Developments in dental plaque pH modelling.

OBJECTIVES: The objectives of this review were to give a comprehensive account of the methods used to determine dental plaque pH over the last 50 years, to review how these methods have been used in dental cariology research and to give an update as to how dental plaque pH studies might be developed in the future. DATA: Published, referred papers and abstracts of conference proceedings in the literature were reviewed. SOURCES: A comprehensive search of the electronic databases PubMed and Medline, was undertaken. In addition, a hand search of the Index Denticus was done to identify relevant citations before 1966. STUDY SELECTION: Relevant published literature in peer-reviewed publications was reviewed. No additional inclusion criteria were applied. CONCLUSIONS: This comprehensive review gives an account of the background to, history of, relative merits and demerits of, applications of and future of dental plaque pH technologies.

Quantification of dental plaque in the research environment.

OBJECTIVES: To review the established and novel methods of plaque quantification employed in dental research, including a discussion of their merits and to present a new method of planimetrically measuring plaque using light induced fluorescence. METHOD: Quantitative light-fluorescence (QLF) images were acquired from the buccal surfaces of an individual who had refrained from oral hygiene both with and without traditional plaque disclosure. Digital photographs were also taken. Images were analysed using a novel method and a percentage plaque index produced. RESULTS: Traditional plaque indices are problematic due to their integral nature and their failure to detect small, but potentially clinically relevant changes in plaque area. The use of a fluorescent technique demonstrated good reliability although there was no correlation between red fluorescent plaque and total disclosed plaque suggesting that the auto-fluorescing plaque is not a good measure of total plaque volume. CONCLUSIONS: The use of planimetric techniques can increase the power of plaque studies, potentially reducing the number of subjects and time required to separate therapies or products. Fluorescent methods of quantification have potential as they enable clear separation of the plaque covered and non-covered tooth surfaces.

The use of in situ models and QLF for the study of coronal caries.

OBJECTIVES: The purpose of the paper is to review aspects of the systems available to model the caries process in enamel. METHODS: The in situ model developed in Liverpool, and the new method of quantifying mineral loss, Quantitative Light-induced Fluorescence (QLF), are described. QLF is a powerful new diagnostic tool which can be used to measure demineralisation and remineralisation in tooth surfaces in vivo; studies to optimise, validate and use QLF in different clinical situations are described. RESULTS: Examples of the use of in situ models show that they are particularly valuable for monitoring de and remineralisation of artificial lesions in relation to product testing as alternatives to clinical trials, and present significant advances over in vitro methods. Quantification of mineral loss by Transverse Microradiography (TMR) as in the traditional Liverpool model has produced much valuable information, but the destructive nature of the method limits experimental design, and removes the system from the clinical situation. As a possible alternative, QLF has been validated and optimised. Longitudinal measures can be made on the same surface, and examples of its use are for monitoring recurrent caries and demineralisation around orthodontic brackets. CONCLUSIONS: While current in situ models provide a major advance over earlier caries models, measurement of de and remineralisation by destructive methods such as transverse microradiography limits the design of experimental investigations. QLF offers significant time saving, reduces the cost of clinical studies, and because the measurements can be carried out longitudinally in vivo, can remove the need for intra-oral appliances carrying experimental tissues.

The effect of bleaching on enamel susceptibility to acid erosion and demineralisation.

INTRODUCTION: The purpose of this in vitro study was to determine if enamel that had been bleached by carbamide (urea) peroxide gel (CPG) was at increased risk of either acid erosion or demineralisation (early caries) than un-bleached enamel. METHODS: Human incisors were employed. The samples were randomly assigned to one of 4 groups; a) 10% CPG, b) 16% CPG, c) 22% CPG and d) 10% CPG with xylitol, fluoride and potassium. Each specimen was moistened with saliva and the appropriate formulation placed for 2 hours for a total of 40 hours of exposure. In order to ensure that bleaching had taken place, tooth shades were monitored using the Shade-Eye device. Following the bleaching process, one half of the specimen was subjected to an erosive challenge, the other to a demineralisation system with one half of each sub-sample retained as a non-bleached control. Samples were assessed longitudinally with quantitative light-induced fluorescence (QLF) and at the conclusion of the study with transverse micro-radiography (TMR). RESULTS: Erosion was detected in all samples (DeltaQ 126+/-23.4), in both bleached and non-bleached areas. There was no statistical difference between the bleached and non-bleached areas either within the treatment groups or between them. Caries-like lesions were detected on all samples; TMR revealed sub-surface lesions on all teeth and QLF data supported this (DeltaQ 89+/-18.9). Following statistical analysis there were no differences detected between the bleached and non-bleached areas, nor between the different concentrations of the bleaching solution. CONCLUSION: These results suggest that tooth bleaching with carbamide (urea) peroxide (using commercially available concentrations) does not increase the susceptibility of enamel to acid erosion or caries.

Reduction in enamel dissolution by liquorice and glycyrrhizinic acid.

Liquorice extracts and confections reduced enamel dissolution in acidic buffers and saliva/glucose incubations by a direct effect on solubility, and by inhibiting the fall in pH on incubation. These actions may be attributed to the solubility-reducing, glycolysis-inhibiting, and buffering properties of glycyrrhizinic acid, a constituent of liquorice.

The relationship between plaque pH, plaque acid anion profiles, and oral carbohydrate retention after ingestion of several 'reference foods' by human subjects.

The primary aim of this study was to rank several reference foods (apple drink, caramel, chocolate, cookie, skimmed milk powder, snack cracker, and wheat flake) according to their plaque pH response as monitored in a panel of 12 volunteers by the plaque-sampling method for comparison with data previously reported with other methods used to assess cariogenicity potential. Secondary experiments (using subsets of the panel of subjects) were undertaken in an attempt to elucidate some of the reasons for the observed plaque pH changes. Oral carbohydrate retention was measured at a single time period after food use as total anthrone-positive carbohydrate material, and as specific acidogenic sugars by gas-liquid chromatography after gel-exclusion chromatography. The concentrations of acid anions in the plaque fluid after food consumption were measured by isotachophoresis eight min after food use. According to the plaque pH response, apple-flavored fruit drink and chocolate were the most acidogenic foods and skimmed milk powder the least acidogenic. There were significant correlations (p less than 0.05) between the plaque pH data and lactate-plus-acetate concentrations in plaque fluid, but the correlations between the pH data and any of the carbohydrate retention parameters were not significant.

Intra-oral models for studying de- and remineralization in man: methodology and measurement.

A wide range of different methodologies and measurement techniques has been employed in laboratories around the world for the study of de- and remineralization of enamel and dentin in intra-oral systems. These different approaches are summarized and discussed in relation to the nature of the different research questions to be studied by means of the intra-oral model.

The effect of daily gum-chewing on salivary flow rates in man.

Following reports of increased salivary gland size and increased function, induced by increased mastication in animals, the effects of long-term, frequent gum-chewing on resting and stimulated flow rates were studied in human volunteers in separate experiments in Newcastle upon Tyne and in Toronto. In both experiments, unstimulated and stimulated saliva flow rates were measured in student volunteers at intervals of one or two weeks over a baseline period. Approximately half of the subjects were then given sugarless gum to be chewed (four pieces per day) over the experimental period; controls refrained from vigorous mastication. During (and, in Newcastle, after) the experimental period, salivary flow rates were measured at intervals, as before. In Newcastle unstimulated, but not stimulated, flow rates increased in the gum-chewing group and were still elevated (compared with controls) eight weeks following the experiment. In Toronto, the mean results showed no effect of gum-chewing, but the seven gum-chewers among the 11 subjects with low baseline flow rates (less than 0.3 mL/min) showed a 43% rise in unstimulated flow rate (p approximately 0.05). The results suggest that increased mastication, in the form of gum-chewing, can increase unstimulated flow rates, especially in those with low salivary function. In addition to short-term beneficial effects of sugarless gum, these long-term effects indicate the possibility of a beneficial effect in caries prevention.

Plaque fluid as a bacterial milieu.

Studies of the extracellular, free concentrations of substrates, growth factors, inhibitors, and end-products of metabolism to which the intact plaque microflora is exposed in situ can assist in the understanding of factors controlling plaque pathogenicity. Information is becoming increasingly available from analysis of fluid separated by centrifugation of plaques collected at various intervals after an intra-oral pulse of dietary or experimental substrate, or different procedures or treatments having cariostatic potential. Such analytical results give more information than those obtained by analysis of aqueous or other extracts, because they yield values of substrate concentration representing those occurring at the bacterial cell surface. The largest body of information concerns extracellular levels of acid end-products of sugar catabolism in relation to food quality or sequence, and of amino acids and other products of nitrogen metabolism, in relation to studies of the detailed metabolic events of the Stephan curve, and of the demineralizing effect of the plaque environment. Areas where little information is available and which merit further study include plaque clearance of salivary and other components with anti-caries activity (e.g., antibodies, enzymes, fluorides, cations, other antimicrobials, etc.), and substrate concentrations to determine gradients for diffusion into and out of plaque.

The effect of altering the position of a sugary food in a meal upon plaque pH in human subjects.

Plaque pH in six adult subjects was monitored while they consumed a meal containing three foods. The marked fall in plaque pH caused by drinking sugared coffee was considerably reduced by eating the two non-sugary foods (egg and crisp-bread) before, during, or after the sugared coffee. This favorable action was negated by a pause between the sugary and non-sugary courses.

Thickness of acquired salivary pellicle as a determinant of the sites of dental erosion.

Dental erosion shows a typical distribution pattern within the dental arches. Tooth protection from erosion by salivary pellicle has been shown in vitro, but the hypothesis that pellicle may differ quantitatively at sites of erosion has not been investigated. This study aimed to determine the thickness of acquired salivary pellicle within the dental arches, investigate the possible relationship of this thickness to the distribution and severity of erosion within the arches, and confirm the protective effect of pellicle against dental erosion. Eight enamel blocks were produced from each of 5 bovine incisors assigned to five volunteers. Each block was further cut into 2 slabs, producing control and experimental slabs. Pellicle developed on experimental slabs located on 8 intra-oral sites after 1 hr of exposure was stained by "sheep anti-human IgGAM-FITC". Slabs were then visualized, and pellicle thickness measured, by confocal laser scanning microscopy. Eroded enamel lesions were produced in experimental and control slabs by means of pure orange juice. The degree of erosion was quantified by transverse microradiography. Pellicle thickness varied significantly within the dental arches and among individuals. An inverse relationship (r = -0.96, p<0.001) was observed between the degree of erosion and pellicle thickness. Significant differences in erosion were observed between slabs with and those without pellicle. This study has shown that the thickness of acquired salivary pellicle varies within the dental arches, which may be responsible for the site-specificity of dental erosion, and that pellicle does protect the teeth from erosion.

Remineralization of artificial caries-like lesions in human enamel in situ by chewing sorbitol gum.

The objective of the study was to determine quantitatively the effect on the potential for in situ remineralization of artificial caries-like lesions in human enamel when sugar-free gum containing mainly sorbitol as sweetener was chewed after meals and snacks. Artificial white-spot lesions were created in extracted human premolars and divided into three parts. One part was used as reference and the other two worn consecutively for two 21-day periods by 10 volunteers in a cast silver band cemented on lower molar teeth and covered with gauze to promote plaque formation. During the experimental periods, the subjects used fluoridated toothpaste twice daily, and consumed three meals (breakfast, lunch, and dinner) and two snacks (selected from chocolate bar, raisins, chocolate wafer, and iced cupcake). Sorbitol gum was chewed for 20 min immediately after each meal or snack during one of the experimental periods. The three parts of the enamel lesions were then sectioned (congruent to 80 microns) and examined together by means of quantitative microradiography and by polarized light microscopy. All estimates of mineral content indicated that significant remineralization occurred and was approximately doubled with gum-chewing. It is suggested that sorbitol gum stimulates salivation, which is responsible for the significantly enhanced remineralization, thus contributing to a therapeutic, caries-preventive effect. Because the gum was chewed immediately after meals and snacks, inhibition of demineralization may also have occurred.

Nosocomial infection with hepatitis A.

Nosocomial spread of hepatitis A is very uncommon but may occur under unusual circumstances, as shown by the incident described here and by the few other published reports. In this incident it is concluded that the patient, who was the index case, was excreting hepatitis A virus in the faeces 16 days before jaundice developed and 17 days before alanine aminotransferase (ALT) values reached a peak.

Effects of lactate dehydrogenase and nicotinamide adenine dinucleotide on human dental plaque pH and acid anion concentrations.

The effect of sucrose rinses supplemented with L-LDH and NAD was investigated. When LDH or NAD were added to the sucrose rinse, the pH fall was less marked than with rinsing with sucrose alone. Significant differences (p < 0.05) in mean minimum pH and cH areas were observed when sucrose rinses were supplemented with LDH, NAD or with a combination of LDH and NAD, when compared with sucrose alone. Plaque fluid concentrations of lactate and acetate significantly decreased with all rinses when compared with sucrose alone. Phosphate levels also decreased, although not significantly, when NAD was the sole supplement. No significant decreases were observed in succinate, formate or propionate concentrations. The results confirm the beneficial effect of LDH in reducing the accumulation of lactate in plaque following a sucrose rinse. The inclusion of NAD in the rinse demonstrated the role of extracellular hydrogen acceptors in the reaction. Further work is required to identify endogenous hydrogen receptors and to clarify the fate of pyruvate resulting from the oxidation of lactate before the potential role of LDH as a cariostatic agent can be fully understood.

Effects of dietary sucrose levels on pH fall and acid-anion profile in human dental plaque after a starch mouth-rinse.

The effects on the metabolism of starch by plaques formed in human subjects during periods of dietary sucrose limitation or supplementation were studied. High sucrose (HS) plaques showed lower resting pH and pH minima, and higher concentrations of lactate ion after the starch mouth-rinse than low sucrose (LS) plaques. Plaque samples incubated with starch solutions in vitro showed no differences in final pH or acid-anion concentrations between HS and LS diets. Plaque-amylase activity was higher on HS than LS diets. Thus starch was more acidogenic when consumed as part of a diet already rich in sucrose, but it is unclear whether this was a specific or non-specific effect of starch on plaque metabolism.

Human dental plaque pH, and the organic acid and free amino acid profiles in plaque fluid, after sucrose rinsing.

The relationship between these factors was studied in plaque and plaque fluid samples taken at intervals during the Stephan pH curve following a sucrose mouth rinse. Levels of lactate rose after the rinse, then fell during the pH recovery phase. Levels of acetate, propionate and phosphate fell after rinsing, then rose again. Amino acid concentrations also changed, with many showing a fall followed by a rise; others rising then falling; and some showing a more variable or complex pattern. In resting plaque fluid, only alanine, proline, glutamic acid, glycine and ammonia were present at concentrations above 1 mmol/l. Delta-aminovaleric acid was detected at levels below those that have been found in monkeys. Hydroxyproline and hydroxylysine were consistently detected, levels of arginine were generally low, and those of cystine consistently very low. The results may provide a basis for understanding the complex metabolic interrelations that occur in the course of the Stephan curve and which may reflect or produce the observed pH changes. They suggest that besides the amount of acid produced, the type of acid, buffering power and base production should be considered as determinants of plaque pH.

The validation of quantitative light-induced fluorescence to quantify acid erosion of human enamel.

OBJECTIVE: The purpose of this study was to validate the Quantitative light-induced fluorescence (QLF) device against transverse microradiography (TMR) with regard to the quantification of enamel erosion in vitro. DESIGN: Longitudinal in vitro. METHODS: Thirty previously extracted, caries free, human premolars were selected and prepared by gentle pumicing and coating in an acid-resistant nail-varnish save for an exposed window on the buccal surface. QLF baseline images were taken and the teeth then exposed to an erosive solution, 0.1% citric acid (pH 2.74). Teeth were removed at 30min intervals, air-dried and QLF images taken. At this time one tooth was randomly selected, removed from solution and sectioned through the lesion at three sites. The polished sample (100microm) was subjected to TMR and analysed for erosive mineral loss using proprietary software, with the DeltaZ values noted. QLF images were analysed by a blinded examiner with DeltaF and DeltaQ values recorded. Data were entered into SPSS and the correlation between the DeltaZ and DeltaF, and DeltaZ and DeltaQ values calculated. RESULTS: A wide range of erosive lesions was produced, with a steady increase in both DeltaZ and DeltaF over time; DeltaZ (24.0 (S.D. 1.2)-6114.3 (S.D. 1177.57)); DeltaF (1.8-11.2), DeltaQ (2.5-202.6). The results were scatter plotted and a regression line calculated. A positive correlation between DeltaZ and DeltaF of 0.91 was found, and for DeltaZ and DeltaQ; 0.87. CONCLUSIONS: The ability for QLF to detect and longitudinally monitor in vitro erosion has been shown. The strong positive correlation of DeltaF with DeltaZ suggests that percentage fluorescence loss as measured by QLF could be of great value in the development of a non-destructive, longitudinal tool for use in vitro, in situ and possibly in vivo.

Factors affecting the development of carious lesions in bovine teeth in vitro.

This study aimed to determine the effect of temperature, duration of exposure, position on enamel surface, and type of demineralization solution on the production of caries-like lesions in bovine enamel in vitro, and to establish the conditions for the formation of artificial caries in bovine enamel. Caries-like lesions were produced in incisal, middle and cervical sites on enamel samples, with either an acidified hydroxyethylcellulose gel system or a partially saturated acidic buffer solution at either 20 degrees C or 37 degrees C for 3, 4, or 5 days. Lesion variables (mineral loss/lesion depth) were quantified. Regular subsurface lesions were produced in all specimens in acidic buffer solution within 3 days at either temperature. In gel, caries-like lesions were produced in 62% of the specimens at 37 degrees C and in 49% at 20 degrees C, while the remaining specimens were either eroded or softened. Mineral loss and lesion depth were significantly greater with buffer than with gel, and with increased length of exposure in either solution. There were no significant differences in either variable with position or temperature in either solutions, though numerically both variables were greater at the cervical site, and at 37 degrees C in either solution. It was concluded that caries-like lesions can be consistently produced in bovine enamel with a partially saturated acidic buffer solution at 20 degrees C or 37 degrees C within 3 days.

The influence of xylitol and fluoride on dental erosion in vitro.

The aim was to determine the effect of xylitol, fluoride and xylitol/fluoride combined on the erosion of dental enamel by pure orange juice in vitro. Freshly extracted bovine incisors were sectioned vertically into four equal portions. Each portion was then coated with an acid-resistant nail varnish except for an enamel window on the labial surface of the tooth. These were then divided into four groups with each group containing one portion of each tooth selected randomly. Four erosive agents were prepared as follows: (A) pure orange juice only; pure orange juice plus either (B) xylitol (25% w/v) or (C) fluoride (0.5 parts/10(6)) or D) xylitol/fluoride (25% and 0.5 parts/10(6) respectively). Each group was assigned to one of the erosive agents and immersed six times daily for a period of 5 min on each occasion and stored in artificial saliva between exposures and for 12 hr overnight, for 24 days making a total of 12 hr of exposure to the assigned erosive agent. Sections were cut from each enamel specimen ground to a thickness of 80 microns and microradiographed. Mineral loss was quantified by a two-step image analysis. Mineral loss was significantly lower (p < 0.05) in group D (xylitol/fluoride) only when compared with group A (pure orange juice only). The numerical values of mineral loss could be ranked as follows: group D < group C < group B < group A. It was concluded that xylitol and fluoride have an additive effect in the reduction of dental erosion by pure orange juice in vitro.

Detection of in vitro demineralization adjacent to restorations using quantitative light induced fluorescence (QLF).

AIM: Quantitative light-induced fluorescence (QLF) is a technique for the detection, quantification, and longitudinal monitoring of early carious lesions. The technique is non-destructive and can be used in vivo. Using the natural fluorescence of teeth, and the loss of such fluorescence in demineralized enamel, QLF is a repeatable and valid optical caries monitor. Previously used in smooth and occlusal surfaces, the purpose of this pilot study was to determine if QLF could detect, and longitudinally monitor, demineralization adjacent to a range of restorative materials. METHODS: Fifteen previously extracted lower third molars were selected based upon the lack of any visible demineralization. A single burr hole was placed on the buccal surface and the cavity restored with amalgam, composite, compomer, glass ionomer or a temporary filling material. The buccal surface was then coated in an acid resistant nail varnish leaving an exposed area around the restoration and also a similar sized control region. The teeth had QLF images taken at baseline and were then subjected to a demineralizing buffer, further QLF images were subsequently taken at 72 and 144 h. Transverse microradiography was used to confirm the presence of early, subsurface lesions at the completion of the cycle (144 h). QLF images were analyzed by a single blinded examiner and values for change in radiance fluorescence were computed. These values were recorded as loss of radiance fluorescence loss integrated over area of lesion and expressed as DeltaQ. RESULTS: The appearance of each material under QLF and the change in fluorescence is described. Amalgam, glass ionomer and the temporary material all exhibited reduced fluorescence, while composite and compomer showed increased fluorescence, when compared with surrounding enamel. There was no change in fluorescence of the materials when subjected to experimental demineralizing conditions. Readings at 72 and 144 h demonstrated demineralization adjacent to the restorations and at the exposed control. Significant differences were detected between baseline, 72 and 144 h using ANOVA on all restorations with the exception of compomer where significance was noted between baseline and 144 h, p>0.05. CONCLUSIONS: This pilot study has demonstrated the ability for QLF to detect and monitor secondary caries. Analysis techniques should be based upon the subtraction of baseline DeltaQ scores from subsequent images. Further research is required to assess the ability of QLF to detect secondary lesions in vivo.