Erosion protective properties of the enamel pellicle in-situ.

OBJECTIVES: Previous studies on short- and long-term pellicles showed that the enamel pellicle provides partial protection against erosion. The aim of the present study was to investigate the protective properties of clinically relevant pellicles formed within 2 to 24 h. The hypothesis was that factors such as pellicle formation time, intraoral location, and acidic challenge severity would not influence the erosion-protective properties of the pellicle. METHODS: Six subjects participated in the study. Bovine enamel specimens were prepared and intraorally exposed at buccal or palatal sites for 2, 6, 12, and 24 h to allow pellicle formation, followed by erosion using 0.1 % or 1 % citric acid. Calcium release and surface microhardness were measured, and specimens were analysed using scanning and transmission electron microscopy. Quantitative data were statistically analysed with three-way ANOVA and Tuckey's multiple comparison test (p = 0.05). RESULTS: Pellicle formation time and intraoral location did not significantly influence the erosion-protective properties of the pellicle, while citric acid concentration significantly affected enamel erosion. The pellicle thickness increased with longer formation times and on buccal sites, but decreased or was entirely removed following treatment with 0.1 % or 1 % citric acid, respectively. The enamel surface exhibited a characteristic erosion pattern. CONCLUSIONS: This study underscores the importance of investigating pellicle properties within the critical 2- to 24-h timeframe and highlights the significance of pellicle thickness in acid resistance. CLINICAL SIGNIFICANCE: These findings provide valuable insights into the factors influencing the protective properties of enamel pellicles and could guide preventive measures in dental practice.

Effect of Polyphenols on the Ultrastructure of the Dentin Pellicle and Subsequent Erosion.

INTRODUCTION: Erosive tooth wear is a highly prevalent dental condition that is modified by the ever-present salivary pellicle. The aim of the present in situ study was to investigate the effect of polyphenols on the ultrastructure of the pellicle formed on dentin in situ and a subsequent erosive challenge. METHODS: The pellicle was formed on bovine dentin specimens for 3 min or 2 h in 3 subjects. After subjects rinsed with sterile water (negative control), 1% tannic acid, 1% hop extract, or tin/fluoride solution containing 800 ppm tin and 500 ppm fluoride (positive control), specimens were removed from the oral cavity. The erosive challenge was performed on half of the specimens with 1% citric acid, and all specimens were analyzed by transmission electron microscopy. Incorporation of tannic acid in the pellicle was investigated by fluorescence spectroscopy. RESULTS: Compared to the negative control, ultrastructural analyses reveal a thicker and electron-denser pellicle after application of polyphenols, in which, according to spectroscopy, tannic acid is also incorporated. Application of citric acid resulted in demineralization of dentin, but to a lesser degree when the pellicle was pretreated with a tin/fluoride solution. The pellicle was more acid-resistant than the negative control when modified with polyphenols or tin/fluoride solution. CONCLUSION: Polyphenols can have a substantial impact on the ultrastructure and acid resistance of the dentin pellicle, while the tin/fluoride solution showed explicit protection against erosive demineralization.

Prevention of Dental Biofilm Formation with Polyphenols: A Systematic Review.

Polyphenols are plant secondary products with health-promoting properties against various degenerative or infectious diseases, and thus may help in the prevention of oral diseases. The aim of the present systematic review was to investigate polyphenols as a possible adjuvant in inhibiting dental biofilm formation, which is an important precondition for the most prevalent oral disease - caries and periodontitis. A literature search was conducted using the databases PubMed, CENTRAL and Scopus. Only studies with oral healthy participants and plaque level as outcome were included. Data search and extraction was conducted by two authors independently. Of the 211 initially identified studies, only six met all inclusion criteria. Meta-analysis was performed with five studies using the random effect model. Treatment with polyphenols reduced the plaque level in comparison to a negative control, but not significantly. Strong evidence of heterogeneity was observed. The diversity and complexity of polyphenols and their preparation need to be considered. There is no clear evidence that clinical use of polyphenols can prevent dental biofilm formation. Additional research with more and larger randomized controlled trials are required.

Ultrastructure of the Dentin Pellicle and the Impact of Erosion.

While the ultrastructure of the enamel pellicle and its erosion protective properties are well studied, the dentin pellicle is still neglected in dental research. Therefore, the ultrastructure and erosion protective properties of a pellicle formed on bovine dentin specimens were investigated in the present study. The dentin pellicle was formed in situ for 3, 30, 120, and 360 min at buccal or palatal oral sites of 3 subjects and analyzed by transmission electron microscopy. In order to clarify the impact of an erosive challenge to the ultrastructure of the pellicle and the underlying dentin, specimens were exposed to the oral cavity and eroded in vivo with 0.1% or 1% citric acid either immediately or after 30 min of pellicle formation. Specimens that were eroded without exposure to the oral cavity served as control. In another trial, specimens with a 30-min pellicle were exposed to the oral cavity for a further 60 min after the erosive challenge to investigate the effect of saliva on the impaired pellicle and dentin. Transmission electron micrographs reveal a globular and granular structured pellicle layer, which was thicker when the pellicle was formed buccally or with longer formation times. Erosion with citric acid reduced the thickness of the pellicle and interrupted its continuity. The dentin was also affected by erosion, which was represented by a lower electron density and formation of demineralized lacunae. These were infiltrated by a granular structured material when specimens were exposed to the oral cavity. After further intraoral exposure, the infiltration was more pronounced, indicating a significant impact of saliva on the demineralized dentin. A reformation of the dentin pellicle on the other hand did not occur. In conclusion, the dentin pellicle is neither acid-resistant nor able to effectively protect dentin from erosion.

Influence of periodic polyphenol treatment on the anti-erosive potential of the acquired enamel pellicle-A qualitative exploratory study.

OBJECTIVE: The aim of the present study was to investigate the impact of periodic polyphenol treatment on the ultrastructure and anti-erosive potential of an in-situ formed pellicle. METHODS: Subjects wore intraoral appliances with buccally and palatally fixed bovine enamel specimens. During 6 h of intraoral pellicle formation, 100 ml black tea or tannic acid was applied ex-vivo every 25 min for 5 min. Untreated pellicles served as control. After the trial, specimens were immersed in 0.1% or 1% citric acid for 60 s and analysed for calcium release with atomic adsorption spectrometry and ultrastructure with transmission electron microscopy. RESULTS: Specimens covered by pellicles treated with black tea or tannic acid released less calcium than untreated pellicles. Ultrastructural analyses reveal an increase in pellicle's thickness and density after treatment with polyphenols. CONCLUSIONS: Periodic polyphenol treatment of the pellicle modify its ultrastructure and increase its anti-erosive potential. CLINICAL SIGNIFICANCE: Consumption of polyphenolic beverages can enhance the anti-erosive potential of the enamel pellicle.

Influence of a hydroxyapatite suspension on 48-h dental biofilm formation in-situ.

OBJECTIVE: The aim of the present in-situ study was to investigate anti-adherent properties of mouthrinses containing hydroxyapatite (HAP) nanoparticles on oral biofilm formation. DESIGN: Biofilm was formed for 48 h on bovine enamel or dentine specimens that were fixed to maxillary splints and worn intraoral by six volunteers. During biofilm formation, rinsing was performed with sterile water, HAP (5%) or chlorhexidine (0.2%) according to two different rinsing protocols in order to assess substantivity. Scanning electron microscopy was used to investigate biofilm coverage of specimens, biofilm thickness and morphology. In addition, saliva samples were collected and analysed with transmission electron microscopy. RESULTS: Rinsing with sterile water or HAP resulted in 2.1 or 2.3 µm thick biofilms, respectively, covering more than half of specimen' surfaces. Despite single deposits of nanoparticles in saliva and biofilm, HAP did not inhibit biofilm formation. Chlorhexidine on the other hand significantly reduced biofilm thickness and coverage. CONCLUSIONS: Mouthrinses containing HAP nanoparticles showed no anti-adherent effects during 48 h of biofilm formation in-situ.

Influence of periodic milk or cream treatment on the anti-erosive potential of the acquired enamel pellicle.

OBJECTIVE: The purpose of the present study was to assess the anti-erosive potential of the acquired enamel pellicle formed in situ under the influence of periodic milk or cream treatment. METHODS: The pellicle was formed on bovine enamel specimens in the oral cavity at buccal and palatal sites of upper molars in 6 subjects, using removable acrylic splints. During 6-h of intraoral exposure, splints were removed from the oral cavity every 25 min, treated with milk or cream for 5 min, and subsequently re-inserted into the oral cavity. After 6 h, pellicle covered specimens were immersed in citric acid (0.1 or 1.0 %) for 1 min, and processed for measurement of surface microhardness, determination of calcium release by atomic absorption spectroscopy, scanning and transmission electron microscopy. Statistical analysis was performed with SAS. RESULTS: Statistical analysis did not indicate major differences between erosive surface alterations on enamel specimens covered by pellicles treated with cream or milk, and those covered by control pellicles. In addition, TEM analysis did not reveal any differences concerning the ultrastructure of the different pellicle treatments during acid exposure. All pellicles were dissolved in part after exposure to 0.1 % citric acid and were nearly completely removed after treatment with 1.0% citric acid. CONCLUSIONS: It is concluded that periodic treatment with milk or cream during pellicle formation in situ does not improve the protective potential of the acquired enamel pellicle against erosion. CLINICAL SIGNIFICANCE: Modification of the pellicle by consumption of milk or cream prior to an acidic challenge cannot sufficiently protect enamel from erosion.

The dentin pellicle - A neglected topic in dental research.

OBJECTIVE: All soft and solid surfaces exposed to the oral cavity are covered by an acquired pellicle. While the pellicle adsorbed on enamel is well researched, only limited data are available on the dentin pellicle. The purpose of the present review is to summarize studies considering the composition, structure and properties of the dentin pellicle and compare them with the current state of research on enamel pellicle. METHODS: The literature search was conducted using Medline database and Google Scholar, including checking reference lists of journal articles by handsearching. Thereby, 19 studies were included in the present review. RESULTS AND CONCLUSION: The dentin pellicle has a similar ultrastructure to the enamel pellicle, which is up to 1 µm thick depending on pellicle formation time and localization in the oral cavity. In contrast, due to the lack of studies on the dentin pellicle regarding its composition and properties, a comparison to the enamel pellicle is difficult. So far, only one study showed anti-abrasive properties and data on anti-erosive properties were controversial. Despite becoming more and more clinically relevant due to the increasing frequency of dentin exposure, the dentin pellicle is largely unexplored. For further investigations it is not only necessary to standardize dentin specimens, but also to assess fundamental research on dentin itself, as its complex morphology and composition may have a crucial influence on pellicle formation. Furthermore, a more detailed knowledge of the dentin pellicle may also reveal target sites for modification in favor of its protective properties.

Evaluation of Anti-Biofilm Activity of Mouthrinses Containing Tannic Acid or Chitosan on Dentin In Situ.

In contrast to enamel, dentin surfaces have been rarely used as substrates for studies evaluating the effects of experimental rinsing solutions on oral biofilm formation. The aim of the present in situ study was to investigate the effects of tannic acid and chitosan on 48-h biofilm formation on dentin surfaces. Biofilm was formed intraorally on dentin specimens, while six subjects rinsed with experimental solutions containing tannic acid, chitosan and water as negative or chlorhexidine as positive control. After 48 h of biofilm formation, specimens were evaluated for biofilm coverage and for viability of bacteria by fluorescence and scanning electron microscopy. In addition, saliva samples were collected after rinsing and analyzed by fluorescence (five subjects) and transmission electron microscopy (two subjects) in order to investigate the antibacterial effect on bacteria in a planktonic state and to visualize effects of the rinsing agents on salivary proteins. After rinsing with water, dentin specimens were covered by a multiple-layered biofilm with predominantly vital bacteria. In contrast, chlorhexidine led to dentin surfaces covered only by few and avital bacteria. By rinsing with tannic acid both strong anti-adherent and antibacterial effects were observed, but the effects declined in a time-dependent manner. Transmission electron micrographs of salivary samples indicated that aggregation of proteins and bacteria might explain the antiadhesion effects of tannic acid. Chitosan showed antibacterial effects on bacteria in saliva, while biofilm viability was only slightly reduced and no effects on bacterial adherence on dentin were observed, despite proteins being aggregated in saliva after rinsing with chitosan. Tannic acid is a promising anti-biofilm agent even on dentin surfaces, while rinsing with chitosan could not sufficiently prevent biofilm formation on dentin.

Effects of Experimental Agents Containing Tannic Acid or Chitosan on the Bacterial Biofilm Formation in Situ.

Chitosan and tannic acid are known for their antibacterial properties. In the present in-situ study, their antibacterial and anti-adherent effects on biofilm formation on enamel were investigated. Six subjects carried upper jaw splints with bovine enamel specimens, allowing in-situ biofilm formation. During the two-day trial, subjects rinsed with experimental solutions that contained either chitosan, tannic acid (pH = 2.5), tannic acid (pH = 7) or hydrochloric acid. Water served as the negative and chlorhexidine as the positive control. Rinsing occurred four or five times following two different rinsing protocols to investigate both the immediate and long-lasting effects. After 48 h of intraoral exposure, the dental plaque was stained with LIVE/DEAD® BacLight, and fluorescence micrographs were evaluated by using the software ImageJ. The results were verified by scanning electron microscopy. Rinsing with chitosan resulted in little immediate antibacterial and anti-adherent effects but failed to show any long-lasting effect, while rinsing with tannic acid resulted in strong immediate and long-lasting effects. Except for a slightly lower antibacterial effect, the neutral solution of tannic acid was as good as the acidic solution. Hydrochloric acid showed neither an antibacterial nor an anti-adherent effect on dental biofilm formation. Experimental solutions containing tannic acid are promising anti-biofilm agents, irrespective of the pH values of the solutions. Chitosan, on the other hand, was not able to prevent biofilm formation.