Abstract
The worrying rise in antibiotic resistances emphasizes the need to seek new approaches for treating and preventing periodontal diseases. The purpose of this study was to evaluate the antibacterial and anti-biofilm activity of cranberry in a validated in vitro biofilm model. After chemical characterization of a selected phenolic-rich cranberry extract, its values for minimum inhibitory concentration and minimum bactericidal concentration were calculated for the six bacteria forming the biofilm (Streptococcus oralis, Actinomyces naeslundii, Veillonella parvula, Fusobacterium nucleatum, Porphyromonas gingivalis, and Aggregatibacter actinomycetemcomitans). Antibacterial activity of the cranberry extract in the formed biofilm was evaluated by assessing the reduction in bacteria viability, using quantitative polymerase chain reaction (qPCR) combined with propidium monoazide (PMA), and by confocal laser scanning microscopy (CLSM), and anti-biofilm activity by studying the inhibition of the incorporation of different bacteria species in biofilms formed in the presence of the cranberry extract, using qPCR and CLSM. In planktonic state, bacteria viability was significantly reduced by cranberry (p < 0.05). When growing in biofilms, a significant effect was observed against initial and early colonizers (S. oralis (p ≤ 0.017), A. naeslundii (p = 0.006) and V. parvula (p = 0.010)) after 30 or 60 s of exposure, while no significant effects were detected against periodontal pathogens (F. nucleatum, P. gingivalis or A. actinomycetemcomitans (p > 0.05)). Conversely, cranberry significantly (p < 0.001 in all cases) interfered with the incorporation of five of the six bacteria species during the development of 6 h-biofilms, including P. gingivalis, A. actinomycetemcomitans, and F. nucleatum. It was concluded that cranberry had a moderate antibacterial effect against periodontal pathogens in biofilms, but relevant anti-biofilm properties, by affecting bacteria adhesion in the first 6 h of development of biofilms.
Highlights
Dental biofilm-organized periodontal pathogens are the primary etiological factor of periodontal diseases, which are one of the most prevalent conditions affecting human beings [1]
The present study has confirmed the antibacterial capacity of cranberry extracts against the six bacteria species (S. oralis CECT 907T, V. parvula NCTC 11810, A. naeslundii ATCC 19039, F. nucleatum DMSZ 20482, A. actinomycetemcomitans DSMZ 8324, and P. gingivalis ATCC 33277) tested in planktonic state; this findings contradict, at least partially, those of La and co-workers [50], who concluded that A-type proanthocyanidins did not present any effect on P. gingivalis in planktonic state
The results of the present study indicate that, when testing bacteria organized in biofilms, bacteria viability was affected by exposure to the cranberry extract at 20 mg mL−1 after 30 and 60 s of exposure
Summary
Dental biofilm-organized periodontal pathogens (including Porphyromonas gingivalis and Aggregatibacter actinomycetemcomitans) are the primary etiological factor of periodontal diseases, which are one of the most prevalent conditions affecting human beings [1]. Cranberry (Vaccinium macrocarpum) compounds, including phenolic acids, proanthocyanidins (A-type proanthocyanidins), anthocyanins, organic acids, and their microbial-derived metabolites [13], selectively inhibit the growth of intestinal pathogens such as Staphylococcus strains and Salmonella enterica [14], reduce Escherichia coli colonization of the urinary tract [15,16,17], restrict the virulence of Pseudomonas aeruginosa [18,19], present anti-oxidant potential [20], anti-adhesion of Gram-negative and Gram-positive bacteria [21,22], and anti-motility [23,24] They may be associated with relevant health benefits, including a decreased risk of cardiovascular disease-related mortality [25], prevention of type 2 diabetes mellitus [26], and potential anti-cancer properties [27,28]
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