Efficacy of a Mouthwash Containing CHX and CPC in SARS-CoV-2-Positive Patients: A Randomized Controlled Clinical Trial.

Soon after the outbreak of the coronavirus disease 2019 (COVID-19) pandemic, preprocedural mouthwashes were recommended for temporarily reducing intraoral viral load and infectivity of individuals potentially infected with the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) in order to protect medical personnel. Particularly, the antiseptic cetylpyridinium chloride (CPC) has shown virucidal effects against SARS-CoV-2 in vitro. Therefore, the aim of this randomized controlled clinical trial was to investigate the efficacy of a commercially available mouthwash containing CPC and chlorhexidine digluconate (CHX) at 0.05% each in SARS-CoV-2-positive patients as compared to a placebo mouthwash. Sixty-one patients who tested positive for SARS-CoV-2 with onset of symptoms within the last 72 h were included in this study. Oropharyngeal specimens were taken at baseline, whereupon patients had to gargle mouth and throat with 20 mL test or placebo (0.9% NaCl) mouthwash for 60 s. After 30 min, further oropharyngeal specimens were collected. Viral load was analyzed by quantitative reverse transcriptase polymerase chain reaction, and infectivity of oropharyngeal specimens was analyzed by virus rescue in cell culture and quantified via determination of tissue culture infectious doses 50% (TCID50). Data were analyzed nonparametrically (α = 0.05). Viral load slightly but significantly decreased upon gargling in the test group (P = 0.0435) but not in the placebo group. Viral infectivity as measured by TCID50 also significantly decreased in the test group (P = 0.0313), whereas there was no significant effect but a trend in the placebo group. Furthermore, it was found that the specimens from patients with a vaccine booster exhibited significantly lower infectivity at baseline as compared to those without vaccine booster (P = 0.0231). This study indicates that a preprocedural mouthwash containing CPC and CHX could slightly but significantly reduce the viral load and infectivity in SARS-CoV-2-positive patients. Further studies are needed to corroborate these results and investigate whether the observed reductions in viral load and infectivity could translate into clinically useful effects in reducing COVID-19 transmission (German Clinical Trials Register DRKS00027812).

Histology of human teeth: Standard and specific staining methods revisited.

OBJECTIVE: Histological techniques have long been an integral part of dental research. Especially the processing of complex tissues poses specific challenges, however, literature offers only few technical references. Objectives of this study were therefore to optimize histological staining methods and compile detailed protocols for preparation and staining of dental tissues. METHODS: Human teeth were collected and fixed with 4 % formaldehyde solution after extraction. Subsequently, teeth were decalcified in 17 % EDTA or Morse's solution over a period of 28 days. The extent of decalcification was determined by weight loss and radiography. After sectioning, histological staining methods were optimized for their use on teeth. These included hematoxylin-eosin, Masson trichrome, Masson-Goldner trichrome and May-Gruenwald-Giemsa staining. Nerve fibres were visualized by luxol fast blue staining and Bodian silver staining. In addition, specific methods like TRAP, modified Brown and Brenn as well as picrosirius red staining with light polarization or fluorescence were applied and optimized. RESULTS: Preparation of an artificial access to the pulp chamber was essential to ensure prompt penetration of the chemicals. Decalcification with Morse's solution took at least two weeks but was more efficient than 17 % ETDA, where thorough demineralization was achieved only after three weeks. The staining methods exhibited differences not only regarding their ability to display specific structures of interest, but also in terms of reproducibility. CONCLUSION: High-quality histology of teeth can only be achieved after optimal tissue preparation and accurate staining. A complementary use of staining techniques is necessary to answer specific research questions.

Penetration depth of irrigants into root dentine after sonic, ultrasonic and photoacoustic activation.

AIM: To compare penetration depths of endodontic irrigants into the dentinal tubules of extracted teeth when using several activation methods. METHODOLOGY: The root canals of 90 extracted human teeth were prepared to size 40, .06 taper. The straight and round-shaped root canals were distributed randomly into six groups, and final irrigation was performed with EDTA and sodium hypochlorite as follows: (I) manual dynamic activation, (II) Ultrasonic, (III) Sonic, (IV) PIPS (photon-induced photoacoustic streaming, (V) SWEEPS (shock-wave enhanced emission photoacoustic streaming) and (0) control without final irrigation or activation. Subsequently, methylene blue was inserted into the canals and activated according to the groups (I-V). Teeth were sectioned horizontally, imaged under a light microscope, and dye penetration depths were measured in six sections per tooth and 24 points on a virtual clock-face per section. Data were analysed statistically by nonparametric tests for whole teeth and separately for coronal, middle and apical thirds. RESULTS: Penetration of dye into the dentinal tubules was lowest for the controls. Median penetration depths amounted to 700-900 µm for groups I-V with differences in the apical thirds between group I and the other test groups. Minimum penetration depths were significantly greater for PIPS in the apical thirds (P ≤ 0.046). CONCLUSIONS: Greater penetration depths occurred in the apical thirds for ultrasonic, sonic and laser-induced activation compared to manual dynamic activation. PIPS was associated with deeper penetration of irrigants. The novel SWEEPS mode did not increase irrigant penetration.

Interactive effects of LPS and dentine matrix proteins on human dental pulp stem cells.

AIM: To investigate the combinatorial effects of lipopolysaccharide (LPS) and extracted dentine matrix proteins (eDMP) on regenerative and inflammatory responses in human dental pulp stem cells (DPSCs). METHODOLOGY: Culture media were supplemented with several concentrations of LPS, eDMP and combinations of both. Cell viability was assessed over 1 week by MTT assay; cell survival was evaluated after 24 h and 7 days by flow cytometry. The expression of mineralization-associated marker genes was determined by real-time quantitative polymerase chain reaction (RT-qPCR). To analyse the inflammatory response, secretion of interleukin 6 (IL-6) was quantified in the initial and the late phase of cell culture by enzyme-linked immunosorbent assay (ELISA). Data were treated nonparametrically and Mann-Whitney U-tests were performed to compare all experimental groups (α = 0.05). RESULTS: Whereas LPS had no impact on viability, eDMP led to a concentration-dependent decrease, which was significant after 7 days (P ≤ 0.024). A moderate decline of cell survival induced by LPS was detected after 48 h (P ≤ 0.026), whereas eDMP was able to reverse this effect. eDMP alone caused increased expression of tested marker genes, LPS had no regulatory effect. Combined eDMP and LPS induced an upregulation of collagen type I and osteocalcin, whereas expression levels of dentine matrix acidic phosphoprotein and dentine sialophosphoprotein were similar to the control. IL-6-secretion was increased by LPS over time. eDMP markedly elevated initial production of IL-6 (P ≤ 0.002), but suppressed LPS-induced cytokine production in the later phase. CONCLUSIONS: Lipopolysaccharide did not affect cell viability but interfered with odontoblast-like cell differentiation of DPSCs. Proteins from the dentine matrix may have a protective effect, attenuate the detrimental impact of LPS and thus play an important role during pulp repair.

Dentine matrix proteins: isolation and effects on human pulp cells.

AIM: To establish a simplified and efficient protocol for the isolation and concentration of matrix proteins from human dentine, and to assess the effects of extracted dentine matrix proteins (eDMP) on the behaviour of human pulp cells. METHODOLOGY: Matrix proteins were isolated from human dentine, purified, concentrated and characterized with protein and enzyme-linked immunosorbent assays (ELISA). Culture media were supplemented with eDMP in different concentrations, referred to as eDMP 1-10 000, to assess viability and proliferation of human pulp cells by DNA and MTT assays; apoptotic events were quantified by flow cytometry. Chemotactic effects of eDMP were assessed in a modified Boyden chamber assay. Expression levels of odontoblastic marker genes in pulp cells cultured with eDMPs were determined by real-time quantitative PCR, and the ability to induce mineralization was demonstrated by alizarin red staining. Nonparametric statistical analysis was performed to pairwise compare different groups at all time-points (Mann-Whitney U-test, α = 0.05). RESULTS: High concentrations of eDMP exhibited significant antiproliferative effects (P ≤ 0.023) after 5 (eDMP 1000) and 7 days (eDMP 500) without affecting cell viability. Apoptosis was barely influenced (P ≥ 0.089). eDMP exerted a concentration-dependent chemotactic stimulus on dental pulp cells with statistical significance already at low dosage (P = 0.006 at eDMP 10). Changes in gene expression indicated a differentiation into odontoblast-like cells, which was corroborated by findings of mineral nodule formation. CONCLUSIONS: A novel, effective and time-saving protocol for isolation and concentration of dentine matrix proteins is presented. As eDMP stimulates chemotaxis, differentiation and mineralization without affecting viability, endogenous dentine matrix proteins might be valuable for approaches to regenerate or engineer dental pulp.

A training model for revitalization procedures.

AIM: To develop an inexpensive simulation model for training of revitalization procedures. METHODOLOGY: A replica of an immature maxillary central incisor was equipped with a mock blood reservoir at the root apex and embedded in a plaster model. Mock blood consisted of water supplemented with red pigments and fibrinogen, whilst thrombin was inserted into the root canal to allow for clot formation. RESULTS: A true-to-life training model for revitalization procedures was established, where the induction of bleeding and formation of a blood clot can be mimicked. The model can be fastened to a patient mannequin and thus closely simulate the clinical setting. CONCLUSIONS: The newly developed model can be used for training purposes, both for dental students and for practitioners, to perform the treatment steps of a revitalization procedure.

Critical role of superoxide anions and hydroxyl radicals in HEMA-induced apoptosis.

OBJECTIVE: Resin monomers such as 2-hydroxyethyl methacrylate (HEMA) induce apoptosis because of the excess formation of reactive oxygen species (ROS). The portion of ROS including superoxide anions, hydrogen peroxide or hydroxyl radicals in monomer-induced apoptosis is unknown. Here, the effectiveness of superoxide anions or hydroxyl radicals was analyzed using tempol or sodium formate as radical scavengers. METHODS: RAW264.7 mouse macrophages were exposed to HEMA (0-6-8mM) in the presence of tempol (0-0.05-0.5-5.0mM) or sodium formate (0-1-5-10mM). The formation of ROS using DCFH2-DA or dihydrorhodamine 123 (DHR123) as fluorescent dyes and the induction of apoptosis was determined by flow cytometry after 1h or 24h exposure periods. Expression of enzymes related to ROS metabolism was detected by Western blotting. RESULTS: DCF fluorescence significantly increased after short exposure (1h) while DHR123 fluorescence was enhanced after a long exposure period (24h) in cells treated with HEMA. Although no influence was detected on the formation of ROS, tempol or sodium formate protected cells from HEMA-induced apoptosis. The number of cells in late apoptosis or necrosis induced with 6 or 8mM HEMA was reduced in the presence of tempol or low concentrations of sodium formate. HEMA-induced expression of catalase, indicating oxidative stress, decreased in the presence of tempol. SIGNIFICANCE: Superoxide anions and hydroxyl radicals contribute to HEMA-induced apoptosis. The current findings support the development of strategies based on the pharmacological inhibition of enzymes producing superoxide anions finally converted to hydroxyl radicals to compensate for potential adverse tissue reactions associated with dental composites.

Ultrasonic activation of irrigants increases growth factor release from human dentine.

OBJECTIVES: Bioactive proteins are sequestered in human dentine and play a decisive role in dental pulp regeneration and repair. They can be released and exposed on the dentine surface by acids, but also chelators, such as ethylenediaminetetraacetic acid (EDTA). The objectives of this study were (i) to evaluate whether ultrasonic activation of irrigants in the root canal will promote growth factor release from dentine and (ii) to collect bioactive proteins in a physiological solution. MATERIALS AND METHODS: Human dentine disks underwent irrigation with and without ultrasonic activation. The protocols included treatment by either a single or two consecutive steps with 10 % EDTA and phosphate-buffered saline (PBS), where each sample was treated three times. To mimic clinical conditions, selected irrigation regimens were applied to root canals of extracted human teeth after preparation. Amounts of transforming growth factor ß1 (TGF-ß1) in solution were quantified using enzyme-linked immunosorbent assays. Nonparametric statistical analysis was performed to compare different groups as well as repetitions within a group (Mann-Whitney U test, α = 0.05). Additionally, morphological changes of dentine surfaces were visualized by scanning electron microscopy (SEM). RESULTS: TGF-ß1 was not detectable after irrigation of dentine with PBS, neither with nor without ultrasonic activation. Irrigation with EDTA released TGF-ß1, and ultrasonic activation of EDTA enhanced this effect. However, preceding EDTA conditioning enabled the release of bioactive proteins into PBS solution. Similar results were observed in dentine disks and root canals. Visualization of dentine surfaces after different treatment revealed superficial erosion after ultrasonic activation irrespective of the irrigant solution, but different degrees of exposure of organic substance. CONCLUSIONS: Ultrasonic activation enhances growth factor release from human dentine. Bioactive proteins can be isolated in physiological solvents and may act as autologous supplements for regenerative endodontic treatment or pulp tissue engineering. CLINICAL RELEVANCE: Autologous growth factors from human dentine can advance treatment strategies in dental pulp tissue engineering.

EDTA conditioning of dentine promotes adhesion, migration and differentiation of dental pulp stem cells.

AIM: To evaluate the effect of dentine conditioning on migration, adhesion and differentiation of dental pulp stem cells. METHODOLOGY: Dentine discs prepared from extracted human molars were pre-treated with EDTA (10%), NaOCl (5.25%) or H2 O. Migration of dental pulp stem cells towards pre-treated dentine after 24 and 48 h was assessed in a modified Boyden chamber assay. Cell adhesion was evaluated indirectly by measuring cell viability. Expression of mineralization-associated genes (COL1A1, ALP, BSP, DSPP, RUNX2) in cells cultured on pre-treated dentine for 7 days was determined by RT-qPCR. Nonparametric statistical analysis was performed for cell migration and cell viability data to compare different groups and time-points (Mann-Whitney U-test, α = 0.05). RESULTS: Treatment of dentine with H2 O or EDTA allowed for cell attachment, which was prohibited by NaOCl with statistical significance (P = 0.000). Furthermore, EDTA conditioning induced cell migration towards dentine. The expression of mineralization-associated genes was increased in dental pulp cells cultured on dentine after EDTA conditioning compared to H2 O-pre-treated dentine discs. CONCLUSIONS: EDTA conditioning of dentine promoted the adhesion, migration and differentiation of dental pulp stem cells towards or onto dentine. A pre-treatment with EDTA as the final step of an irrigation protocol for regenerative endodontic procedures has the potential to act favourably on new tissue formation within the root canal.

Three-dimensional culture of dental pulp stem cells in direct contact to tricalcium silicate cements.

OBJECTIVES: Calcium silicate cements are biocompatible dental materials applicable in contact with vital tissue. The novel tricalcium silicate cement Biodentine™ offers properties superior to commonly used mineral trioxide aggregate (MTA). Objective of this study was to evaluate its cytocompatibility and ability to induce differentiation and mineralization in three-dimensional cultures of dental pulp stem cells after direct contact with the material. MATERIALS AND METHODS: Test materials included a new tricalcium silicate (Biodentine™, Septodont, Saint-Maur-des-Fossés, France), MTA (ProRoot® MTA, DENSPLY Tulsa Dental Specialities, Johnson City, TN, USA), glass ionomer (Ketac™ Molar Aplicap™, 3M ESPE, Seefeld, Germany), human dentin disks and polystyrene. Magnetic activated cell sorting for to the surface antigen STRO-1 was performed to gain a fraction enriched with mesenchymal stem cells. Samples were allowed to set and dental pulp stem cells in collagen carriers were placed on top. Scanning electron microscopy of tricalcium silicate cement surfaces with and without cells was conducted. Cell viability was measured for 14 days by MTT assay. Alkaline phosphatase activity was evaluated (days 3, 7, and 14) and expression of mineralization-associated genes (COL1A1, ALP, DSPP, and RUNX2) was quantified by real-time quantitative PCR. Nonparametric statistical analysis for cell viability and alkaline phosphatase data was performed to compare different materials as well as time points (Mann-Whitney U test, α = 0.05). RESULTS: Cell viability was highest on tricalcium silicate cement, followed by MTA. Viability on glass ionomer cement and dentin disks was significantly lower. Alkaline phosphatase activity was lower in cells on new tricalcium silicate cement compared to MTA, whereas expression patterns of marker genes were alike. CONCLUSIONS: Increased cell viability and similar levels of mineralization-associated gene expression in three-dimensional cell cultures on the novel tricalcium silicate cement and mineral trioxide aggregate indicate that the material is cytocompatible and bioactive. CLINICAL RELEVANCE: The tested new tricalcium silicate cement confirms its suitability as an alternative to MTA in vital pulp therapy.

Cerium chloride reduces enamel lesion initiation and progression in vitro.

AIM: Determination of the potential of cerium chloride to reduce artificial carious mineral loss and lesion depth progression. METHODS: A total of 160 enamel samples were prepared from 40 bovine lower central incisors. Crowns were sectioned into four pieces, embedded in acrylic resin, ground flat and allocated to eight groups (S1-S4 and D1-D4; n = 20). Specimens of groups D1-D4 were stored (for 7 days) in a demineralizing buffer solution to induce caries-like lesions. Afterwards, samples were treated for 30 s with one of the following solutions: placebo (S1 and D1), amine fluoride (S2 and D2), cerium chloride (S3 and D3) and a combination of fluoride and cerium chloride (S4 and D4). After another 7 (D1-D4) or 14 (S1-S4) days in demineralizing buffer solution, integrated mineral loss and lesion depth were determined by transversal microradiography and compared by Scheffé's post hoc tests. RESULTS: In groups S1-S4, the highest values for integrated mineral loss and lesion depth were observed for group S1 (placebo), the lowest values for group S4. The results in groups S2-S4 were not significantly different. In groups D1-D4, the highest values for integrated mineral loss and lesion depth were observed for group D1 (placebo), the lowest values in groups D3 and D4. In group D2, integrated mineral loss and lesion depth were significantly lower as compared to D1, but significantly higher compared to groups D3 and D4. CONCLUSION: Cerium chloride and its combination with fluoride are able to significantly reduce carious mineral loss and the progression of lesion depth.

Different protocols to produce artificial dentine carious lesions in vitro and in situ: hardness and mineral content correlation.

This study compared dentine demineralization induced by in vitro and in situ models, and correlated dentine surface hardness (SH), cross-sectional hardness (CSH) and mineral content by transverse microradiography (TMR). Bovine dentine specimens (n = 15/group) were demineralized in vitro with the following: MC gel (6% carboxymethylcellulose gel and 0.1 M lactic acid, pH 5.0, 14 days); buffer I (0.05 M acetic acid solution with calcium, phosphate and fluoride, pH 4.5, 7 days); buffer II (0.05 M acetic acid solution with calcium and phosphate, pH 5.0, 7 days), and TEMDP (0.05 M lactic acid with calcium, phosphate and tetraethyl methyl diphosphonate, pH 5.0, 7 days). In an in situ study, 11 volunteers wore palatal appliances containing 2 bovine dentine specimens, protected with a plastic mesh to allow biofilm development. The volunteers dripped a 20% sucrose solution on each specimen 4 times a day for 14 days. In vitro and in situ lesions were analyzed using TMR and statistically compared by ANOVA. TMR and CSH/SH were submitted to regression and correlation analysis (p < 0.05). The in situ model produced a deep lesion with a high R value, but with a thin surface layer. Regarding the in vitro models, MC gel produced only a shallow lesion, while buffers I and II as well as TEMDP induced a pronounced subsurface lesion with deep demineralization. The relationship between CSH and TMR was weak and not linear. The artificial dentine carious lesions induced by the different models differed significantly, which in turn might influence further de- and remineralization processes. Hardness analysis should not be interpreted with respect to dentine mineral loss.

A survey on education in cariology for undergraduate dental students in Europe.

The aim of the survey was to collect relevant information about education in cariology for dental undergraduate students in Europe. The ORCA/ADEE cariology curriculum group prepared a questionnaire that was mailed in 2009 to 179 European dental schools. One hundred and twenty-three dental schools (72%) from 32 countries completed and returned the questionnaires. In most of these schools, education in cariology is delivered by at least two different units. The units mainly involved are Paediatric Dentistry, Conservative Dentistry, Restorative Dentistry or Operative Dentistry. Theoretical education in cariology is delivered by practically all responding dental schools, and in 96% of these schools, it starts within the first 3 years. Pre-clinical exercises are offered by 98% of the schools starting mainly in the third or fourth year. In 97% of the schools, clinical exercises are carried out, and this occurs mainly in the fourth and fifth year. In nearly all dental schools (88%), education in cariology comprises not only caries but also dental erosion and non-erosive wear. The vast majority of the responding dental schools (89%) supported the idea of developing a European Core Curriculum in Cariology.

European Core Curriculum in Cariology for undergraduate dental students.

As dental caries prevalence is still high in many populations and groups of both children and adults worldwide, and as caries continues to be responsible for significant health, social and economic impacts, there is an urgent need for dental students to receive a systematic education in cariology based upon current best evidence. Although European curriculum guidelines for undergraduate students have been prepared in other dental fields over the last decade, none exist for cariology. Thus the European Organisation for Caries Research (ORCA) formed a task force to work with the Association of Dental Education in Europe (ADEE) on a European Core Curriculum in Cariology. In 2010, a workshop to develop such a curriculum was organised in Berlin, Germany, with 75 participants from 24 European and 3 North/South American countries. The Curriculum was debated by five pre-identified working groups: I The Knowledge Base; II Risk Assessment, Diagnosis and Synthesis; III Decision-Making and Preventive Non-surgical Therapy; IV Decision-making and Surgical Therapy; and V Evidence-based Cariology in Clinical and Public Health Practice and then finalised jointly by the group chairs. According to this Curriculum, on graduation, a dentist must be competent at applying knowledge and understanding of the biological, medical, basic and applied clinical sciences in order to recognise caries and make decisions about its prevention and management in individuals and populations. This document, which presents several major and numerous supporting competences, does not confine itself to dental caries alone, but refers also to dental erosion/non-erosive wear and other dental hard tissue disorders.

Decision-making and treatment with respect to surgical intervention in the context of a European Core Curriculum in Cariology.

This paper is part of a series of papers towards a European Core Curriculum in Cariology for undergraduate dental students. The European Core Curriculum in Cariology is the outcome of a process starting in 2006 and culminating in a joint workshop of the European Organisation for Caries Research (ORCA) together with the Association for Dental Education in Europe (ADEE) which was held in Berlin from 27 to 30 June 2010. The present paper covers decision-making and treatment with respect to surgical intervention. In particular, it will provide some background information on this part of the European Core Curriculum. Undergraduate dental education should enable the student to become a competent, skilful and caring dentist who is able, upon graduation, to take professional responsibility for diagnosis, as well as effective safe, and long-lasting care in the best interests of the patient. With respect to decision-making around surgical intervention for dental caries, several factors have to be considered. These include, patient needs, preventive strategies, tooth preservation, caries management success and failure rates, as well as short-term and long-term treatment costs. With respect to surgical intervention, manual skills at a high level are required. This is of crucial importance for the graduating dentist allowed to practise dentistry in many EU countries.

European core curriculum in cariology for undergraduate dental students.

As dental caries prevalence is still high in many populations and groups of both children and adults worldwide, and as caries continues to be responsible for significant health, social and economic impacts, there is an urgent need for dental students to receive a systematic education in cariology based upon current best evidence. Although European curriculum guidelines for undergraduate students have been prepared in other dental fields over the last decade, none exist for cariology. Thus the European Organisation for Caries Research (ORCA) formed a task force to work with the Association of Dental Education in Europe (ADEE) on a European Core Curriculum in Cariology. In 2010, a workshop to develop such a curriculum was organised in Berlin, Germany, with 75 participants from 24 European and 3 North-South American countries. The Curriculum was debated by five pre-identified working groups: I The Knowledge Base; II Risk Assessment, Diagnosis and Synthesis; III Decision-making and Preventive Non-surgical Therapy; IV Decision-making and Surgical Therapy; and V Evidence-based Cariology in Clinical and Public Health Practice and then finalised jointly by the group chairs. According to this Curriculum, on graduation, a dentist must be competent at applying knowledge and understanding of the biological, medical, basic and applied clinical sciences in order to recognise caries and make decisions about its prevention and management in individuals and populations. This document, which presents several major and numerous supporting competences, does not confine itself to dental caries alone, but refers also to dental erosion/non-erosive wear and other dental hard tissue disorders.

Effects of various fluoride solutions on enamel erosion in vitro.

The objective of this in vitro study was to investigate the effect of different fluoride solutions on enamel erosion. Human enamel specimens were pretreated with 1 of 10 different fluoride solutions (n = 20): TiF(4), NaF, AmF, ZnF(2), or SnF(2), each at native pH (pH range: 1.2-7.8) or buffered pH (pH = 4). The control group samples received no fluoride pretreatment. All samples were then eroded by citric acid (pH 2.6) for 6 x 1 min daily over 5 days. Between the erosive cycles, the samples were stored in artificial saliva. Erosion effects were investigated by surface profilometry (n = 10), scanning electron microscopy (n = 4), and energy-dispersive X-ray spectroscopy (n = 6) after fluoride pretreatment and after erosion. To test the effects of pH only, additional experiments were carried out with fluoride-free solutions at similar pH to that of fluoride solutions. In general, AmF solution was more effective in protecting enamel erosion compared to all other fluoride agents. However, the application of native TiF(4), native and buffered SnF(2), and native and buffered AmF solutions also resulted in significantly less enamel loss compared to the control group. A Ti-rich coating was formed after application of native TiF(4), but partially dissolved due to erosive attack. Samples pretreated with SnF(2) showed a significant increase in surface tin content. Surface fluoride concentration was significantly increased by native TiF(4), native and buffered AmF, buffered ZnF(2), and buffered NaF application. Under the current experimental setting, the fluoride agents at lower pH had better protective potential. Highly concentrated TiF(4), AmF, and SnF(2) solution was effective in inhibiting erosion of enamel.

Comparison of cross-sectional hardness and transverse microradiography of artificial carious enamel lesions induced by different demineralising solutions and gels.

The aims of this study were: (1) to correlate surface (SH) and cross-sectional hardness (CSH) with microradiographic parameters of artificial enamel lesions; (2) to compare lesions prepared by different protocols. Fifty bovine enamel specimens were allocated by stratified randomisation according to their initial SH values to five groups and lesions produced by different methods: MC gel (methylcellulose gel/lactic acid, pH 4.6, 14 days); PA gel (polyacrylic acid/lactic acid/hydroxyapatite, pH 4.8, 16 h); MHDP (undersaturated lactate buffer/methyl diphosphonate, pH 5.0, 6 days); buffer (undersaturated acetate buffer/fluoride, pH 5.0, 16 h), and pH cycling (7 days). SH of the lesions (SH(1)) was measured. The specimens were longitudinally sectioned and transverse microradiography (TMR) and CSH measured at 10- to 220-microm depth from the surface. Overall, there was a medium correlation but non-linear and variable relationship between mineral content and radicalCSH. radicalSH(1) was weakly to moderately correlated with surface layer properties, weakly correlated with lesion depth but uncorrelated with integrated mineral loss. MHDP lesions showed the highest subsurface mineral loss, followed by pH cycling, buffer, PA gel and MC gel lesions. The conclusions were: (1) CSH, as an alternative to TMR, does not estimate mineral content very accurately, but gives information about mechanical properties of lesions; (2) SH should not be used to analyse lesions; (3) artificial caries lesions produced by the protocols differ, especially considering the method of analysis.

Relationship between nanohardness and mineral content of artificial carious enamel lesions.

The aim of this study was to compare cross-sectional nanohardness, measured using an ultra-microindentation system, with mineral content, from transversal microradiography, of artificial enamel caries lesions. Sections (85 +/- 10 microm) from 16 bovine enamel samples with artificial caries were prepared. The mineral content and cross-sectional nanohardness at known depths from the surface were compared. Both methods showed lesion profiles with a surface layer. The determination of nanohardness seems limited to lesions with a mineral content >45 vol%. There was a moderate linear relationship between mineral content and the square root of nanohardness (R2 = 0.81). It was concluded that the conversion of cross-sectional hardness into mineral content remains questionable and cannot be recommended.