Bacterial concentration and composition in liquid baby formula and a baby drink consumed with an artificial nipple

ObjectivesTo clarify the characteristics and growth of bacteria that may infiltrate liquid baby formula during feeding and after storage for more than 3 h, the transfer of oral bacteria through artificial nipples, and bacterial survival in liquid baby formula and a baby drink were examined immediately after drinking and after storage at 4 °C for 12 h and 24 h.MethodsThirteen human subjects (aged 19–24 years) were asked to drink approximately 50 mL of liquid baby formula and a baby drink, via the artificial nipple of a baby bottle. Samples of the remaining liquid after storage at 4 °C for 12 h and 24 h were inoculated onto blood agar plates and incubated anaerobically at 37 °C for 7 days. Genomic DNA was extracted from individual colonies, and the bacterial species were identified by 16S rRNA gene sequencing.ResultsThe mean concentrations of bacteria in the liquid baby formula were (2.6 ± 2.8) × 10⁴ and (4.1 ± 6.6) × 10⁴ colony-forming unit/mL after storage at 4 °C for 12 h and 24 h, respectively. Streptococcus (43.2%), Veillonella (9.3%), and Schaalia (8.2%) species were recovered from the remaining liquid baby formula after storage at 4 °C for 12 h. In contrast, no bacteria were detected in the remaining baby drink after storage at 37 °C for 24 h.ConclusionsThe levels of bacteria immediately after drinking and after storage at 4 °C for 12 h or 24 h were similar, suggesting that remaining liquid baby formula may be preserved safely in a refrigerator for more than 3 h.     

Profiling of the microbiota of breast milk before and after feeding with an artificial nipple

ObjectivesBreast milk is a valuable and useful source of nutrition; however, surplus milk is routinely discarded for hygiene reasons despite an unclear scientific basis. Here, we profiled the microbiota of expressed breast milk before and after feeding with an artificial nipple and examined the bacterial survival in breast milk stored at 4 °C.MethodsEleven mother–baby pairs were included in the study. Samples of expressed breast milk were collected before and after feeding with an artificial nipple and examined both immediately (0 h) and after storage for 3 and 12 h at 4 °C. Each sample was inoculated onto a blood agar plate and incubated anaerobically and aerobically at 37 °C. Genomic DNA was extracted from individual bacterial colonies, which were identified by 16S rRNA gene sequencing.ResultsBefore feeding, the bacterial counts at 0 and 12 h were (1.4 ± 1.6) × 10⁵ colony-forming units (CFU)/mL and (1.4 ± 0.6) × 10⁵ CFU/mL, respectively. Staphylococcus (47.7% and 41.9%, respectively), Cutibacterium (20.7% and 36.0%, respectively), and Streptococcus (16.1% and 6.6%, respectively) were identified among the samples. In contrast, after feeding, the bacterial counts at 0 and 12 h were (2.7 ± 1.7) × 10⁵ CFU/mL and (2.1 ± 2.5) × 10⁵ CFU/mL, respectively. Staphylococcus (30.1% and 37.4%, respectively), Cutibacterium (11.7% and 31.7%, respectively), and Streptococcus (41.5% and 25.2%, respectively), were identified among the samples.ConclusionsBacteria were present in the breast milk before feeding. Although the main component of the microbiota shifted from Staphylococcus to Streptococcus species after feeding, these results suggest that surplus expressed breast milk may be preserved safely in a refrigerator for at least 12 h after feeding with an artificial nipple.     

Microbiota profiles on the surface of non-woven fabric masks after wearing

This study aimed to characterize commensal microbiota on the skin before and after wearing masks, and to characterize the microbiota on the surface of used masks after 1 week of drying. From the 13 human subjects (age range, 19–26 years), mean bacterial concentrations of (6.1 ± 11.0) × 10⁵ and (1.0 ± 1.4) × 10⁶ colony-forming units (CFU)/mL were recovered from the skin of the buccal areas wiped with a sterile cotton swab before and after wearing non-woven fabric masks for 8 h, respectively. Furthermore (3.4 ± 4.9) × 10⁴ CFU/mL of bacteria were recovered from the mask surfaces. The bacteria contained in the masks, which consisted mainly of Cutibacterium acnes and Staphylococcus epidermidis/aureus, virtually disappeared after drying the masks indoors for 1 week.     

Efficacy of a mouthrinse based on hydroxyapatite to reduce initial bacterial colonisation in situ

ObjectiveThe present in situ - investigation aimed to specify the impact of pure hydroxyapatite microclusters on initial bioadhesion and bacterial colonization at the tooth surface.DesignPellicle formation was carried out in situ on bovine enamel slabs (9 subjects). After 1 min of pellicle formation rinses with 8 ml of hydroxyapatite (HA) microclusters (5%) in bidestilled water or chlorhexidine 0.2% were performed. As negative control no rinse was adopted. In situ biofilm formation was promoted by the intraoral slab exposure for 8 h overnight. Afterwards initial bacterial adhesion was quantified by DAPI staining and bacterial viability was determined in vivo/in vitro by live/dead-staining (BacLight). SEM analysis evaluated the efficacy of the mouthrinse to accumulate hydroxyapatite microclusters at the specimens’ surface and spit-out samples of the testsolution were investigated by TEM.ResultsCompared to the control (2.36 × 10⁶ ± 2.01 × 10⁶ bacteria/cm²), significantly reduced amounts of adherent bacteria were detected on specimens rinsed with chlorhexidine 0.2% (8.73 × 10⁴ ± 1.37 × 10⁵ bacteria/cm²) and likewise after rinses with the hydroxyapatite testsolution (2.08 × 10⁵ ± 2.85 × 10⁵ bacteria/cm², p < 0.001). No demonstrable effect of HA-particles on Streptococcus mutans viability could be shown. SEM analysis confirmed the temporary adsorption of hydroxyapatite microclusters at the tooth surface. Adhesive interactions of HA-particles with oral bacteria were shown by TEM.ConclusionHydroxyapatite microclusters reduced initial bacterial adhesion to enamel in situ considerably and could therefore sensibly supplement current approaches in dental prophylaxis.     

Efficacy of red propolis hydro-alcoholic extract in controlling Streptococcus mutans biofilm build-up and dental enamel demineralization

ObjectiveThe efficacy of a red propolis hydro-alcoholic extract (RP) in controlling Streptococcus mutans biofilm colonization was evaluated. The effect of RP on dental demineralization was also investigated.MethodsChemical composition was determined by High Performance Liquid Chromatography (HPLC). Minimum Inhibitory and Bactericidal Concentration (MIC and MBC, respectively) were investigated against Streptococcus mutans (ATCC 25175). The cytotoxic potential of 3% RP in oral fibroblasts was observed after 1 and 3 min. Bovine dental enamel blocks (N = 24) were used for S. mutans biofilm formation (48 h), simulating ‘feast or famine’ episodes. Blocks/biofilms were exposed 2×/day, for 3 days, to a cariogenic challenge with sucrose 10% (5 min) and treated (1 min) with: 0.85% saline solution (negative control), 0.12% Chlorhexidine (CHX, positive control for biofilm colonization), 0.05% Sodium Fluoride (NaF, positive control to avoid demineralization) and 3% RP. Biofilms were assessed for viability (CFU/mL), and to observe the concentration of soluble and insoluble extracellular polysaccharides (SEPS and IEPS). Dental demineralization was assessed by the percentage of surface hardness loss (%SHL) and through polarized light microscopy (PLM).ResultsThe RP presented 4.0 pH and ºBrix = 4.8. The p-coumaric acid (17.2 μg/mL) and luteolin (15.23 μg/mL) were the largest contents of phenolic acids and flavonoids, respectively. MIC and MBC of RP were 293 μg/mL and 1172 μg/mL, respectively. The 3% RP showed 43% of viably cells after 1 min. Lower number (p < 0.05) of viable bacteria (CFU/mL) was observed after CHX (1.8 × 10⁵) followed by RP (1.8 × 10⁷) treatments. The lowest concentration (μg/CFU) of SEPS (12.6) and IEPS (25.9) was observed in CHX (p < 0.05) followed by RP (17.1 and 54.3), and both differed from the negative control (34.4 and 63.9) (p < 0.05). Considering the %SHL, all groups differed statistically (p < 0.05) from the negative control (46.6%); but NaF (13.9%), CHX (20.1%) and RP (20.7%) did not differ among them (p > 0.05). After all treatments, suggestive areas of caries lesions were observed by PLM, which were lower for CHX and NaF.ConclusionThe 3% RP reduced S. mutans colonization, decreased concentration of extracellular polysaccharides and reduced dental enamel demineralization.     

Amazonian delicacy tucupi is as erosive as a cola-based soft drink

ObjectiveAcidic diets are advocated as main risk factor for tooth erosion, which could be prevented, or at least controlled, if patients were early advised. It is important to identify, hence, if possible dietary constituents regionally consumed on large scale, such as tucupi, a low-pH yellowish-green color and strong flavor delicacy made from the juice of a bitter cassava, may explain its occurrence in specific patient groups. This cross-over in situ/ex vivo study evaluated tucupi's ability to promote erosion of bovine enamel by assessing its percentage of surface microhardness change (%SMHC), taking a cola-based soft drink and human saliva as positive and negative controls.DesignFor three 7-days spaced out legs of 7 days each, nine volunteers wore palatal devices with three bovine enamel blocks, which were challenged with one of the following solutions: TUC—tucupi (n = 27); COL—cola-based soft drink (n = 27); SAL—saliva (n = 27). Erosive challenges were performed extra-orally (4×/day) by dropping TUC or COL at room temperature on specimens. After 5 min, palatal devices were replaced into the mouth. SAL permanently acted as the negative control while volunteers solely wore the device. One-way ANOVA followed by Tukey’s post-hoc tests (α = 0.05) were applied.ResultsTUC promoted an enamel %SMHC (−21.56 ± 10.08^a) similar than that promoted by COL (−18.19 ± 12.99^a; p = 0.275), which were both significantly higher than that promoted by SAL (−1.86 ± 13.65^b; p < 0.0001).ConclusionsBesides the most worldwide appreciated cola-based soft drink, the greatly consumed Amazonian delicacy tucupi can be considered a potential risk factor for tooth erosion.     

Antibacterial TAP-mimic electrospun polymer scaffold: effects on P. gingivalis-infected dentin biofilm

Objectives

This study sought to investigate, in vitro, the effects of a recently developed triple antibiotic paste (TAP)-mimic polymer nanofibrous scaffold against Porphyromonas gingivalis-infected dentin biofilm.

Materials and methods

Dentin specimens (4 × 4 × 1 mm³) were prepared from human canines. The specimens were sterilized, inoculated with P. gingivalis (ATCC 33277), and incubated for 1 week to allow for biofilm formation. Infected dentin specimens were exposed for 3 days to the following treatments: antibiotic-free polydioxanone scaffold (PDS, control), PDS + 25 wt% TAP [25 mg of each antibiotic (metronidazole, ciprofloxacin, and minocycline) per mL of the PDS polymer solution], or a saturated TAP-based solution (50 mg of each antibiotic per mL of saline solution). In order to serve as the negative control, infected dentin specimens were left untreated (bacteria only). To determine the antimicrobial efficacy of the TAP-mimic scaffold, a colony-forming unit (CFU) per milliliter (n = 10/group) measurement was performed. Furthermore, additional specimens (n = 2/group) were prepared to qualitatively study biofilm inhibition via scanning electron microscopy (SEM). Statistics were performed, and significance was set at the 5 % level.

Results

Both the TAP-mimic scaffold and the positive control (TAP solution) led to complete bacterial elimination, differing statistically (p < 0.05) from the negative control group (bacteria only). No statistical differences were observed for CFU per milliliter data between antibiotic-free scaffolds (2.7 log₁₀ CFU/mL) and the negative control (5.9 log₁₀ CFU/mL).

Conclusions

The obtained data revealed significant antimicrobial properties of the novel PDS-based TAP-mimic scaffold against an established P. gingivalis-infected dentin biofilm.

Clinical relevance

Collectively, the data suggest that the proposed nanofibrous scaffold might be used as an alternative to the advocated clinical gold standard (i.e., TAP) for intracanal disinfection prior to regenerative endodontics.

     

Effectiveness of 1-Visit and 2-Visit Endodontic Retreatment of Teeth with Persistent/Secondary Endodontic Infection: A Randomized Clinical Trial with 18 Months of Follow-up

IntroductionThis randomized clinical trial aimed to compare the effectiveness of endodontic retreatment of teeth with posttreatment apical periodontitis (PTAP) performed in 1 visit versus 2 visits on the reduction of cultivable bacteria (colony-forming units [CFUs]), lipopolysaccharides (LPSs), lipoteichoic acid (LTA), and the periapical lesion volume (mm³) after 18 months of follow-up.MethodsForty patients diagnosed with PTAP were selected and randomly divided into the following 2 groups: 1-visit retreatment and 2-visit retreatment with the placement of calcium hydroxide medication for 14 days. Cone-beam computed tomographic scans were performed at 2 stages: preoperatively and after 18 months of follow-up. Samples were collected before and after root canal procedures. A chromogenic limulus amebocyte lysate assay test was used to quantify endotoxins (EU/mL). LTA levels were measured using the enzyme-linked immunosorbent assay (pg/mL). Culture techniques were used to determine cultivable bacteria by counting the CFU (CFU/mL). The volume of the periapical lesions at the onset of the trial and at the 18-month posttreatment follow-up was obtained by the ITK-SNAP program (Cognitica, Philadelphia, PA).ResultsAll samples showed baseline values greater than those collected after the retreatment protocol for all investigated parameters. A higher bacterial load and lower LTA level were found in the 2-visit group after the retreatment protocol (P < .05), with no statistical differences between the groups regarding endotoxin levels and periapical lesion volume (mm³) at the 18-month follow-up analyzed by cone-beam computed tomographic imaging (P > .05).ConclusionsEndodontic retreatment in 1 or 2 visits exhibited equally favorable periapical healing at 18 months, with no statistically significant differences between groups.     

Effectiveness of curcumin against Enterococcus faecalis biofilm

Abstract

Objectives. To evaluate the antimicrobial efficacy of curcumin against Enterococcus faecalis bio?lm formed on tooth substrate in vitro. Sodium hypochlorite (NaOCl) and chlorhexidine (CHX) served as standards for comparison. Materials and methods. Biofilms of E.faecalis were formed on instrumented, extracted human teeth (n = 96). At the end of the 2nd day, 2nd and 8th weeks, specimens were treated for 30 min with one of the test solutions or saline (control) and the surviving colony-forming units (CFU/mL) was recorded. Results were analyzed by Kruskal-Wallis test and Dunnet test for pair-wise comparison with Bonferroni correction (p = 0.05). Results. Only NaOCl showed complete eradication of bacteria at all time periods. In the 2-day and 2nd week biofilms, curcumin and NaOCl showed complete inhibition, which was significantly lower than the CFU recovered in the CHX and saline groups (p < 0.05). In 8 week biofilms, samples treated with curcumin showed 553 ± 137.6 CFU/mL, which was significantly higher than NaOCl (0 CFU/mL), but significantly lower than CHX (2551 ± 129.8) and saline control (1.42 × 1011 ± 2.12 × 1010; p < 0.05). Conclusions. Sodium hypochlorite (3%) showed maximum antibacterial activity against E.faecalis bio?lm formed on the tooth substrate, followed by curcumin and CHX. Considering the potential for undesirable properties of NaOCl, the use of herbal alternatives in endodontics might prove to be advantageous.     

Effectiveness of calcium hydroxide-based intracanal medication on infectious/inflammatory contents in teeth with post-treatment apical periodontitis

Objective

The objective of this work was to investigate in vivo the effects of calcium hydroxide-based intracanal medication (ICM) on the levels of bacteria, pro-inflammatory cytokines (PICs), and matrix metalloproteinases (MMPs) in root canals and periradicular tissues of teeth with failure of the root canal treatment and apical periodontitis.

Materials and methods

Twenty infected root canals of single-rooted teeth were randomly assigned into two groups according to the irrigant used for chemomechanical preparation (CMP) (n = 10 per group): G1 – 2% chlorhexidine (CHX) gel and G2 – 6% sodium hypochlorite (NaOCl). Root canal contents were taken by using paper points before CMP (S1) and after 30 days of calcium hydroxide-based ICM (S2). Microbial reduction was calculated by means of colony-forming unit count (CFU/mL), with PICs and MMPs (pg/mL) being measured by using enzyme-linked immunosorbent assay (ELISA).

Results

Culturable bacteria (101.2 ± 79.2), PICs (IL-1β 1.2 ± 0.4 and TNF-α 8.8 ± 4.7), MMP-2 (803.7 ± 96.4), MMP-3 (453.9 ± 229.3), MMP-8 (245.9 ± 122.4), MMP-9 (129.4 ± 29.6), and MMP-13 (70.8 ± 12.8) were present in all S1 samples. After 30 days of ICM (S2), a 99.5% microbial reduction was observed, together with a significant reduction of PICs in all groups. Overall, it was observed a decrease in the levels of MMPs (S2), except MMP-13, which was found in increased levels after ICM (P < .05), independently of the groups.

Conclusions

Calcium hydroxide-based intracanal medications have had a positive effect on the microbial reduction by decreasing the levels of PICs and MMPs. Both auxiliary chemical substances (i.e., 2% CHX and 6% NaOCl) presented similar effects when calcium hydroxide was used as intracanal medication.

Clinical relevance

Teeth with failure of the root canal treatment and apical periodontitis, and consequently with high levels of bacteria, PIC, and MMP, may present a better prognosis after a 30 days of a calcium hydroxide-based ICM.

     

Assessment of Root Canal Sealers Loaded with Drug-Silica Coassembled Particles Using an In Vitro Tooth Model

IntroductionThe purpose of this study was to assess the antimicrobial activity of root canal sealers modified with novel highly loaded antimicrobial drug-silica coassembled particles (DSPs) on Enterococcus faecalis–infected root canal dentin.MethodsDSPs were synthesized through coassembly of silica and octenidine dihydrochloride (OCT) surfactant drug (35% w/w OCT). DSPs (1% wt of the total mass of the sealer) were mixed homogenously with either epoxy resin sealer (AH Plus [AH]; Dentsply Sirona, Tulsa, OK) or calcium silicate–based sealer (EndoSequence BC Sealer [BC]; Brasseler, Savannah, GA). To assess the antimicrobial activity of DSP-loaded sealers, the apical third of single-rooted teeth was obtained and infected with E. faecalis for 3 weeks followed by the application of experimental (DSP-loaded) sealers or corresponding controls for up to 28 days. Microbiological analysis and laser scanning confocal and scanning electron microscopy were used to determine the colony-forming unit (CFU)/mL, the percentage of live bacteria, and the intratubular bacterial and sealer penetrations. Factorial analysis of variance and Tukey post hoc tests were used to assess the antimicrobial effect of DSPs on different sealers.ResultsAll experimental groups showed significant reductions in CFUs at all-time points compared with positive controls (P < .05). The addition of DSPs to BC significantly reduced the CFUs (2.11 ± 0.13, 2.22 ± 0.19, and 2.25 ± 0.17 at 1, 7, and 28 days, respectively) compared with the unmodified sealer (3.21 ± 0.11, 4.3 ± 0.15, and 4.2 ± 0.2 at 0, 7, and 28 days). DSPs enhanced the antimicrobial performance of AH only at 1 day (4.21 ± 0.17 vs 5.19 ± 0.12, P < .05). AH and AH + DSPs showed higher bacterial viability compared with BC and BC + DSPs at all incubation periods (P < .05).ConclusionsLoading endodontic sealers with DSPs had a material-dependent effect on the antimicrobial properties and could reduce the incidence of secondary infections.     

Efficacy of tissue engineered bone grafts containing mesenchymal stromal cells for cleft alveolar osteoplasty in a rat model

The development of sufficient tissue engineered bone grafts for alveolar cleft osteoplasty could reduce the necessity of autogenous bone grafts and its donor site morbidity. The aim of the study was to evaluate tissue engineered bone grafts in an artificially created bone defect.Bone grafts were created in vitro colonizing a synthetic hydroxyapatite–tricalciumphosphate scaffold (BONITmatrix^®) with either undifferentiated mesenchymal stromal cells (group 1) or osteogenic differentiated mesenchymal stromal cells (group 2). Cells were multiplied from bone marrow of donor rats. Unmodified scaffolds (group 3) and the tissue engineered bone grafts were inserted into artificial maxillary defects of 54 Lewis rats. In 18 animals the defects remained unfilled (control). After one, three and six weeks the rats were sacrificed. The defect was evaluated radiologically and histologically with regard to the remaining defect volume and diameter. Statistical analysis followed.The bone grafts led to a specific bone formation at the defect margin. No complete reunion of any defect was observed within the healing time. After six weeks, the remaining defect volume was 6.86 ± 3.21 mm³ (control), 4.08 ± 1.36 mm³ (group 1), 5.00 ± 0.84 mm³ (group 2) 5.50 ± 1.05 mm³ (group 3). The remaining defect diameter measured 2.63 ± 0.52 mm (control), 2.39 ± 0.23 mm (group 1), 2.53 ± 0.22 mm (group 2) and 2.70 ± 0.66 mm (group 3). In all experimental groups the defect volume and diameter decreased over time, which was significant for group 1 (p = 0.014), group 2 (p = 0.025) and group 3 (p = 0.048). The defect volume and width was significantly reduced for bone grafts containing undifferentiated cells compared to control (p = 0.035) or scaffolds only (p = 0.05).ConclusionTissue engineered bone grafts induce a pronounced bone formation in artificial bone defects compared to unfilled controls or scaffolds only.     

Efficacy of doxycycline release collagen membrane on surgically created and contaminated defects in rat tibiae: A histopathological and microbiological study

BackgroundThe effects of systemic antibiotics on controlling infective pathogens after guided bone regeneration(GBR) procedures especially in membrane exposures are limited. However, local administrations of antibiotics are rare in GBR techniques.AimThe aim of this study was to investigate the osteogenesis potential and the antibacterial effect of a doxycycline releasing collagen membrane in surgically created and contaminated defects in rat tibiae.Material and methodsDefects were created in 20 rats that were randomly divided in to two groups: control group (defect contaminated by Porphyromonas gingivalis, filled with bone graft and covered by collagen membrane); test group (defect contaminated by P. gingivalis filled with bone graft and covered by collagen membrane containing 1 mg/cm² doxycycline. Animals were sacrificed post surgically on the 14th day for microbiologic evaluation and on the 28th day for histopathological evaluation.ResultsThe degree of osteogenesis in the test group was seen to be significantly higher than control group (p: 0.011; p < 0.05). Furthermore in test group, no bacterial growth was observed. The bacteria counts were determined between 1 × 104 and 268 × 104 CFU/g with a median of 1.32 × 104 for control group.ConclusionsWithin the limitations of this study, the results of the present study suggests that the use of a doxycycline releasing membrane has a positive effect on contaminated GBR procedures for limiting P. gingivalis infections leading to bone formation following GBR procedures in a rat model.     

Influence of a Brazilian wild green propolis on the enamel mineral loss and Streptococcus mutans’ count in dental biofilm

ObjectiveThis study investigated the anti-demineralizing and antibacterial effects of a propolis ethanolic extract (EEP) against Streptococcus mutans dental biofilm.DesignBlocks of sound bovine enamel (n = 24) were fixed on polystyrene plates. S. mutans inoculum (ATCC 25175) and culture media were added (48 h–37 °C) to form biofilm. Blocks with biofilm received daily treatment (30 μL/1 min), for 5 days, as following: G1 (EEP 33.3%); G2 (chlorhexidine digluconate 0.12%); G3 (ethanol 80%); and G4 (Milli-Q water). G5 and G6 were blocks without biofilm that received only EEP and Milli-Q water, respectively. Final surface hardness was evaluated and the percentage of hardness loss (%HL) was calculated. The EEP extract pH and total solids were determined. S. mutans count was expressed by log₁₀ scale of Colony-Forming Units (CFU/mL). One way ANOVA was used to compare results which differed at a 95% significance level.ResultsG2 presented the lowest average %HL value (68.44% ± 12.98) (p = 0.010), while G4 presented the highest (90.49% ± 5.38%HL) (p = 0.007). G1 showed %HL (84.41% ± 2.77) similar to G3 (87.80% ± 6.89) (p = 0.477). Groups G5 and G6 presented %HL = 16.11% ± 7.92 and 20.55% ± 10.65; respectively (p = 0.952). G1 and G4 differed as regards to S. mutans count: 7.26 ± 0.08 and 8.29 ± 0.17 CFU/mL, respectively (p = 0.001). The lowest bacterial count was observed in chlorhexidine group (G2 = 6.79 ± 0.10 CFU/mL) (p = 0.043). There was no difference between S. mutans count of G3 and G4 (p = 0.435). The EEP showed pH 4.8 and total soluble solids content = 25.9 Brix.ConclusionThe EEP seems to be a potent antibacterial substance against S. mutans dental biofilm, but presented no inhibitory action on the de-remineralization of caries process.     

In vitro evaluation of the erosive potential of viscosity-modified soft acidic drinks on enamel

Objective

The objective of this in vitro study was to investigate the effect of viscosity-modified soft acidic drinks on enamel erosion.

Materials and methods

A total of 108 bovine enamel samples (??=?3 mm) were embedded in acrylic resin and allocated into six groups (n?=?18). Soft acidic drinks (orange juice, Coca-Cola, Sprite) were used both in their regular forms and at a kinetic viscositiy of 5 mm²/s, which was adjusted by adding hydroxypropyl cellulose. All solutions were pumped over the enamel surface from a reservoir with a drop rate of 3 ml/min. Each specimen was eroded for 10 min at 20 °C. Erosion of enamel surfaces was measured using profilometry. Data were analyzed using independent t tests and one-way ANOVAs (p?<?0.05).

Results

Enamel loss was significantly higher for the regular (Coca-Cola, 5.60?±?1.04 μm; Sprite, 5.49?±?0.94 μm; orange juice, 1.35?±?0.4 μm) than for the viscosity-modified drinks (Coca-Cola, 4.90?±?0.34 μm; Sprite, 4.46?±?0.39 μm; orange juice, 1.10?±?0.22 μm).

Conclusion

For both regular and viscosity-modified forms, Coca-Cola and Sprite caused higher enamel loss than orange juice. Increasing the viscosity of acidic soft drinks to 5 mm²/s reduced enamel erosion by 12.6–18.7 %.

Clinical relevance

The erosive potential of soft acidic drinks is not only dependent on various chemical properties but also on the viscosity of the acidic solution and can be reduced by viscosity modification.     

Dual function of quercetin as an MMP inhibitor and crosslinker in preventing dentin erosion and abrasion: An in situ/in vivo study

ObjectiveThe aim of the present study was to evaluate the in situ/in vivo effect of quercetin on dentin erosion and abrasion.MethodsHuman dentin blocks (2 × 2 × 2 mm) were embedded and assigned to 6 groups: 75 μg/mL, 150 μg/mL and 300 μg/mL quercetin (Q75, Q150, Q300); 120 μg/mL chlorhexidine (CHX, positive control); and deionized water and ethanol (the negative controls). The specimens were treated with the respective solutions for 2 min and then subjected to in situ/in vivo erosive/abrasive challenge for 7 d as follows: in vivo erosion 4 times a day and then in vivo toothbrush abrasion after the first and last erosive challenges of each day. Dentin loss was assessed by profilometry. An additional dentin specimen was used to evaluate the penetration depth of quercetin into dentin by tracking the spatial distribution of its characteristic Raman peak. Moreover, dentin blocks (7 × 1.7 × 0.7 mm) were used to detect the impact of quercetin on dentin-derived matrix metalloproteinase (MMP) inhibition by in situ zymography, and the inhibition percentage (%) was calculated. Additionally, the potential collagen crosslinking interactions with quercetin were detected by Raman spectroscopy, and the crosslinking degree was determined with a ninhydrin assay. Fully demineralized dentin beams (0.5 × 0.5 × 10 mm) were used to evaluate the impact of quercetin on the mechanical properties of dentin collagen fibre by the ultimate micro-tensile strength test (μUTS). The data were analysed by one-way analysis of variance and Tukey’s test (α = 0.05).ResultsCompared to the negative controls, all treatment solutions significantly reduced dentin loss. The dentin loss of Q150 and Q300 was significantly less than that of CHX (all P < 0.05). The amount of quercetin decreased with increasing dentin depth, and the maximum penetration depth was approximately 25–30 µm. In situ zymography showed that quercetin significantly inhibited the activities of dentin-derived MMPs. The inhibitory percentages of Q75 and Q150 were significantly lower than that of CHX (all P < 0.05), but no significant difference was found between Q300 and CHX (P = 0.58). The collagen crosslinking interactions with quercetin primarily involved hydrogen bonding and the degree of crosslinking increased in a concentration-dependent manner. Statistically significant increases in μUTS values were observed for demineralized dentin beams after quercetin treatment compared with those of the control treatments (all P < 0.05).SignificanceThis study provides the first direct evidence that quercetin could penetrate approximately 25–30 µm into dentin and further prevent dentin erosion and abrasion by inhibiting dentin-derived MMP activity as well as crosslinking collagen of the demineralized organic matrix.     

Denaturing gradient gel electrophoresis profiles of bacteria from the saliva of twenty four different individuals form clusters that showed no relationship to the yeasts present

ObjectivesThe aim was to investigate the relationship between groups of bacteria identified by cluster analysis of the DGGE fingerprints and the amounts and diversity of yeast present.MethodsBacterial and yeast populations in saliva samples from 24 adults were analysed using denaturing gradient gel electrophoresis (DGGE) of the bacteria present and by yeast culture.ResultsEubacterial DGGE banding patterns showed considerable variation between individuals. Seventy one different amplicon bands were detected, the band number per saliva sample ranged from 21 to 39 (mean ± SD = 29.3 ± 4.9). Cluster and principal component analysis of the bacterial DGGE patterns yielded three major clusters containing 20 of the samples. Seventeen of the 24 (71%) saliva samples were yeast positive with concentrations up to 10³ cfu/mL. Candida albicans was the predominant species in saliva samples although six other yeast species, including Candida dubliniensis, Candida tropicalis, Candida krusei, Candida guilliermondii, Candida rugosa and Saccharomyces cerevisiae, were identified. The presence, concentration, and species of yeast in samples showed no clear relationship to the bacterial clusters.ConclusionDespite indications of in vitro bacteria-yeast interactions, there was a lack of association between the presence, identity and diversity of yeasts and the bacterial DGGE fingerprint clusters in saliva. This suggests significant ecological individual-specificity of these associations in highly complex in vivo oral biofilm systems under normal oral conditions.     

Elevation of white blood cells and platelet counts in patients having chronic periodontitis

Background and aimMany risk factors that might contribute to the pathogenesis of atherosclerosis have been proposed, including chronic inflammation and infection. Furthermore, systemic inflammatory responses to periodontal bacteria have been suggested as a pathogenetic link between periodontal disease and atherosclerosis. The purpose of this study was to estimate the white blood cell (WBC) and platelet counts in chronic periodontitis patients.Materials and methodsFifty patients with chronic periodontitis and 50 patients with healthy periodontium were included in this study. Oral hygiene status, pocket probing depth (PPD) and clinical attachment level (CAL) were measured. During clinical evaluation, venous blood samples were taken to analyze the WBC and platelet counts. Statistical analysis was utilized to compare differences across groups.ResultsPeriodontitis patients demonstrated a significantly higher WBC count (7.22 ± 1.42 × 10⁹ cells/L) than that of control patients (5.64 ± 1.56 × 10⁹ cells/L; P < 0.001). The platelet count of patients with chronic periodontitis (290.73 ± 56.56 × 10⁹ cells/L) was also significantly higher compared to the healthy group (223.37 ± 50.27 × 10⁹ cells/L; P < 0.001).ConclusionLevels of WBCs and platelets are elevated in periodontitis patients compared to healthy controls.     

Advanced co-culture model: Soft tissue cell and bacteria interactions at the transgingival dental implant interface

ObjectivesTo better simulate and understand the clinical situation in which tissue cells and bacteria compete for settlement on an implant surface, the aim was to develop an improved transgingival co-culture model.MethodsFor this model human gingival fibroblasts (HGF) were seeded on different titanium surfaces in the presence of the early colonizer Streptococcus gordonii or mixed oral bacteria. Subsequently adhesion and viability of HGF cells was analyzed.ResultsSimultaneous co-culture showed no decrease in the viability of HGF cells at early stages compared to the control group. However, a moderate impact on HGF viability (76 ± 23 %) was observed after 4 h of co-culture, which then significantly decreased after 5 h (21 ± 2 %) of co-cultivation, resulting in cell death and detachment from the surface. Further experiments including saliva pre-treatment of smooth and structured titanium surfaces with Streptococcus gordonii or mixed oral bacteria suggested a cell-protective property of saliva.SignificanceOur study revealed that during simultaneous co-culture of cells and bacteria, which resembles the clinical situation the closest, the viability of gingival cells is considerably high in the early phase, suggesting that increasing initial cell adhesion rather than antibacterial functionality is a major goal and a relevant aspect in the development and testing of transgingival implant and abutment surface modifications.