Caries Experience in Elderly People Consuming a Milk-Based Drink Nutritional Supplement: A Cross-Sectional Study.

Nutritional supplements have been recommended to cope with malnutrition in elderly persons. In Chile, the Supplementary Nutrition Program for the Elderly (PACAM, for its Spanish acronym) consists in a monthly distribution of a low-fat-milk-based drink that contains 8% sucrose. The aim of this study was to determine whether older persons consuming the milk-based drink have a higher caries experience when compared to those not receiving the supplement. A cross-sectional study was conducted in the Maule Region in Chile. The representative sample comprised two groups: (a) PACAM consumers (CS) (n = 60) and (b) nonconsumers (NCS) (n = 60). Participants received intraoral examination and coronal (DMFT/DMFS) and root caries (RCI index) experience were recorded. Additionally, questionnaires regarding acceptability and consumption habits of PACAM and a 24-h diet recall were applied. The influence of predictors was calculated using binary logistic regression for a dichotomized DMFS and Poisson Regression for root caries lesions. A p value <0.05 was considered significant. No differences were detected in food consumption patterns between study groups (p > 0.05). CS participants had increased dairy product consumption. Higher DMFS mean value was observed in the CS (85.35 ± 39.0) compared with NCS (77.28 ± 28.9) (p = 0.043). The multivariate analysis showed nonconsumers of the milk-based product (ß = -0.41, p = 0.02) are less likely to have root surfaces affected by caries. Additionally, CS show higher RCI, compared to nonconsumers (ß = -0.17, p = 0.02). Daily consumption of a PACAM's milk-based drink supplement seems to increase coronal and root caries risk. Based on these results, composition modification of milk-based drinks with added sucrose appears mandatory.

Spatial mapping of polymicrobial communities reveals a precise biogeography associated with human dental caries.

Tooth decay (dental caries) is a widespread human disease caused by microbial biofilms. Streptococcus mutans, a biofilm-former, has been consistently associated with severe childhood caries; however, how this bacterium is spatially organized with other microorganisms in the oral cavity to promote disease remains unknown. Using intact biofilms formed on teeth of toddlers affected by caries, we discovered a unique 3D rotund-shaped architecture composed of multiple species precisely arranged in a corona-like structure with an inner core of S. mutans encompassed by outer layers of other bacteria. This architecture creates localized regions of acidic pH and acute enamel demineralization (caries) in a mixed-species biofilm model on human teeth, suggesting this highly ordered community as the causative agent. Notably, the construction of this architecture was found to be an active process initiated by production of an extracellular scaffold by S. mutans that assembles the corona cell arrangement, encapsulating the pathogen core. In addition, this spatial patterning creates a protective barrier against antimicrobials while increasing bacterial acid fitness associated with the disease-causing state. Our data reveal a precise biogeography in a polymicrobial community associated with human caries that can modulate the pathogen positioning and virulence potential in situ, indicating that micron-scale spatial structure of the microbiome may mediate the function and outcome of host-pathogen interactions.

Cariogenicity of a Milk-Based Drink Used as a Dietary Supplement for Older Adults Using a Root Caries Experimental Model.

The aim of this study was to evaluate the cariogenicity of a milk-based drink intended for older adults that was used as part of a governmental initiative in Chile to improve their nutritional conditions. This drink contains a high concentration of sugars, which can contribute to root caries development. To test this hypothesis, an experimental biofilm/caries model was used. Dentin slabs were used to grow biofilms of Streptococcus mutans UA159. Slabs/biofilms were exposed 3× per day to bovine milk with different fat content, the milk-based drink, and the milk-based drink supplemented with 10 g of sucrose added per serving. Slabs exposed to 10% sucrose or 0.9% NaCl were used as positive and negative controls, respectively. Biofilms were analyzed for bacterial counts and acidogenicity. Dentin demineralization was estimated by the loss of surface microhardness and integrated mineral loss. Results were compared by analysis of variance and Tukey's test. The milk-based drink showed higher acidogenicity than milk with its entire (whole) or reduced total fat content (skim). The milk-based drink supplemented with -sucrose had similar acidogenicity as the 10% sucrose positive control (p = 0.506). Whole milk exposure elicited lower bacterial counts than the positive control, the milk-based drink, and the milk-based drink supplemented with sucrose (p = 0.002; 0.006 and 0.014 respectively). Although skim milk induced higher demineralization than whole milk, both milk types produced lower demineralization than the milk-based drink. Regarding integrated mineral loss, demineralization induced by the milk-based drink and the milk-based drink supplemented with sucrose was similar to that induced by the positive control and skim milk (p > 0.05). Sugar-containing milk-based drinks used as dietary supplements for older adults may be highly cariogenic and could represent a potential risk for root caries.

In situ Effect of Arginine-Containing Dentifrice on Plaque Composition and on Enamel Demineralization under Distinct Cariogenic Conditions.

There is limited evidence that arginine-containing fluoridated dentifrices (AFD) have a better anticaries effect than regular fluoridated dentifrices (FD), especially in subjects at a higher risk for caries development. This study aimed to assess the effect of AFD on enamel demineralization and on the microbial and biochemical compositions of biofilm formed under different frequencies of sucrose exposure. It consisted of an in situ split-mouth design, where 12 adult volunteers who used FD for at least 2 months prior to the beginning of this study wore acrylic palatal appliances containing 4 bovine enamel specimens (1 pair at each side of the appliance) during 2 phases of 14 days each. FD slurry (3×/day) and 20% sucrose solution (4× and 8×/day) were dripped on the specimens during the first experimental phase. The same volunteers then used AFD during a 2-month washout period, followed by a second experimental phase where the AFD slurry and sucrose solution were applied onto a new subset of specimens. The percentage of enamel surface hardness loss (%SHL), the lesion depth (LD), the integrated mineral loss (IML), microbial counts on biofilms, the biomass, and inorganic and insoluble extracellular polysaccharide (IEPS) biofilm concentrations were determined. Higher %SHL, biomass, and IEPS and lower fluoride values were found at sucrose 8×/day exposure. Lower IEPS were found in the presence of AFD compared to FD. Similar %SHL, LD, and IML values were found between FD and AFD, irrespectively of the cariogenic challenge. The results suggest that AFD have an anticaries effect similar to that of regular FD.

Cariogenic Potential of Sucrose Associated with Maltodextrin on Dental Enamel.

Maltodextrin is a hydrolysate of cornstarch and has been widely used in the food industry associated with sucrose. The addition of starch can increase the cariogenic potential of sucrose; however, there are sparse data regarding the cariogenicity of sucrose associated with maltodextrin. Therefore, the aim of this study was to test in situ if maltodextrin could increase the cariogenic potential of sucrose. This was an in situ, randomized, crossover, split-mouth, and double-blind study. Volunteers wore palatal appliances containing bovine enamel blocks for 2 periods of 14 days. They dripped the following solutions on the enamel blocks 8 times per day: deionized distilled water (DDW), maltodextrin (M), sucrose + maltodextrin (S+M), or sucrose (S). At the end of each experimental period, biofilms were collected and analyzed for microbiological (mutans streptococci, lactobacilli, and total microorganisms counts) and biochemical (calcium, inorganic phosphate, fluoride, and insoluble extracellular polysaccharides concentrations) compositions. The enamel demineralization was assessed by microhardness. Treatments S and S+M resulted in a lower inorganic composition and higher concentration of insoluble extracellular polysaccharides in the biofilms, and higher enamel mineral loss compared to DDW and M. It can be concluded that the cariogenic potential of sucrose is not changed when this carbohydrate is associated with maltodextrin (dextrose equivalent 13-17).

Influence of TLR-2 in the immune response in the infection induced by fungus Sporothrix schenckii.

Toll-like receptors (TLRs) play an important role in immunity, since they bind to pathogen surface antigens and initiate the immune response. However, little is known about the role of TLR-2 in the recognition of S. schenckii and in the subsequent immune response. Therefore, the aim of this study was to evaluate the involvement of TLR-2 in the immune response induced by S. schenckii. C57BL/6 mice (WT) and C57BL/6 TLR-2 knockout (TLR-2-/-) were used to evaluate, over a period of 10 weeks of sporotrichotic infection, the influence of TLR-2 over macrophages production of IL-1ß, IL-12 and TNF-α, their stimulation level by NO release and the production of IFN -γ, IL-6, IL-17 and TGF-ß by spleen cells. The results showed that the production of pro-inflammatory mediators and NO, TLR-2 interference is striking, since its absence completely inhibited it. IL-17 production was independent of TLR-2. The absence of Th1 response in TLR2-/- animals was concomitant with IL-17 production. Therefore, it can be suggested that TLR-2 absence interferes with the course of the infection induced by the fungus S. schenckii.

Antimicrobial effects of drug-containing electrospun matrices on osteomyelitis-associated pathogens.

PURPOSE: To synthesize polydioxanone (PDS)-based drug delivery systems (hereafter referred to as "matrices") containing vancomycin (VANC) and/or rifampicin (RIF) and investigate their effect on the inhibition of biofilm growth containing osteomyelitis (OM)-associated pathogens. MATERIALS AND METHODS: PDS matrices were prepared by electrostatic spinning, and the drugs were incorporated as follows: group (G)1, 5wt%VANC; G2, 10wt%VANC; G3, 5wt%RIF; G4, 10wt%RIF; G5, 5wt%VANC+RIF; and G6, 10wt%VANC+RIF. A control group of pure PDS was also electrospun (G7). Biofilms formed by Staphylococcus aureus and S. epidermidis were grown on the electrospun matrices for 24 hours. The counts of viable cells were assessed after biofilm formation. The fiber morphology and biofilms were imaged using a scanning electron microscope. RESULTS: G5 and G6 and pure PDS (G7) had the lowest and highest mean number of viable cell counts, respectively (P < .05). Small and isolated clusters of bacteria with no mature biofilm present were found on G6. CONCLUSIONS: The results of the present study have provided evidence for the potential use of PDS-based matrices as an effective drug delivery system that could inhibit biofilm formation from OM-associated pathogens.

Biodegradable nanofibrous drug delivery systems: effects of metronidazole and ciprofloxacin on periodontopathogens and commensal oral bacteria.

OBJECTIVES: The purposes of this study were to fabricate biodegradable polydioxanone (PDS II®) electrospun periodontal drug delivery systems (hereafter referred to as matrices) containing either metronidazole (MET) or ciprofloxacin (CIP) and to investigate the effects of antibiotic incorporation on both periodontopathogens and commensal oral bacteria. MATERIALS AND METHODS: Fibrous matrices were processed from PDS polymer solution by electrospinning. Antibiotic-containing PDS solutions were prepared to obtain four distinct groups: 5 wt.% MET, 25 wt.% MET, 5 wt.% CIP, and 25 wt.% CIP. Pure PDS was used as a control. High-performance liquid chromatography (HPLC) was done to evaluate MET and CIP release. Dual-species biofilms formed by Lactobacillus casei (Lc) and Streptococcus salivarius (Ss) were grown on the surface of all electrospun matrices. After 4 days of biofilm growth, the viability of bacteria on biofilms was assessed. Additionally, antimicrobial properties were evaluated against periodontopathogens Fusobacterium nucleatum (Fn) and Aggregatibacter actinomycetemcomitans (Aa) using agar diffusion assay. RESULTS: A three-dimensional interconnected porous network was observed in the different fabricated matrices. Pure PDS showed the highest fiber diameter mean (1,158 ± 402 nm) followed in a descending order by groups 5 wt.% MET (1,108 ± 383 nm), 25 wt.% MET (944 ± 392 nm), 5 wt.% CIP (871 ± 309 nm), and 25 wt.% CIP (765 ± 288 nm). HPLC demonstrated that groups containing higher amounts (25 wt.%) of incorporated drugs released more over time, while those with lower levels (5 wt.%) the least. No inhibitory effect of the tested antibiotics was detected on biofilm formation by the tested commensal oral bacteria. Meanwhile, CIP-containing matrices inhibited growth of Fn and Aa. CONCLUSION: CIP-containing matrices led to a significant inhibition of periodontopathogens without negatively impairing the growth of periodontal beneficial bacteria. CLINICAL RELEVANCE: Based on the proven in vitro inhibition of periodontitis-related bacteria, future in vivo research using relevant animal models is needed to confirm the effectiveness of these drug delivery systems.

Role of TLR-2 and fungal surface antigens on innate immune response against Sporothrix schenckii.

Sporotrichosis is an infection caused by the dimorphic fungus Sporothrix schenckii. Toll-like receptors (TLRs) play an important role in immunity, since they bind to pathogen surface antigens and initiate the immune response. However, little is known about the role of TLR-2 and fungal surface antigens in the recognition of S. schenckii and in the subsequent immune response. This study aimed to evaluate the involvement of TLR-2 and fungal surface soluble (SolAg) and lipidic (LipAg) antigens in phagocytosis of S. schenckii and production of immune mediators by macrophages obtained from WT and TLR-2(-/-) animals. The results showed that TLR-2(-/-) animals had had statistical lower percentage of macrophages with internalized yeasts compared to WT. SolAg and LipAg impaired phagocytosis and immunological mediator production for both WT and TLR-2(-/-). The absence of TLR-2 led to lower production of the cytokines TNF-α, IL-1ß, IL-12 and IL-10 compared to WT animals. These results suggest a new insight in relation to how the immune system, through TLR-2, recognizes and induces the production of mediators in response to the fungus S. schenckii.

Farnesol delivery via polymeric nanoparticle carriers inhibits cariogenic cross-kingdom biofilms and prevents enamel demineralization.

Streptococcus mutans and Candida albicans are frequently detected together in the plaque from patients with early childhood caries (ECC) and synergistically interact to form a cariogenic cross-kingdom biofilm. However, this biofilm is difficult to control. Thus, to achieve maximal efficacy within the complex biofilm microenvironment, nanoparticle carriers have shown increased interest in treating oral biofilms in recent years. Here, we assessed the anti-biofilm efficacy of farnesol (Far), a hydrophobic antibacterial drug and repressor of Candida filamentous forms, against cross-kingdom biofilms employing drug delivery via polymeric nanoparticle carriers (NPCs). We also evaluated the effect of the strategy on teeth enamel demineralization. The farnesol-loaded NPCs (NPC+Far) resulted in a 2-log CFU/mL reduction of S. mutans and C. albicans (hydroxyapatite disc biofilm model). High-resolution confocal images further confirmed a significant reduction in exopolysaccharides, smaller microcolonies of S. mutans, and no hyphal form of C. albicans after treatment with NPC+Far on human tooth enamel (HT) slabs, altering the biofilm 3D structure. Furthermore, NPC+Far treatment was highly effective in preventing enamel demineralization on HT, reducing lesion depth (79% reduction) and mineral loss (85% reduction) versus vehicle PBS-treated HT, while NPC or Far alone had no differences with the PBS. The drug delivery via polymeric NPCs has the potential for targeting bacterial-fungal biofilms associated with a prevalent and costly pediatric oral disease, such as ECC.

Objective and quantitative assessment of caries lesion activity.

OBJECTIVES: Evaluate the ability of objectively measured specular reflection, roughness, and fluorescence change during dehydration to assess caries lesion activity. METHODS: One hundred ninety-five ground/polished 3 × 3 × 2 mm sound human enamel specimens were divided into three groups and demineralized using a multispecies microbial caries model for 3, 6, or 9 days; and then remineralized with 1100 ppm-F as NaF solution for 10 days using a pH-cyclic model. Reflection (amplitude: %), roughness (Ra: µm), fluorescence change during dehydration (ΔQ: %×mm2), and microfocus computed tomography [µ-CT: lesion volume (µm3)] were measured for sound, demineralized and remineralized enamel. The surface was hydrated and fluorescence images were acquired at 1 s intervals for 10 s (ΔQ10). During image acquisition, surface was dehydrated with continuous compressed air. Changes-in-ΔQ per second (ΔQD: %×mm2/sec) at 5 (ΔQD5) and 10 s (ΔQD10) were obtained. RESULTS: Reflection decreased from sound to demineralized groups (p < 0.0001); remineralized groups were higher than demineralized groups (p < 0.001), but not different from sound (p > 0.32). Roughness increased from sound to demineralized groups (p < 0.0001) and remineralized groups were also higher than sound (p < 0.0001). ΔQ10, ΔQD5 and ΔQD10 increased from sound to demineralized groups (p < 0.0001), and remineralized groups decreased compared to demineralized groups (p < 0.05), but was higher than sound (p < 0.0001). The correlations of µ-CT with reflection, roughness, and ΔQ10 were -0.63, 0.71, and 0.82, respectively (p < 0.0001). CONCLUSIONS: Reflection, roughness and ΔQ could distinguish between sound and demineralized enamel. Reflection and ΔQ were able to distinguish between demineralized and remineralized enamel. CLINICAL SIGNIFICANCE: Determination of caries activity, whether a lesion is active or inactive, is an essential and critical component of caries diagnosis. However, especially for enamel lesions, it is difficult to estimate without longitudinal follow-up. Reflection, roughness and fluorescence change during dehydration have the potential to measure caries lesion activity at a-single-appointment.

Salivary epithelial cells as model to study immune response against cutaneous pathogens.

The human skin not only provides passive protection as a physical barrier against external injury, but also mediates active surveillance via epidermal cell surface receptors that recognize and respond to potential invaders. Primary keratinocytes and immortalized cell lines, the commonly used sources to investigate immune responses of cutaneous epithelium are often difficult to obtain and/or potentially exhibit changes in cellular genetic make-up. Here we investigated the possibility of using salivary epithelial cells (SEC) to evaluate the host response to cutaneous microbes. Elevated secretion of IFN-γ and IL-12 was observed in the SEC stimulated with Staphylococcus aureus, a transient pathogen of the skin, as mono species biofilm as compared to SEC stimulated with a commensal microbe, the Staphylococcus epidermidis. Co-culture of the SEC with both microbes as dual species biofilm elicited maximum cytokine response. Stimulation with S. aureus alone but not with S. epidermidis alone induced maximum toll-like receptor-2 (TLR-2) expression in the SEC. Exposure to dual species biofilm induced a sustained upregulation of TLR-2 in the SEC for up to an hour. The data support novel application of the SEC as efficient biospecimen that may be used to investigate personalized response to cutaneous microflora.