Efficacy of mouthrinses with bovine milk and milk protein isolates to accumulate casein in the in situ pellicle.

OBJECTIVES: The adsorption of bovine milk caseins on the tooth surface might have a positive impact on the prevention of dental diseases. Therefore, the present study aimed to investigate the efficacy of mouthrinses with different types of bovine milk and milk protein isolates to accumulate caseins in the pellicle. MATERIALS/METHODS: An indirect enzyme-linked immunosorbent assay (ELISA) was established to quantify the amount of caseins adsorbed into the in situ pellicle. In situ pellicle samples were collected from 2 volunteers on ceramic specimens (A = 8 cm2). After 10 min of pellicle formation, different types of bovine milk, 3% micellar casein in synthetic milk ultrafiltrate (SMUF) or 3% non-micellar caseinate in SMUF, were used as mouthrinses for 10 min. The pellicle material was harvested after 30 min in situ and examined for caseins by the indirect ELISA. Selected pellicle samples were subjected to TEM analysis. RESULTS: All mouthrinses accumulated caseins in the in situ pellicle (2.0 ± 0.7-20 ± 1.7 µg/ml) that, under native conditions, expressed no casein signal. Micellar protein association increased the adsorption of casein into the pellicle. Milk homogenization also had an influence on the casein accumulation in the pellicle. TEM analysis confirmed the integration of micellar casein into the pellicle. CONCLUSION: The mouthrinses altered the protein composition and the ultrastructure of the in situ pellicle to a different extent: bovine milk with 3.8% fat content and 3% micellar casein in SMUF being particularly effective. CLINICAL RELEVANCE: The study provides interesting perspectives for innovative prevention strategies in dentistry.

Shear bond strength of different types of adhesive systems to dentin and enamel of deciduous teeth in vitro.

OBJECTIVES: The present study aimed to evaluate the suitability of self-etch adhesives for restoration of deciduous teeth compared with etch and rinse approaches. MATERIALS AND METHODS: One hundred twenty primary teeth were divided into five groups, each being assigned to an adhesive system. Self-etch adhesives XenoV (XV) and Clearfil S(3) Bond (CB), Prime&Bond NT with (PBE)/without preliminary etching (PBN), and Optibond FL (OBFL) as an etch and rinse system were included. Enamel and dentin specimens were prepared (n = 36/group), adhesives applied, and compomer cylinders polymerized. After 24-h storage in 37 °C distilled water and thermo-cycling (1440 cycles, 5/55 °C, 27 s), shear bond tests and fracture mode classification based on SEM investigation were performed. Statistical analysis involved ANOVA and Scheffé procedure with Bonferroni-Holm correction (p ≤ 0.005). RESULTS: High shear bond strengths to primary enamel were determined for PBE (mean [M] = 22.48 ± 7.7 MPa) > OBFL (M = 19.06 ± 5.62 MPa) > CB (M = 17.6 ± 6.55 MPa), and XV (M = 16.85 ± 5.38 MPa) and PBN (M = 8.26 ± 4.46 MPa) formed significantly less reliable enamel-resin interfaces (p ≤ 0.005). PBE generated the highest bond strength on primary dentin (M = 21.97 ± 8.02 MPa); significantly lower values were measured for XV (M = 13.44 ± 5.43 MPa) and OBFL (M = 12.92 ± 4.31 MPa) (p ≤ 0.005). CONCLUSIONS: Adhesives requiring preliminary etching ensure optimal bond strength to primary enamel. If separate etching is to be avoided, selected self-etch adhesives obtain acceptable shear bond values on primary enamel and dentin. CLINICAL RELEVANCE: The treatment of pediatric patients presents a great challenge in dental practice, and optimization of treatment processes is important.

Influence of Calcium Phosphate and Apatite Containing Products on Enamel Erosion.

For the purpose of erosion prevention the present study aimed to compare the efficacy of two biomimetic products and a fluoride solution to optimize the protective properties of the pellicle. After 1 min of in situ pellicle formation on bovine enamel slabs, 8 subjects adopted CPP-ACP (GC Tooth Mousse), a mouthwash with hydroxyapatite microclusters (Biorepair), or a fluoride based mouthwash (elmex Kariesschutz) for 1 min each. Afterwards, samples were exposed in the oral cavity for 28 min. Native enamel slabs and slabs exposed to the oral cavity for 30 min without any rinse served as controls. After oral exposure, slabs were incubated in HCl (pH values 2, 2.3, and 3) for 120 s and kinetics of calcium and phosphate release were measured photometrically; representative samples were evaluated by SEM and TEM. The physiological pellicle reduced demineralization at all pH values; the protective effect was enhanced by fluoride. The biomimetic materials also reduced ion release but their effect was less pronounced. SEM indicated no layer formation after use of the different products. However, TEM confirmed the potential accumulation of mineral components at the pellicle surface. The tested products improve the protective properties of the in situ pellicle but not as effectively as fluorides.

Changes in Candida spp., mutans streptococci and lactobacilli following treatment of early childhood caries: a 1-year follow-up.

Early childhood caries (ECC) is closely related to high numbers of mutans streptococci, lactobacilli and Candida albicans. Oral colonization of these microorganisms was monitored in a prospective clinical study in order to investigate the effect of comprehensive treatment under general anesthesia and the sustainability of microbial changes. Saliva samples were collected from 50 healthy infants with ECC before and in regular intervals up to 12 months after treatment. Microorganisms were detected by cultivation on selective agars (CRT® bacteria and Sabouraud/CandiSelect™) and scored. Additionally, plaque on upper front teeth and the dmft were recorded. Parents were repeatedly interviewed regarding the children's diet and oral hygiene, accompanied by corresponding advice. Plaque frequency and the numbers of mutans streptococci, lactobacilli and yeasts were significantly reduced as a result of treatment (p < 0.0001, Wilcoxon test). Nevertheless, this effect was not permanent. An ordinal regression model on the follow-up period revealed that the odds for bacteria and yeasts to reach a higher score increased linearly over time (p < 0.01) with an odds ratio of 2.244 per year. One third (34%) of the children developed new dentinal lesions within 1 year postoperatively. High scores of lactobacilli before treatment predicted caries relapse (p < 0.05). Nutritional and oral hygiene habits changed only slightly despite advising. Elimination and restoration of ECC lesions under general anesthesia proved to be an effective procedure in reducing cariogenic bacteria and yeasts. A satisfactory and sustainable success, however, could be achieved neither regarding microbiologic parameters nor with respect to the relapse rate. More suitable strategies are needed.

Impact of pH-adjusted fluoride and stannous solutions on the protective properties on the pellicle layer in vitro and in situ.

This study evaluates the ideal pH for anti-erosion and anti-adherent efficacy of fluoride and stannous solutions (sodium fluoride (SF), amine fluoride (AF), sodium monofluorophosphate (SMFP), stannous fluoride (SnF2) with 500 ppm fluoride concentration each and stannous chloride (SnCl2, 1563 ppm stannous)). In vitro, solutions were tested at pH 4.5 and 5.5. The main in situ experiments were carried out at the pH of 4.5: For pellicle formation 6 volunteers wore bovine enamel slabs intraorally for 1 min, rinsed with 8 ml solution for 1 min and continued for up to 30 min/8 h. Physiological pellicle samples served as controls. After incubation in HCl (2.0, 2.3) for 2 min mineral release was determined photometrically. Bacterial counts on 8 h biofilms were determined by fluorescence microscopy (BacLight™ and DAPI with Concanavalin A). Modification of the pellicle ultrastructure was examined by TEM. Statistical analysis was performed using Kruskal-Wallis and Mann-Whitney-U tests with Bonferroni-correction (p < 0.05). SnF2 showed a significant erosion protection. AF, SnF2, and SnCl2 were most anti-adherent. SnF2 and SnCl2 caused a pronounced basal pellicle with stannous precipitates. Compared to other fluoride monosubstances, stannous ions offer greater protection against erosive acidic attacks. Stannous ions act as crucial co-factor in this process.

Effect of conventional mouthrinses on initial bioadhesion to enamel and dentin in situ.

AIM: The study aimed to investigate the effect of a customary fluoride solution, containing sodium fluoride and amine fluoride, on initial biofilm formation on enamel and dentin in situ compared directly to chlorhexidine. METHODS: Bovine enamel and dentin specimens were mounted on maxillary splints carried by 9 subjects. After 1 min of pellicle formation, rinses with tap water (control), chlorhexidine (meridol med CHX 0.2%, GABA) and a fluoride mouthrinse (elmex, GABA) were performed for 1 min. Subsequently, the slabs were carried for another 8 h. The adherent bacteria were determined by DAPI staining, live-dead staining and determination of colony-forming units after desorption; glucan formation was visualized with concanavalin A. Additionally, energy-dispersive X-ray spectroscopy (EDX) analysis of the in situ biofilm layers was conducted, and contact angle measurements were performed. Statistical evaluation was performed by means of the Kruskal-Wallis test followed by the Mann-Whitney U test (p < 0.05). RESULTS: In the control group, significantly higher amounts of adherent bacteria were detected on dentin (4.8 × 10(6) ± 5.4 × 10(6) bacteria/cm(2)) than on enamel (1.2 × 10(6) ± 1.5 × 10(6) bacteria/cm(2), DAPI). Chlorhexidine significantly reduced the amount of adherent bacteria (dentin: 2.8 × 10(5) ± 3.4 × 10(5) bacteria/cm(2); enamel: 4.2 × 10(5) ± 8.7 × 10(5) bacteria/cm(2)). Rinses with the fluoride solution also significantly reduced bacterial adherence to dentin (8.1 × 10(5) ± 1.5 × 10(6) bacteria/cm(2)). Fluoride could not be detected by EDX analysis of the biofilms. Fluoride mouthrinsing did not influence the wettability of the pellicle-covered enamel surface. CONCLUSION: In addition to the reduction of demineralization and antibacterial effects, fluorides inhibit initial biofilm formation on dental hard tissues considerably, especially on dentin.

Influence of a mouthwash containing hydroxyapatite microclusters on bacterial adherence in situ.

OBJECTIVE: The aim of the present study was to investigate the efficacy of a new preparation in dental prophylaxis containing zinc-carbonate hydroxyapatite microclusters (Biorepair) for oral biofilm management. METHODS AND MATERIALS: Initial biofilm formation was carried out in situ with bovine enamel slabs fixed to individual upper jaw splints worn by six subjects. Rinses with the customary preparation as well as with subfractions (hydroxyapatite microclusters in saline solution; liquid phase without particles) were adopted for 1 min in situ after 1 min of pellicle formation, and the bacterial colonization was recorded after 6 h and 12 h, respectively. Rinses with chlorhexidine served as a reference. The adherent microorganisms were quantified and visualized using DAPI staining and live-dead staining (BacLight). Furthermore, the effects on Streptococcus mutans bacteria were tested in vitro (BacLight). RESULTS: Application of the customary preparation and of the separate components distinctly reduced the initial bacterial colonization of the enamel surface in situ as visualized and quantified with all techniques. After 12 h, 1.3 × 10(7) ± 2.0 × 10(7) bacteria/cm² were detected on unrinsed control samples with DAPI staining; 2.4 × 10(6) ± 3.3 × 10(6) after application of Biorepair (12 h after CHX-rinse; 1.3 × 10(5) ± 9.2 × 10(4)). Also, pure hydroxyapatite microclusters in saline solution (2.1 × 10(6) ± 3.0 × 10(6)) as well as the liquid phase without particles (5.1 × 10(5) ± 3.3 × 10(5)) reduced the amount of adherent bacteria. Furthermore, antimicrobial effects on S. mutans were observed in vitro. CONCLUSION: The preparation is an effective compound for biofilm management in the oral cavity due to antiadherent and antibacterial effects. CLINICAL RELEVANCE: The tested mouthrinse seems to be a reasonable amendment for dental prophylaxis.

Lipids in preventive dentistry.

OBJECTIVES: There is still a great demand for the improvement of oral prophylaxis methods. One repeatedly described approach is rinsing with edible oils. The aim of the present review paper was to analyze the role of lipids in bioadhesion and preventive dentistry. MATERIALS AND METHODS: Despite limited sound scientific data, extensive literature search was performed to illustrate possible effects of lipids in the oral cavity. RESULTS: It is to be assumed that lipophilic components modulate the process of bioadhesion to the oral hard tissues as well as the composition and ultrastructure of the initial oral biofilm or the pellicle, respectively. Thereby, lipids could add hydrophobic characteristics to the tooth surface hampering bacterial colonization and eventually decreasing caries susceptibility. Also, a lipid-enriched pellicle might be more resistant in case of acid exposure and could therefore reduce the erosive mineral loss. Furthermore, anti-inflammatory effects on the oral soft tissues were described. However, there is only limited evidence for these beneficial impacts. Neither the lipid composition of saliva and pellicle nor the interactions of lipids with the initial oral biofilm and the pellicle layer have been investigated adequately until now. CONCLUSION: Edible oils might qualify as mild supplements to conventional strategies for the prevention of caries, erosion, and periodontal diseases but further research is necessary. CLINICAL RELEVANCE: Against the background of current scientific and empirical knowledge, edible oils might be used as oral hygiene supplements but a decisive benefit for the oral health status is questionable.

Comparison of different live/dead stainings for detection and quantification of adherent microorganisms in the initial oral biofilm.

OBJECTIVES: The aim of the present study was to investigate different fluorescence-based, two-color viability assays for visualization and quantification of initial bacterial adherence and to establish reliable alternatives to the ethidium bromide staining procedure. MATERIALS AND METHODS: Bacterial colonization was attained in situ on bovine enamel slabs (n = 6 subjects). Five different live/dead assays were investigated (fluorescein diacetate (FDA)/propidium iodide (PI), Syto 9/PI (BacLight®), FDA/Sytox red, Calcein acetoxymethyl (AM)/Sytox red, and carboxyfluorescein diacetate (CFDA)/Sytox red). After 120 min of oral exposure, analysis was performed with an epifluorescence microscope. Validation was carried out, using the colony-forming units for quantification and the transmission electron microscopy for visualization after staining. RESULTS: The average number of bacteria amounted to 2.9 ± 0.8 × 10(4) cm(-2). Quantification with Syto 9/PI and Calcein AM/Sytox red yielded an almost equal distribution of cells (Syto 9/PI 45% viable, 55% avital; Calcein AM/Sytox red 52% viable, 48% avital). The live/dead ratio of CFDA/Sytox red and FDA/Sytox red was 3:2. An aberrant dispersal was recorded with FDA/PI (viable 34%, avital 66%). The TEM analysis indicated that all staining procedures affect the structural integrity of the bacterial cells considerably. CONCLUSION: The following live/dead assays are reliable techniques for differentiation of viable and avital adherent bacteria: BacLight, FDA/Sytox red, Calcein AM/Sytox red, and CFDA/Sytox red. These fluorescence-based techniques are applicable alternatives to toxic and instable conventional assays, such as the staining procedure based on ethidium bromide. CLINICAL RELEVANCE: Differentiation of viable and avital adherent bacteria offers the possibility for reliable evaluation of different mouth rinses, oral medication, and disinfections.

Erosion behaviour of human, bovine and equine dental hard tissues.

Dental hard tissues from different species are used in dental research, but little is known about their comparability. The aim of this study was to compare the erosive behaviour of dental hard tissues (enamel, dentin) obtained from human, bovine and equine teeth. In addition, the protective effect of the pellicle on each hard tissue under erosive conditions was determined. In situ pellicle formation was performed for 30 min on enamel and dentin samples from all species in four subjects. Calcium and phosphate release was assessed during 120 s of HCl incubation on both native and pellicle-covered enamel and dentin samples. SEM and TEM were used to examine surface changes in native enamel and dentin samples after acid incubation and the ultrastructure of the pellicle before and after erosive exposure. In general, bovine enamel and dentin showed the highest degree of erosion after acid exposure compared to human and equine samples. Erosion of human primary enamel tended to be higher than that of permanent teeth, whereas dentin showed the opposite behaviour. SEM showed that eroded equine dentin appeared more irregular than human or bovine dentin. TEM studies showed that primary enamel appeared to be most susceptible to erosion.

Enzyme activities in the oral fluids of patients suffering from bulimia: a controlled clinical trial.

Patients with bulimia nervosa are at high risk for dental erosion. However, not all bulimic patients suffer from erosion, irrespective of the severity of their eating disorder. It is often speculated that differences in the saliva are important, however, little is known about salivary parameters in bulimic patients, particularly directly after vomiting. The aim of the clinical trial was to compare different salivary parameters of subjects suffering from bulimia with those of healthy controls. Twenty-eight subjects participated (14 patients with bulimia nervosa, 7 of them with erosion; 14 matched healthy controls). Resting and stimulated saliva of all participants was analysed as well as saliva collected from bulimic patients directly and 30 min after vomiting. Parameters under investigation were flow rate, pH, buffering capacity and the enzyme activities of proteases in general, collagenase, pepsin, trypsin, amylase, peroxidase, and lysozyme. Regarding flow rate, pH and buffering capacity only small differences were found between groups; buffering capacity directly after vomiting was significantly lower in bulimic subjects with erosion than in subjects without erosion. Differences in enzymatic activities were more pronounced. Activities of proteases, collagenase and pepsin in resting and proteases in stimulated saliva were significantly higher in bulimic participants with erosion than in controls. Peroxidase activity was significantly decreased by regular vomiting. Proteolytic enzymes seem to be relevant for the initiation and progression of dental erosion directly after vomiting, maybe by both hydrolysis of demineralized dentine structures as well as modulation of the pellicle layer.

Effect of safflower oil on the protective properties of the in situ formed salivary pellicle.

AIM: The prevalence of dental erosion is still increasing. A possible preventive approach might be rinsing with edible oils to improve the protective properties of the pellicle layer. This was tested in the present in situ study using safflower oil. METHODS: Pellicle formation was carried out in situ on bovine enamel slabs fixed buccally to individual upper jaw splints (6 subjects). After 1 min of pellicle formation subjects rinsed with safflower oil for 10 min, subsequently the samples were exposed in the oral cavity for another 19 min. Enamel slabs without oral exposure and slabs exposed to the oral cavity for 30 min without any rinse served as controls. After pellicle formation in situ, slabs were incubated in HCl (pH 2; 2.3; 3) for 120 s, and kinetics of calcium and phosphate release were measured photometrically (arsenazo III, malachite green). Furthermore, the ultrastructure of the pellicles was evaluated by transmission electron microscopy (TEM). RESULTS: Pellicle alone reduced erosive calcium and phosphate release significantly at all pH values. Pellicle modification by safflower oil resulted in an enhanced calcium loss at all pH values and caused an enhanced phosphate loss at pH 2.3. TEM indicated scattered accumulation of lipid micelles and irregular vesicle-like structures attached to the oil-treated pellicle layer. Acid etching affected the ultrastructure of the pellicle irrespective of oil rinsing. CONCLUSION: The protective properties of the pellicle layer against extensive erosive attacks are limited and mainly determined by pH. The protective effects are modified and reduced by rinses with safflower oil.

Nanotechnology and its role in caries therapy.

The purpose of this review is to highlight recent nanotechnological developments for remineralization of incipient caries lesions as well as biomimetic strategies for enamel synthesis based on the application of nanotechnology. Analysis of in vitro data indicates that apatite nanoparticles might be effective in reversing lesion progression in the outer but not in the deeper part of early caries lesions. To control caries-induced demineralization, investigators have developed calcium and phosphate or fluoride ion-releasing nanofillers, enabling resin composites to release ions, if the pH decreases under in vitro conditions. Extensive in vitro investigations of apatite crystallization have been performed to mimic the hierarchical topology of natural enamel. Strategies for formation of highly organized biomineralized structures include oriented aggregation of nanocrystallites or the assembly of apatite nanoparticles mediated by organic scaffolds. Despite all these promising in vitro experiments, the effectiveness of such strategies for the control of demineralization processes as well as for caries therapy still needs validation by clinical studies.

Taste perception in children with different caries activity.

PURPOSE: The aim of the study was to investigate whether caries-active and caries-free children differ in terms of their taste perception for sweet, sour, salty and bitter. METHODS: The study group consisted of 100 children aged 5-10 years: 50 caries-free children and 50 children with at least four untreated carious lesions. Taste perception was analysed using two test systems. First, filter paper strips impregnated with four taste qualities (sweet, sour, salty, and bitter) at four defined intensities were used ("taste strips"). Then a tasting spray in suprathreshold concentration of the respective taste was applied. The children were asked to name the perceived taste. The parents answered a questionnaire about the children's dietary habits. RESULTS: The children with high caries activity scored lower on average on the taste strips than the caries-free participants. For the taste sprays, the perception of the taste quality "bitter" was significantly worse in the children with caries than in caries-free children (Mann-Whitney U test p < 0.05). CONCLUSION: The results of this study suggest that taste preferences in children are associated with increased susceptibility to tooth decay.

The lipid composition of the in situ pellicle.

OBJECTIVE: The present study aimed to systematically analyse the complete lipid profile of the in situ pellicle in comparison to saliva. For the first time, the modern sensitive methods GC-EI/MS and HPLC MS/MS were to be used for this purpose. DESIGN: Bovine enamel slabs were exposed to the oral cavity of 12 subjects by customized splints (3 min, 30 min or 120 min). Afterwards, the pellicle samples were obtained and further investigated in vitro. Additionally, corresponding unstimulated saliva samples were collected. GC-EI/MS was performed to qualitatively and quantitatively determine all fatty acids contained in the investigated samples. The individual lipid classes of phospholipids, triacylglycerols, glycolipids, cholesterol and cholesterol esters were analysed qualitatively by HPLC MS/MS. RESULTS: A characteristic fatty acid profile of the in situ pellicle was proven. Furthermore, triacylglycerols with the major fatty acids 16:0, 18:0, 18:1, 18:2, and phospholipids were detected as integral components in the pellicle. There were four groups of phospholipids: Lyso-phosphatidylcholines, phosphatidylcholines, phosphatidylethanol-amines, and phosphatidylinositols. Differences between saliva and pellicle were evident in the composition of the fatty acid- and the phospholipid profile. Glycolipids, cholesterol and cholesterol esters could neither be detected in pellicle- nor in saliva samples. CONCLUSION: The lipid profiles of the in situ pellicle and saliva were successfully characterised. Differences in the phospholipid and fatty acid composition between pellicle and saliva indicate a selective pellicle formation process. The results provide an important reference and core data for further investigation of the complex surface interactions in the oral cavity, especially concerning hydrophobic substances.

Microhardness of composite materials with different organic phases in deep class II cavities: an in vitro study.

OBJECTIVE: This study compared the microhardness of three composite resins with different organic matrices in deep class II cavities. MATERIALS AND METHOD: A total of 36 extracted molars were randomly assigned to six groups and standardized class II cavities were prepared. The cavity design comprised three steps in a mesiodistal direction with an increasing depth (2, 4, and 6 mm). Twelve cavities each were restored using Filtek Supreme (FS), Quixfil (QF), and Filtek Silorane (SI). The materials were applied in incremental layers of 2 mm and cured either with Halogen Translux Energy (HTE) (n=18) or LED Bluephase C8 (LED) (n=18). Subsequently, the specimens were cross-sectioned, and microhardness was determined in various depths and at two different distances from the matrix. RESULTS: QF yielded the highest KHN microhardness values (92.67 ± 12.77), followed by FS (65.53 ± 19.52) and SI (57.67 ± 8.33). Composites cured with LED achieved higher KHN values. All materials showed the highest microhardness values within the superficial increments and at a distance of 1000 µm from the matrix.

Characterization of the in situ pellicle ultrastructure formed under the influence of bovine milk and milk protein isolates.

OBJECTIVES: The present study aimed to investigate if bovine milk or milk protein isolates, respectively, alter the ultrastructure of thein situ pellicle and might therefore have an influence on oral health. METHODS: In situ pellicle samples were formed on bovine enamel slabs exposed in the oral cavity of three subjects for 6, 30, 60 or 120 min. After 3 min of pellicle formation, mouthrinses were performed for 3 min with (non-)homogenized UHT- or fresh milk (0.3% or 3.8% fat), 30% UHT-treated cream or different types of casein- or milk protein isolates containing preparations. The specimens were removed after the exposure times and transmission electron microscopy (TEM) was performed. Native pellicle samples served as controls. RESULTS: Topical ultrastructural pellicle modifications were detected after mouthrinses with all types of homogenized UHT- or fresh milk and after the application of a 3% native casein micelles containing experimental solution. Atypical globular protein structures, identified as casein micelles, were temporarily adsorbed onto the pellicle. They were closely associated with lipid droplets. Furthermore, the mouthrinses occasionally affected the morphology of salivary bacteria. However, no notable ultrastructural alterations remained after 120 min of pellicle formation. CONCLUSION: For the first time, bovine milk- and micellar casein-induced pellicle modifications were revealed by TEM. The adsorption of micellar casein is possibly due to its molecular interactions. CLINICAL SIGNIFICANCE: Bovine milk or micellar caseins provide some potential for the development of preventive strategies against bacterial biofilm formation or erosive processes at the tooth surface.

Fluorescence microscopic visualization and quantification of initial bacterial colonization on enamel in situ.

OBJECTIVE: The acquired salivary pellicle has been defined as proteinaceous film free of bacteria. However, due to the large numbers of microorganisms existent in the oral fluids, it is conceivable that adherent bacteria are already present in the initial pellicle. The aim of this in situ study was to visualize and to quantify these bacteria. DESIGN: Initial biofilm formation was performed on bovine enamel slabs mounted buccally on individual splints and carried in situ by six subjects for 3, 30 and 120 min, respectively. After intraoral exposure, the slabs were rinsed with saline solution and the adherent bacteria were investigated with the following fluorescence microscopic methods: staining with 4',6-diamidino-2-phenylindole (DAPI), staining of vital and nonvital bacteria with fluoresceinediacetate and ethidiumbromide (live/dead staining) and fluorescence in situ hybridization (FISH) of eubacteria and streptococci, respectively. In addition, determination of colony forming units after ultrasonically induced detachment of bacteria was performed. RESULTS: With all the methods, bacteria were detected in the initial in situ biofilm irrespective of the formation time. The numbers of bacteria revealed high intraindividual and interindividual variability and the microorganisms were distributed randomly in small aggregates. The results of the epifluorescence microscopic techniques corresponded well. The mean number of adherent bacteria detected was in the range of 10-20x10(4)cm(-2). CONCLUSION: Already after 3 min, adherent bacteria are present in the initial pellicle. For the first time, DAPI-staining as well as FISH have proven success for visualization of initial intraoral colonization of enamel specimens.

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

OBJECTIVE: The present in situ - investigation aimed to specify the impact of pure hydroxyapatite microclusters on initial bioadhesion and bacterial colonization at the tooth surface. DESIGN: Pellicle formation was carried out in situ on bovine enamel slabs (9 subjects). After 1min of pellicle formation rinses with 8ml 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 8h 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. RESULTS: Compared to the control (2.36×106±2.01×106bacteria/cm2), significantly reduced amounts of adherent bacteria were detected on specimens rinsed with chlorhexidine 0.2% (8.73×104±1.37×105bacteria/cm2) and likewise after rinses with the hydroxyapatite testsolution (2.08×105±2.85×105bacteria/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. CONCLUSION: Hydroxyapatite microclusters reduced initial bacterial adhesion to enamel in situ considerably and could therefore sensibly supplement current approaches in dental prophylaxis.

Decision criteria for replacement of fillings: a retrospective study.

The exchange of restorations goes along with the loss of healthy tooth structure. Therefore, it is important to investigate helpful decision criteria for the replacement of fillings. Five hundred forty-four filling replacements were evaluated retrospectively. Thereby, different clinical parameters were correlated with the clinical finding of caries directly after removal of the existing filling. The parameters checked for correlations were amalgam and composite, age, and size of the filling, morphology, condition of the filling, type of caries, oral hygiene, anamnesis of the respective tooth, and the decisive factor to replace the restoration. Statistical evaluation was performed by chi-squared-test (P < 0,05) and by regression analysis (Power: 80%). A percentage of 69.8% of all cavities showed softened dentin if exploring with the probe after the removal of the restoration, 7.6% were stainable with caries detector, and 22.6% of the cavities were caries free. Significant indicators for a carious lesion were high age of restoration, imperfections at the margin of the filling, a positive pain sensation in correlation with composite fillings, and multi-surface amalgam fillings. On suspicion of caries, the following decision criteria should encourage the dentist to remove a filling: High age of the filling, imperfections at the margin of the filling, especially fillings with marginal cracks, visible secondary caries, a positive pain sensation in composite filled teeth, and multi-surface amalgam fillings. Filling removals only performed due to the patient's desire for removal should be critically regarded, as most of these fillings are caries free.