Periodontitis is associated with shifts in the balance of the subgingival microbiome. Many species that predominate in disease have not been isolated from healthy sites, raising questions as to the origin of these putative pathogens. The study aim was to determine whether periodontal pathogens could be enriched from pooled saliva, plaque and tongue samples from dentally-healthy adult volunteers using growth media that simulate nutritional aspects of the inflamed subgingival environment. The microbiome was characterised before and after enrichment using established metagenomic approaches, and the data analysed bioinformatically to identify major functional changes. After three weeks, there was a shift from an inoculum in whichStreptococcus,Haemophilus,Neisseria,VeillonellaandPrevotellaspecies predominated to biofilms comprising an increased abundance of taxa implicated in periodontitis, includingPorphyromonas gingivalis,Fretibacterium fastidiosum,Filifactor alocis,Tannerella forsythia, and severalPeptostreptococcusandTreponemaspp., with concomitant decreases in health-associated species. Sixty-four species were present after enrichment that were undetectable in the inoculum, includingJonquetella anthropi,Desulfovibrio desulfuricansandDialister invisus. These studies support the Ecological Plaque Hypothesis, providing evidence that putative periodontopathogens are present in health at low levels, but changes to the subgingival nutritional environment increase their competitiveness and drive deleterious changes to biofilm composition.

Veillonella spp. are predominant bacteria found in all oral biofilms. In this study, a metatranscriptomic approach was used to investigate the gene expression levels of three oral Veillonella spp. (V. parvula, V. dispar and V. atypica) in whole stimulated saliva from caries-free volunteers and in carious lesions (n=11 for each group). In the lesions the greatest proportion of reads were assigned to V. parvula and genes with the highest level of expression in carious samples were those coding for membrane transport systems. All three Veillonella spp. increased expression of genes involved in the catabolism of lactate and succinate, notably the alpha- and beta-subunits of L(+)-tartrate dehydratase (EC 4.2.1.32). There was also significantly increased expression of histidine biosynthesis pathway in V. parvula, suggesting higher intra-cellular levels of histidine that could provide intra-cellular buffering capacity and, therefore, assist survival in the acidic environment. Various other systems such as potassium uptake systems were also up regulated that may aid in the survival and proliferation of V. parvula in carious lesions.

Actinomyces spp. gene expression in root caries lesions

Following gingival recession, which increases with age, the root surface becomes exposed, creating new environments for microbial colonization and biofilm formation. The formation of root surface biofilms is influenced by the availability and composition of saliva and gingival crevicular fluid; they provide components for the conditioning film (acquired root surface pellicle) and also act as a source of nutrients. The early bacterial colonizers of the root surface are similar to those found on the enamel, and Gram-positive species such as Streptococcus sanguinis, S. oralis, S. mitis, and Actinomyces species predominate. The root surface has a lower mineral and higher organic content than enamel, and so is more vulnerable to demineralization. The characterization of the microbiota associated with root surface lesions is still ongoing. Traditional culture-based studies have implicated species such as mutans streptococci, lactobacilli, bifidobacteria, and Actinomyces species, while molecular-based studies have provided evidence for a more complex microbiota with many Gram-negative and anaerobic bacteria being detected in addition to the more conventional cariogenic organisms. Ecological concepts have been applied to explain the microbial etiology of root caries. The acidic environment generated from the fermentation of dietary sugars selects saccharolytic bacteria that can preferentially grow and metabolize under low pH conditions, and then proteolytic Gram-negative species are selected when the dentin is exposed and collagen and other proteins become accessible to be catabolized. These species act in concert to degrade the inorganic and organic components of the dental tissues.

The present study aimed to compare the absorption and transport patterns of three main betalains, betanin, vulgaxanthin I and indicaxanthin, into intestinal epithelial cells and to assess their distinct molecular effects on inflammatory and redox-related cell signalling in association with their radial scavenging potencies. All three betalains showed anti-inflammatory effects (5–80 μM), reflected by attenuated transcription of pro-inflammatory mediators such as cyclooxygenase-2 and inducible NO-synthase. Concomitant increases in antioxidant enzymes such as heme oxygenase-1 were only observed for betanin. Moreover, betanin uniquely demonstrated a potent dose-dependent radical scavenging activity in EPR and cell-based assays. Results also indicated overall low permeability for the three betalains with P_app of 4.2–8.9 × 10⁻⁷ cm s⁻¹. Higher absorption intensities of vulgaxanthin and indicaxanthin may be attributed to smaller molecular sizes and greater lipophilicity. In conclusion, betanin, vulgaxanthin I and indicaxanthin have differentially contributed to lowering inflammatory markers and mitigating oxidative stress, implying the potential to ameliorate inflammatory intestinal disease. Compared with two betaxanthins, the greater efficacy of betanin in scavenging radical and promoting antioxidant response might, to some extent, compensate for its poorer absorption efficiency, as demonstrated by the Caco-2 cell model.