol extracts of all 109 GRAS plant species inhibited the growth of at least one microorganism under study with minimum inhibitory concentration (MIC) equal or lower than 4096 ug/mL. However, only six of them possessed very interesting antiseptic potential against the oral pathogens with MIC < 100 ug/mL. Additionally, three GRAS plant species showed good inhibitory activity with MIC = 128 ug/mL. As far as the particular results are concerned, the best antiseptic effect was observed for both Humulus lupulus CO2 supercritical and ethanol extracts that inhibited the growth of all microorganisms at MICs higher or equal to 8 ug/mL and MICs higher or equal to 16 ug/mL, respectively. Hence, the CO2 supercritical extraction proved to be superior for extraction of active constituents of H. lupulus. Further, the ethanol extracts of Capsicum annuum and C. frutescens showed significant antiseptic potential against S. sobrinus and S. salivarius (MICs from 64 to 128 ug/mL). The oral streptococci were also inhibited by Zanthoxylum clava-herculis (MIC higher or equal to 64 ug/mL), Helichrysum angustifolium and Myristica fragrans (MIC higher or equal to 128 ug/mL) which further showed the antimicrobial activity against F. nucleatum (MIC = 64 ug/mL). Interesting inhibitory effects exhibited an extract of Punica granatum against C. albicans (MIC = 128 ug/mL) and F. nucleatum (MIC = 64 ug/mL). Moreover, the growth of F. nucleatum was inhibited by extracts of Pimenta officinalis and Thea sinensis (MIC = 128 ug/mL). The chemical analysis of the CO2 supercritical H. lupulus extracts revealed that alpha and beta bitter acids represented the two major groups of constituents. Cohumulone was the predominant compound of the alpha acids, whereas colupulone was the prevalent constituent of the beta acids. Our results suggest that the six GRAS plant species, namely C. annuum, C. frutescens, H. lupulus, M. fragrans, P. granatum and Z. clava-herculis have good potential to become new safe antiseptic agents that might be used for incorporation into oral care products such as toothpastes and mouthrinses