Machine learning enabled multiplex detection of periodontal pathogens by surface-enhanced Raman spectroscopy.

Periodontitis is a chronic inflammation of the periodontium caused by a persistent bacterial infection, resulting in destruction of the supporting structures of teeth. Analysis of microbial composition in saliva can inform periodontal status. Actinobacillus actinomycetemcomitans (Aa), Porphyromonas gingivalis (Pg), and Streptococcus mutans (Sm) are among reported periodontal pathogens, and were used as model systems in this study. Our atomic force microscopic (AFM) study revealed that these pathogens are biological nanorods with dimensions of 0.6-1.1 µm in length and 500-700 nm in width. Current bacterial detection methods often involve complex preparation steps and require labeled reporting motifs. Employing surface-enhanced Raman spectroscopy (SERS), we revealed cell-type specific Raman signatures of these pathogens for label-free detection. It overcame the complexity associated with spectral overlaps among different bacterial species, relying on high signal-to-noise ratio (SNR) spectra carefully collected from pure species samples. To enable simple, rapid, and multiplexed detection, we harnessed advanced machine learning techniques to establish predictive models based on a large set of raw spectra of each bacterial species and their mixtures. Using these models, given a raw spectrum collected from a bacterial suspension, simultaneous identification of all three species in the test sample was achieved at 95.6 % accuracy. This sensing modality can be applied to multiplex detection of a broader range and a larger set of periodontal pathogens, paving the way for hassle-free detection of oral bacteria in saliva with little to no sample preparation.

Effect of nano-ceramic coating on surface property and microbial adhesion to poly(methyl methacrylate).

To improve surface properties of poly(methyl methacrylate) (PMMA) using nano-ceramic coatings and assess microbial adherence after long-term use of a chemical cleanser. Thirty-six PMMA samples were fabricated, polished and coated with a nano-thin TiO2 or mixed TiO2 /ZrO2 , with uncoated samples as controls. Six samples in each group (n = 12) were soaked in Polident denture cleaner 180 times for 30 min, while six were soaked in deionized water. Surface roughness of PMMA before and after being soaked in Polident was assessed. All samples were subsequently exposed to Candida albicans for 6 h and the adherent cells were determined by viable colony count. Two-way analysis of variance was performed for statistical analysis. No significant difference in surface roughness was noted between the uncoated and coated PMMA before soaking. After soaking, surface roughness of the uncoated PMMA increased from 0.164 to 0.532 µm (p < .05). No significant change was observed for TiO2 -coated (0.105-0.143 µm) or TiO2 /ZrO2 -coated PMMA (0.104-0.141 µm). Attachment of C. albicans to PMMA soaked in water showed significantly less attachment to both TiO2 -coated (1.4 × 103 cfu/ml) and TiO2 /ZrO2 -coated PMMA (1.6 × 103 cfu/ml) than to the uncoated PMMA (2.6 × 103 cfu/ml). After soaking in Polident, the uncoated PMMA had significantly less C. albicans attachment than coated samples. Less attachment was noted on the TiO2 /ZrO2 -coated PMMA then the TiO2 -coated samples (p < .05). Nano-ceramic TiO22 /ZrO2 coating of PMMA denture base material alters surface properties thus reduces oral microbial adhesion. It represents a promising alternative to the chemical disinfection for PMMA denture materials.

Downregulation of miRNA-26 in chronic periodontitis interferes with innate immune responses and cell migration by targeting phospholipase C beta 1.

AIM: To evaluate the potential role of miR-26 family members in periodontal pathogenesis by assessing innate immune responses to periopathic bacteria and regulation of cytoskeletal organization. MATERIALS AND METHODS: Expression of miR-26a-5p and miR-26b-5p was quantified in gingival biopsies derived from healthy and periodontally diseased subjects before and after non-surgical (scaling and root planing) therapy by RT-qPCR. Global pathway analysis and luciferase assays were performed for target identification and validation. Cytokine expression was assessed in miR-26a-5p transfected human oral keratinocytes upon stimulation with either live Porphyromonas gingivalis (Pg), Aggregatibacter actinomycetemcomitans or Pg lipopolysaccharide (LPS). Wound closure assays were performed in cells transfected with miR-26a-5p, while the impact on cytoskeletal organization was assessed by F-actin staining. RESULTS: miR-26a-5p and miR-26b-5p were downregulated in diseased gingiva and restored 4-6 weeks post-therapy to levels comparable with healthy subjects. Target validation assays identified phospholipase C beta 1 as a bona fide novel target exhibiting antagonistic expression pattern in disease and post-therapy cohorts. miR-26a-5p transfected cells secreted higher levels of cytokine/chemokines upon stimulation with periopathogens and demonstrated impaired cell migration and cytoskeletal rearrangement. CONCLUSIONS: Downregulated miR-26a-5p levels in periodontal inflammation may interfere with key cellular functions that may have significant implications for host defence and wound healing.

Comparison of collagen features of distinct types of caries-affected dentin.

OBJECTIVES: To compare the biodegradability, mechanical behavior, and physicochemical features of the collagen-rich extracellular matrix (ECM) of artificial caries-affected dentin (ACAD), natural caries-affected dentin (NCAD) and sound dentin (SD). METHODS: Dentin specimens from human molars were prepared and assigned into groups according to the type of dentin: ACAD, NCAD, or SD. ACAD was produced by incubation of demineralized SD with Streptococcus mutans in a chemically defined medium (CDM) with 1% sucrose for 7 days at 37 °C under anaerobic conditions. Specimens were assessed to determine collagen birefringence, biodegradability, mechanical behavior, and chemical composition. Data were individually processed and analyzed by ANOVA and post-hoc tests (α = 0.05). RESULTS: CDM-based biofilm challenge reduced loss, storage, and complex moduli in ACAD (p < 0.001), while the damping capacity remained unaffected (p = 0.066). Higher red and lower green birefringence were found in ACAD and NCAD when compared with SD (p < 0.001). Differently to ACAD, SD and NCAD presented higher biodegradability to exogenous proteases (p = 0.02). Chemical analysis of the integrated areas of characteristic bands that assess mineral quality (carbonate/phosphate and crystallinity index), mineral to matrix (phosphate/amide I) and post-translational modifications (amide III/CH2, pentosidine/CH2, and pentosidine/amide III) (p<0.05) showed that NCAD was significantly different from SD while ACAD exhibited intermediate values. CONCLUSIONS: CDM-based biofilm challenge produced a dentin ECM with decreased mechanical properties and increased collagen maturity. The compositional and structural conformation of the ACAD suggested that CDM-based biofilm challenge showed potential to produce artificial lesions by revealing a transitional condition towards mimicking critical features of NCAD. CLINICAL SIGNIFICANCE: This study highlights the importance of developing a tissue that mimics the features of natural caries-affected dentin ECM for in vitro studies. Our findings suggested the potential of a modified biofilm challenge protocol to produce and simulate a relevant substrate, such as caries-affected dentin.

In Vitro Biofilm Formation on Aryl Ketone Polymer (AKP), A New Denture Material, Compared with That on Three Traditional Dental Denture Materials.

Control of denture plaque biofilms is a practical approach to preventing persistent oral infections such as denture stomatitis. Objectives. This study compared in vitro biofilm attachment and growth on a new denture material, Ultaire® AKP, with that on traditional denture materials including cobalt chrome (CoCr), polymethyl methacrylate (PMMA), and polyoxymethylene (POM). Methods. Microbial biofilms were grown with cultures of Candida albicans, Streptococcus mutans UA159, or a mixed Streptococcus spp. (S. mutans 700610/Streptococcus sanguinis BAA-1455) for 6 hours in a static protocol or 24 hours in a dynamic protocol for each material. Adherent biofilm cells were removed, and viable colony-forming units (CFUs) were enumerated. Confocal microscopy of the 24-hour Streptococcus spp. biofilms was used to determine biofilm mass and roughness coefficients. Results. The rank order of C. albicans attachment after 6 hours was CoCr > PMMA ∗ > Ultaire® AKP ∗ ( ∗ vs CoCr, p ≤ 0.05), and that for 24-hour biofilm growth was CoCr > Ultaire® AKP ∗ > PMMA ∗ ( ∗ vs CoCr, p ≤ 0.05). The rank order of S. mutans biofilm attachment was CoCr > POM > Ultaire® AKP ∗ > PMMA ∗ ( ∗ vs CoCr, p ≤ 0.05), and that for the 24-hour Streptococcus spp. biofilm growth was POM > Ultaire® AKP > PMMA > CoCr ∗ ( ∗ vs POM, p ≤ 0.05). Confocal images revealed structural differences in Streptococcus spp. biofilms on CoCr compared with the other test materials. Significantly lower roughness coefficients of Streptococcus spp. biofilms on Ultaire® AKP were noted, suggesting that these biofilms were less differentiated. Ultaire® AKP promoted significantly less C. albicans and S. mutans biofilm attachment than CoCr at 6 hours and C. albicans growth at 24 hours. Streptococcus spp. biofilms on Ultaire® AKP were less differentiated than those on other test materials. Conclusion. In addition to its material strength, Ultaire® AKP represents an attractive option for denture material in removable partial dentures.

Gelatinolytic Activity and Adhesion Studies of Artificial Caries-affected Dentin do not Simulate Natural Caries.

PURPOSE: To investigate whether interfacial enzymatic activity and adhesion receptiveness of artificial caries-affected dentin (ACAD) simulate those of natural caries-affected dentin (NCAD). MATERIALS AND METHODS: Thirty dentin specimens were prepared from human molars to determine interfacial gelatinolytic activity using in situ zymography and adhesion experiments (micropermeability and bond strength [µTBS]). Groups were formed according to the type of dentin: artificial caries-affected dentin (ACAD), natural caries-affected dentin (NCAD), or sound dentin. ACAD was produced by incubating dentin with Streptococcus mutans in a chemically defined medium (CDM) with 1% sucrose for 7 days at 37°C under anaerobic conditions. CDM was replaced daily, and the sterility as well as the pH of the culture was monitored. Adhesion experiments employed Single Bond Universal (3M Oral Care) in self-etch mode. Data were individually processed and analyzed using ANOVA and post-hoc tests (α = 0.05). RESULTS: The enzymatic activity of ACAD was similar to that of sound dentin, but was lower than that of NCAD, which elicited the highest activity (p < 0.05). Interfacial micropermeability intensity at the hybrid layer or in underlying dentin (5 µm below the interface) was similar in all types of dentin (p > 0.05). On the other hand, substrate permeability was higher for NCAD than for ACAD. The highest sealing ability was detected in sound dentin. Bond strengths to ACAD were higher than to NCAD. However, the highest µTBS was observed in sound dentin (p < 0.05). CONCLUSION: Artificial caries-affected dentin simulated neither the gelatinolytic activity nor bonding receptiveness of natural caries-affected dentin.

Effect of Ultraviolet Radiation on Candida albicans Biofilm on Poly(methylmethacrylate) Resin.

PURPOSE: To investigate the effect of 254-nm ultraviolet light on the viability of Candida albicans biofilm on poly(methylmethacrylate). METHODS: Poly(methylmethacrylate) specimens (1 cm × 1 cm × 1 mm) were placed in 6-well culture plates. Each well contained 8 ml of 104 colony forming units/milliliter of C. albicans ATCC90028 and Sabouraud dextrose broth. Plates were incubated at 37°C for 24 hours. Specimens were then divided into 11 groups (n = 4): no treatment control groups, 3.8% sodium perborate immersion for 5 minutes (PP5m) and for 12 hours (PO12h), and 6 groups exposed to ultraviolet light for 5, 15, 30, 60, 120 or 300 seconds separately using UVP XX-15S series lamps. After sonication, cell suspensions were plated, and colony-forming units were counted. The relationship between survival of C. albicans and ultraviolet light irradiation energy exposure was analyzed and compared to the survival of sodium perborate groups. The effect of disinfection treatments and ultraviolet light energy exposure on C. albicans survival was analyzed with ANOVA (alpha = 0.05). RESULTS: There was a significant decrease in C. albicans survival with increasing ultraviolet light energy exposure (p = 0.00001, p < 0.05) Survival vs. immersion exposure time analysis of chemical disinfection showed no survival of C. albicans in groups PP5m and PO12h. C. albicans in the UV 300s group with energy of 210 mJ/cm2 (71 CFU/ml) showed a statistical difference from that of two chemical immersion groups (PP5m, PO12h = 0 CFU/ml) (p = 0.00001, p < 0.05). CONCLUSION: Ultraviolet 254 nm irradiation demonstrated a significant inhibition of C. albicans survival on poly(methylmethacrylate) samples. Ultraviolet light exposure of 300 seconds inhibited the survival of C. albicans close to the level of 3.8% sodium perborate treatment.

Effect of mechanically stimulated saliva on initial human dental biofilm formation.

This study evaluated the impact of mechanically stimulated saliva on initial bacterial colonization. Interaction between oral bacteria and both unstimulated and stimulated saliva was examined in vitro by laying labeled bacteria over SDS-PAGE-separated salivary proteins. The effects of chewing on in vivo biofilm, microbial composition, and spatial arrangement were examined in two human volunteers using an intraoral stent containing retrievable enamel chips. In vitro experiments showed that bacterial binding to proteins from stimulated saliva was lower than that to proteins from unstimulated saliva. Lack of binding activity was noted with Streptococcus mutans and Lactobacillus casei. Human Oral Microbe Identification Microarray (HOMIM) analyses revealed a consistent chewing-related increase in the binding of Streptococcus anginosus and Streptococcus gordonii. Immunofluorescence microscopy demonstrated the presence of multi-species colonies and cells bearing different serotypes of the coaggregation-mediating streptococcal cell-surface receptor polysaccharides (RPS). Differences in bacterial colonization were noted between the two volunteers, while the type 4 RPS-reactive serotype was absent in one volunteer. Cells reacting with antibody against Rothia or Haemophilus were prominent in the early biofilm. While analysis of the data obtained demonstrated inter-individual variations in both in vitro and in vivo bacterial binding patterns, stimulating saliva with multiple orosensory stimuli may modulate oral bacterial colonization of tooth surfaces.

Improving Polymethyl Methacrylate Resin Using a Novel Titanium Dioxide Coating.

PURPOSE: The objective of this study was to improve the surface characteristics of poly (methyl methacrylate) (PMMA) by developing a novel, thin film coating process and to characterize the resulting coated surface. MATERIALS AND METHODS: An atomic layer deposition (ALD) technique was developed to deposit a titanium dioxide (TiO2 ) nano-thin film on PMMA. The surface wettability for both coated and uncoated PMMA was determined by measuring water contact angle. Wear resistance was assessed using a mechanical tooth-brushing device with a 50 g load for 6000 strokes after 5 months of water storage. A denture cleanser challenge test was performed by using sonication in 3.8% sodium perborate for 1 hour with aged specimens. X-ray photoelectron spectroscopy (XPS) was used before and after the brushing test and challenge test to analyze the PMMA surface chemical composition. The mechanical strength of coated and uncoated PMMA was measured using a three-point bending test. Surface microbial interactions were also evaluated by assessing Candida albicans biofilm attachment. RESULTS: Nano-TiO2 coating (30 nm thick) was successfully deposited on PMMA at 65°C. After coating, water contact angle decreased from 70° to less than 5°. After brushing test, the coating remained intact. XPS analysis revealed no loss of TiO2 from coated specimens following brushing and denture cleanser sonication for 1 hour. There was no statistically significant difference in mechanical strength (MPa) (mean ± SD) between PMMA (139.4 ± 11.3) and TiO2 -PMMA (160.7 ± 37.1) (p = 0.0995). C. albicans attachment decreased by 63% to 77% on the coated PMMA surface. CONCLUSIONS: ALD is a promising technique to modify surface properties of PMMA and resulted in a stable adherent thin film. By depositing a TiO2 coating, PMMA surface properties may lead to significantly reduced microorganism adhesion and easier pathogen removal from PMMA. For patients who wear dentures, reducing the oral microbial biofilm burden using a TiO2 -coated PMMA surface could positively impact their oral and systemic health.

Growth Media Affect Assessment of Antimicrobial Activity of Plant-Derived Polyphenols.

This study aimed to investigate the effects of different microbial growth media on the laboratory assessment of antimicrobial activity of natural polyphenolic compounds. The inhibition of the tea polyphenol EGCG on growth of selected oral microorganisms was evaluated in complex media and a protein-free chemically defined medium (CDM). Other antimicrobial agents (polyphenolic grape seed extract, plant alkaloid berberine, methyl salicylate, and chlorhexidine gluconate) were also tested in the study. The presence of proteins and their effects on the antimicrobial activity of EGCG were investigated by the addition of BSA to the CDM. The MICs of EGCG against test oral microorganisms were 4 to 64 times higher in complex media than in CDM. The polyphenolic grape seed extract exhibited similar discrepancies. However, the MICs of the nonpolyphenolic compounds (berberine, methyl salicylate, and chlorhexidine) were not significantly different between the two growth media. The MIC of EGCG against S. mutans UA159 in CDM with added BSA was 16 times higher than that in CDM alone. Therefore, nonproteinaceous CDM should be used to avoid interference of proteins with the active ingredients when testing the antimicrobial activity of plant-derived polyphenolic compounds against microorganisms. This will also minimize the discrepancies noted in results obtained by different investigators.

Synergistic effects of tea catechin epigallocatechin gallate and antimycotics against oral Candida species.

OBJECTIVE: Epigallocatechin gallate (EGCG), the major antimicrobial tea polyphenol, has been reported to inhibit the growth of Candida albicans planktonic cells and enhance the antifungal activity of antimycotics. We hypothesised that synergism exists between EGCG and conventional antimycotics against biofilms of Candida species. DESIGN: The minimal inhibitory concentrations (MIC) of EGCG, miconazole, fluconazole and amphotericin B against planktonic cells and the sessile MIC (SMIC) against biofilms of Candida albicans, Candida parapsilosis, Candida tropicalis, Candida glabrata, Candida kefyr and Candida krusei were determined by a microdilution method. For assessment of biofilm metabolic activity, the 2,3-bis(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxanilide (XTT) assay was used. The interactions between EGCG and antimycotics were evaluated by checkerboard microdilution assay and determined by fractional inhibitory concentration index (FIC). RESULTS: Synergism was observed between EGCG and miconazole, fluconazole or amphotericin B against most test planktonic and biofilm cells of Candida species (FIC≤0.5). All biofilm cells were significantly more resistant to EGCG and antimycotics (20-3200 times higher) compared with their planktonic counterparts. CONCLUSIONS: We conclude that EGCG enhances the antifungal effects of miconazole, fluconazole and amphotericin B. Combined treatment with EGCG may lower the dosages of antimycotics, thus preventing adverse effects and the emergence of drug-resistant oral Candida species.

The antibiofilm activity of lingonberry flavonoids against oral pathogens is a case connected to residual complexity.

The antimicrobial activity of lingonberry (Vaccinium vitis-idaea L.) was evaluated against two oral pathogens, Streptococcus mutans and Fusobacterium nucleatum. Long-bed gel permeation chromatography (GPC; Sephadex LH-20) yielded purified flavonoids, with the most efficient minimum inhibitory concentrations (MICs) against planktonic cells in the anthocyanin and procyanidin primary fractions against F. nucleatum (63-125 µg/ml) and in the procyanidin rich fraction against S. mutans (16-31 µg/ml). The purified flavonol glycosides and procyanidins inhibited biofilm formation of S. mutans (MICs 16-31 µg/ml), while the corresponding reference compounds showed no activity. Secondary GPC purification yielded flavonol glycosides devoid of antibiofilm activity in the 50% MeOH fraction, while elution with 70% acetone recovered a brownish material with activity against S. mutans biofilm (MIC 8 µg/ml). Even after HPLC-PDA, NMR, and MALDI-TOF analyses, the structural identity of this material remained unknown, while its color and analytical characteristics appear to be consistent with flavonoid oxidation products.

The preventive effect of grape seed extract on artificial enamel caries progression in a microbial biofilm-induced caries model.

OBJECTIVES: The aim of this study was to evaluate the effect of grape seed extract (GSE) on enamel caries lesion formation in an in vitro Streptococcus mutans biofilm model. METHODS: Enamel fragments were prepared from bovine incisors and divided into six treatment groups (n=12): inoculated Brain Heart Infusion with 1% sucrose (BHIS), 1mg/mL GSE, 2mg/mL GSE, 3mg/mL GSE, 10ppm fluoride as NaF, and uninoculated BHIS. For biofilm formation, tooth fragments were incubated anaerobically in polystyrene 6-well tissue culture plates containing BHIS, the respective agents, and S. mutans (1×10(5)CFU/mL) for 24h at 37°C. Culture medium was replaced with fresh BHIS and respective agents daily over a 7-day period. Following caries lesion formation, lesion depth (LD) and relative optical density (ROD) were determined by polarized light microscopy (PLM) and confocal laser scanning microscopy (CLSM), respectively, to evaluate lesion progression. RESULTS: LDs of the 2mg/mL GSE group (122.86±13.41µm) and the 3mg/mL GSE group (111.92±11.39µm) were significantly smaller than those of the 1mg/mL GSE (198.33±17.70µm) and control groups (210.86±15.50µm) (p<0.05). Compared with the 2mg/mL and 3mg/mL groups, the control and 1mg/mL GSE groups showed significantly lower ROD values when depth was less than 200µm, indicating greater mineral loss. CONCLUSIONS: Dose-dependent GSE inhibits in vitro enamel caries formation due to its ability to suppress growth of S. mutans and the formation of biofilm. CLINICAL SIGNIfiCANCE: Grape seed extract may be a novel virulence-targeted natural antimicrobial agent for caries prevention.

Attachment of Porphyromonas gingivalis to corroded commercially pure titanium and titanium-aluminum-vanadium alloy.

BACKGROUND: Titanium dental material can become corroded because of electrochemical interaction in the oral environment. The corrosion process may result in surface modification. It was hypothesized that a titanium surface modified by corrosion may enhance the attachment of periodontal pathogens. This study evaluates the effects of corroded titanium surfaces on the attachment of Porphyromonas gingivalis. METHODS: Commercially pure titanium (cp-Ti) and titanium-aluminum-vanadium alloy (Ti-6Al-4V) disks were used. Disks were anodically polarized in a standard three-electrode setting in a simulated oral environment with artificial saliva at pH levels of 3.0, 6.5, or 9.0. Non-corroded disks were used as controls. Surface roughness was measured before and after corrosion. Disks were inoculated with P. gingivalis and incubated anaerobically at 37°C. After 6 hours, the disks with attached P. gingivalis were stained with crystal violet, and attachment was expressed based on dye absorption at optical density of 550 nm. All assays were performed independently three times in triplicate. Data were analyzed by two-way analysis of variance, the Tukey honestly significant difference test, t test, and Pearson's correlation test (α = 0.05). RESULTS: Both cp-Ti and Ti-6Al-4V alloy-corroded disks promoted significantly more bacterial attachment (11.02% and 41.78%, respectively; P <0.0001) than did the non-corroded controls. Significantly more (11.8%) P. gingivalis attached to the cp-Ti disks than to the Ti-6Al-4V alloy disks (P <0.05). No significant difference in P. gingivalis attachment was noted among the corroded groups for both cp-Ti and Ti-6Al-4V alloy (P >0.05). There was no significant correlation between surface roughness and P. gingivalis attachment. CONCLUSION: A higher degree of corrosion on the titanium surface may promote increased bacterial attachment by oral pathogens.

The effects of beverages on plaque acidogenicity after a sugary challenge.

BACKGROUND: Caries prevention traditionally has emphasized the restriction of cariogenic foods and beverages, but it has placed less emphasis on how the choice, combination and sequence of consumed foods and beverages may help reduce plaque acidogenicity. The authors conducted a study to examine whether whole milk, 100 percent apple juice or tap water affect dental plaque acidity in people after a sugary challenge. METHODS: Twenty adults participated in a randomized controlled crossover study. Participants consumed four combinations of foods: 20 grams of dry sugary Froot Loops (FL) (Kellogg's, Battle Creek, Mich.) cereal, 20 g of FL followed by 50 milliliters of milk (FL/milk), 20 g of FL followed by 50 mL of juice (FL/juice) and 20 g of FL followed by 50 mL of water (FL/water). The authors used a touch microelectrode to take plaque pH readings at the interproximal space just below the contact area between the maxillary premolars on both left and right sides at two and five minutes after FL consumption and at two to 30 minutes after milk, juice or water consumption. RESULTS: Consumption of FL plaque pH (standard deviation [SD]) was 5.83 (0.68) at 30 minutes, whereas plaque pH (SD) in the FL/milk group was 6.48 (0.30), which was significantly higher than that for FL/juice (5.83 [0.49]) or FL/water (6.02 [0.41]) (P < .005) at 35 minutes. CONCLUSION: Drinking milk after a sugary cereal challenge significantly reduced plaque pH drop due to the sugary challenge. PRACTICAL IMPLICATIONS: When discussing the cariogenicity of foods and beverages with patients, dentists and other health care professionals should emphasize that the order of ingesting sugary and nonsugary foods is important and may affect their oral health.

Physiological properties of Streptococcus mutans UA159 biofilm-detached cells.

Biofilm detachment is a physiologically regulated process that facilitates the release of cells to colonize new sites and cause infections. Streptococcus mutans is one of the major inhabitants of cariogenic dental plaque biofilm. This study tested the hypothesis that S. mutans biofilm-detached cells exhibit distinct physiological properties compared with their sessile and planktonic counterparts. Biofilm-detached cells showed a longer generation time of 2.85 h compared with planktonic cells (2.06 h), but had higher phosphotransferase activity for sucrose and mannose (P < 0.05). Compared with planktonic cells, they showed higher chlorhexidine (CHX) resistance and fourfold more adherent (P < 0.05). Increased mutacin IV production in biofilm-detached cells was noted by a larger inhibition zone against Streptococcus gordonii (31.07 ± 1.62 mm vs. 25.2 ± 1.74 mm by planktonic cells; P < 0.05). The expressions of genes associated with biofilm formation (gtfC and comDE) and mutacin (nlmA) were higher compared with planktonic cells (P < 0.05). In many properties, biofilm-detached cells shared similarity with sessile cells except for a higher phosphotransferase activity for sucrose, glucose, and mannose, increased resistance to CHX, and elevated expression of gtfC-, comDE-, and acidurity-related gene aptD (P < 0.05). Based on data obtained, the S. mutans biofilm-detached cells are partially distinct in various physiological properties compared with their planktonic and sessile counterparts.

An ORMOSIL-containing orthodontic acrylic resin with concomitant improvements in antimicrobial and fracture toughness properties.

Global increase in patients seeking orthodontic treatment creates a demand for the use of acrylic resins in removable appliances and retainers. Orthodontic removable appliance wearers have a higher risk of oral infections that are caused by the formation of bacterial and fungal biofilms on the appliance surface. Here, we present the synthetic route for an antibacterial and antifungal organically-modified silicate (ORMOSIL) that has multiple methacryloloxy functionalities attached to a siloxane backbone (quaternary ammonium methacryloxy silicate, or QAMS). By dissolving the water-insoluble, rubbery ORMOSIL in methyl methacrylate, QAMS may be copolymerized with polymethyl methacrylate, and covalently incorporated in the pressure-processed acrylic resin. The latter demonstrated a predominantly contact-killing effect on Streptococcus mutans ATCC 36558 and Actinomyces naselundii ATCC 12104 biofilms, while inhibiting adhesion of Candida albicans ATCC 90028 on the acrylic surface. Apart from its favorable antimicrobial activities, QAMS-containing acrylic resins exhibited decreased water wettability and improved toughness, without adversely affecting the flexural strength and modulus, water sorption and solubility, when compared with QAMS-free acrylic resin. The covalently bound, antimicrobial orthodontic acrylic resin with improved toughness represents advancement over other experimental antimicrobial acrylic resin formulations, in its potential to simultaneously prevent oral infections during appliance wear, and improve the fracture resistance of those appliances.

Efficacy of berberine, an antimicrobial plant alkaloid, as an endodontic irrigant against a mixed-culture biofilm in an in vitro tooth model.

INTRODUCTION: Berberine, a plant alkaloid isolated from many medicinal plants, has shown antimicrobial activity against selected oral pathogens. The purpose of this investigation was to evaluate the antimicrobial efficacy of berberine solution against selected endodontic pathogens using a multispecies biofilm tooth model. METHODS: The bacterial species used in the multispecies biofilm tooth model were Fusobacterium nucleatum, Enterococcus faecalis, and Prevotella intermedia. Extracted human anterior teeth were collected and standardized to a length of 14.0 mm. Teeth were cultured in Schaedler broth with the 3 test bacteria strains for 21 days and then randomly assigned to 6 treatment groups (ie, sterile saline, 5.25% NaOCl, 2% chlorhexidine [CHX], 1% CHX, 2 mg/mL berberine, and 1 mg/mL berberine plus 1% CHX). The teeth were instrumented to size 35/.06 and irrigated with 6 mL irrigant for 2 minutes. Surviving bacteria were sampled before and after instrumentation. Data were analyzed using analysis of variance (P < .05) followed by the Scheffé test. RESULTS: The minimal inhibitory concentration of berberine against F. nucleatum, P. intermedia, and E. faecalis was 31.25 µg/mL, 3.8 µg/mL, and 500 µg/mL, respectively. Instrumentation and irrigation resulted in 99% bacterial reduction in all groups. All tested solutions resulted in a statistically significant reduction in bacteria when compared with the saline control. When used alone, berberine (2 mg/mL) was less effective than the other test irrigants. However, when combined with 1% CHX, berberine (2 mg/mL) was comparable in bactericidal activity with 5.25% NaOCl, 2% CHX, and 1% CHX (Table 2). CONCLUSIONS: Berberine was more effective than saline as an endodontic irrigant against selected endodontic pathogens in vitro and, when combined with CHX, was comparable with NaOCl in its bactericidal efficacy.

Quaternary ammonium silane-functionalized, methacrylate resin composition with antimicrobial activities and self-repair potential.

The design of antimicrobial polymers to address healthcare issues and minimize environmental problems is an important endeavor with both fundamental and practical implications. Quaternary ammonium silane-functionalized methacrylate (QAMS) represents an example of antimicrobial macromonomers synthesized by a sol-gel chemical route; these compounds possess flexible Si-O-Si bonds. In present work, a partially hydrolyzed QAMS co-polymerized with 2,2-[4(2-hydroxy 3-methacryloxypropoxy)-phenyl]propane is introduced. This methacrylate resin was shown to possess desirable mechanical properties with both a high degree of conversion and minimal polymerization shrinkage. The kill-on-contact microbiocidal activities of this resin were demonstrated using single-species biofilms of Streptococcus mutans (ATCC 36558), Actinomyces naeslundii (ATCC 12104) and Candida albicans (ATCC 90028). Improved mechanical properties after hydration provided the proof-of-concept that QAMS-incorporated resin exhibits self-repair potential via water-induced condensation of organic modified silicate (ormosil) phases within the polymerized resin matrix.

Tea catechin epigallocatechin gallate inhibits Streptococcus mutans biofilm formation by suppressing gtf genes.

OBJECTIVE: The anti-cariogenic properties of tea have been suggested for decades. Tea polyphenols, especially epigallocatechin gallate (EGCG), have been shown to inhibit dental plaque accumulation, but the exact mechanisms are not clear at present. We hypothesise that EGCG suppresses gtf genes in Streptococcus mutans at the transcriptional level disrupting the initial attachment of S. mutans and thus the formation of mature biofilms. DESIGN: In this study, the effect of EGCG on the sucrose-dependent initial attachment of S. mutans UA159 in a chemically defined medium was monitored over 4 h using a chamber slide model. The effects of EGCG on the aggregation and gtf B, C, D gene expression of S. mutans UA159 were also examined. RESULTS: It was found that EGCG (7.8-31.25 µg/ml) exhibited dose-dependent inhibition of the initial attachment of S. mutans UA159. EGCG did not induce cellular aggregation of S. mutans UA159 at concentrations less than 78.125 µg/ml. Analysis of data obtained from real-time PCR showed that EGCG at sub-MIC level (15.6 µg/ml) significantly suppressed the gtf B, C, D genes of S. mutans UA159 compared with the non-treated control (p < 0.05). CONCLUSIONS: These findings suggest that EGCG may represent a novel, natural anti-plaque agent that inhibits the specific genes associated with bacterial biofilm formation without necessarily affecting the growth of oral bacteria.