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PubMed1996

Evaluation of sodium fluorescein for quantitative diagnosis of root caries.

The diagnosis of root caries, in particular the judgment of the activity of a visually observed lesion, is difficult. Quantitative determination of lesion severity would allow the lesion to be monitored with time, so that an indication of lesion activity could be obtained. This paper describes a step in the development of a method that provides such a quantitative determination. Specifically, fluorescein sodium salt is used as a penetrating dye, the subject of study being the relationship between dye concentration and porosity in demineralized root dentin. Fourteen human third molars were demineralized in vitro (lactic acid CMC-gel, pH 5; in each of 6 groups for 4, 7, 11, 14, 18, and 21 days). Fluorescein sodium salt (0.2 g/L) was applied for 2 min. Thin slices (+/- 130 microns) were cut from the root surfaces without water cooling. The dye fluorescence radiance in the demineralized dentin was determined by means of a micro-Raman spectroscope and compared with the mineral loss profiles measured with transverse microradiography (TMR). The TMR data were corrected for the difference in measurement area between the two measurement systems. Corrected TMR profiles were compared with the corresponding fluorescence scans, showing linear correspondence. The correlation coefficient was r = 0.96. We conclude that, after uptake of fluorescein sodium salt for 2 min, the dye concentration in an artificially produced root-surface caries lesion is proportional to the amount of mineral lost from that lesion.

Dentin↗
PubMed1996

The effect of a fluoridated toothpaste on dentinal lesions in plaque-filled grooves: an intra-oral crossover study.

Recently, we introduced an intra-oral dentin demineralization model to evaluate the efficacy of a fluoridated toothpaste at plaque-retention sites with limited access to salivary components and fluoride. Usage of the fluoridated toothpaste reduced mineral loss from grooves in sound dentin by 15% compared with a non-fluoridated toothpaste (Lagerweij et al., 1997). In the current study, preformed dentinal lesions were subjected to similar toothpaste treatments. To improve the power of the model in discriminating between toothpaste groups, we changed the design from monadic to crossover to reduce the influence of variation between the participants. Two sets of specimens were placed in the partial prostheses of 29 participants who used a toothpaste containing either 1000 ppm fluoride or no fluoride during two consecutive six-week periods. Changes in mineral content were analyzed by transverse microradiography. The average mineral loss after the non-fluoride period was 1785 vol% x micron (SD 2399), and 1335 vol% x micron (SD 2039) after the fluoride period, a 25% inhibition in demineralization. Analysis of variance showed that the demineralization was significantly influenced not only by the type of treatment (p < 0.001 level), but also by the participant (p < 0.001), the width of the groove (p < 0.001), and the depth into the groove where changes in mineral content were measured (p < 0.017). Twenty-one percent of all the lesions showed remineralization. No significant difference in the extent or the occurrence of remineralization was found between the treatments.

Adult↗
PubMed1997

Remineralization of enamel subsurface lesions by casein phosphopeptide-stabilized calcium phosphate solutions.

Casein phosphopeptides (CPP) stabilize amorphous calcium phosphate (ACP), localize ACP in dental plaque, and are anticariogenic in animal and in situ human caries model. In this vitro study, CPP-stabilized calcium phosphate solutions were shown to remineralize subsurface lesions in human third-molar enamel. Solutions were used to examine the effect of CPP-calcium phosphate concentration on remineralization. Other solutions were used to examine the effect of increasing pH, which decreased the concentrations of free calcium and phosphate ions and increased the level of CPP-bound ACP. Although most of the remineralizing solutions were supersaturated with respect to the amorphous and crystalline calcium phosphate phases, the solutions were stabilized by the CPP such that spontaneous precipitation of calcium phosphate did not occur. After a ten-day remineralization period, enamel lesions were sectioned, subjected to microradiography, and the mineral content determined by microdensitometry. All solutions deposited mineral into the bodies of the lesions, with the 1.0% CPP-calcium phosphate (pH 7.0) solution replacing 63.9 +/- 20.1% of mineral lost at an averaged rate of 3.9 +/- 0.8 x 10(-8) mol hydroxyapatite/m2/s. The remineralizing capacity was greater for the solutions with the higher levels of CPP-stabilized free calcium and phosphate ions. Remineralization was not significantly correlated with either the CPP-bound ACP of the degrees of saturation for hydroxyapatite, octacalcium phosphate, or ACP. However, remineralization was significantly correlated with the degree of saturation for dicalcium phosphate dihydrate (CaHPO4.2H2O), but his was attributed to the significant correlation of remineralization with the activity gradients from the solution into the lesion of some calcium phosphate ions and ion pairs, in particular the neutral ion pair CaHPO4(0). The CPP, by stabilizing calcium phosphate in solution, maintain high-concentration gradients of calcium and phosphate ions and ion pairs into the subsurface lesion and thus effect high rates of enamel remineralization.

Calcium↗
PubMed1999

Kinetics of enamel demineralization in vitro.

Previously, we reported that the rate (R) of hydroxyapatite dissolution in acetic, lactic, and phosphoric acid solutions is a function of the degree of saturation with respect to the dissolving mineral, DS (defined as the ratio of the mean ionic activity product for hydroxyapatite [Ca5OH(PO4)3] in solution to its solubility product constant), and the sum of the acid activities (sumBiH) in solution: R = K(1-DS)m(sumBiH)n. The present study was undertaken to explore the general validity of this model in describing the kinetics of enamel demineralization. Thin sections of human enamel were exposed to partially saturated 0.1 mol/L lactic acid solutions, at two different DS levels, and at pH values of 4.3 to 6.0. Thin sections of human enamel were also exposed to solutions with four different concentrations of acetic and lactic acids (pH 4.3) with three different DS values and, at one DS value, to solutions of propionic acid. Mineral loss was monitored by quantitative microradiography. In solutions with pH values of 4.3 and 5.0, "lesions" were formed with well-defined surface layers, whereas, in solutions with pH 6.0, "lesions" were produced with no apparent surface layers. The formation of relatively intact surface layers was consistent with predicted phase transformations. Rates of mineral loss were found to be inversely proportional to both the degree of saturation with respect to enamel mineral, DS(En), and the pH of the solution and increased with increased activities of each organic acid, consistent with the proposed model. However, at the same DS(En) and acid activity, rates of demineralization were the same in the acetic and propionic acid solutions, whereas rates of demineralization in lactic acid were greater. It is suggested that specific interactions of acid species with enamel mineral may modify the rate of enamel demineralization. These in vitro findings suggest that relatively small differences in DS(En) values found in plaque fluid may result in very significant differences in the rate of enamel demineralization in vivo.

Acetates↗
PubMed1999

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.

Analysis of Variance↗
PubMed2002

Effect of perfusion with water on demineralization of human dentin in vitro.

Dentinal fluid rarely features in caries studies of dentin. The mutual effects of in vitro perfusion and dentin demineralization were investigated. The correlation between the remaining dentin thickness and demineralization was also analyzed. Buccal cervical dentin windows in human tooth segments were demineralized either with or without perfusion with water at 3.14 kPa. Transverse microradiography revealed that dentin perfusion reduced the amount of mineral loss from the lesions by 22 vol%; the reduction in lesion depth was 8%. Perfusion rate, which was measured throughout the demineralization process by means of a fluid transport model, did not change significantly. Lesions formed closer to the pulp exhibited increased mineral loss and lesion depth. In conclusion, dentinal fluid flow offers some protection against demineralization. For a better approximation of clinical reality, therefore, in vitro studies on dentinal caries should consider the effect of dentinal fluid flow.

Analysis of Variance↗
PubMed1997

Reproducibility of TMR for the determination of longitudinal mineral changes in dental hard tissues.

Transverse microradiography (TMR) of thin sections is the principal method for determining mineral changes in experimental lesions. Because it has become also the standard by which newly-developed methods are validated, demands on all steps of the procedure are high. Thin sections of enamel and dentin require high-precision cutting or lapping to preserve surface structures, but their different brittleness and elasticity preclude a uniform procedure. Image analysis permits the measurement of fine details at a micron scale, but because most studies are aimed at overall mineral changes, they are usually lost on broadening of the scan areas and averaging of experimental groups. Small errors in repositioning of the scan areas for repetitive measurements may yield distorted lesion profiles, while quantitative data such as the integrated mineral loss are hardly influenced. A major reason for "irreproducibility" of TMR are the different definitions of the lesion parameters as used by various research groups. We recommend that these definitions be agreed on if the full potential of TMR as an international standard is to be utilized.

Absorptiometry, Photon↗
PubMed1997

In vitro studies of laser fluorescence for detection and quantification of mineral loss from dental caries.

Laser fluorescence (LF) is thought to be a quantitative technique for the measurement of mineral loss from dental caries. These studies aimed to demonstrate the quantitative nature of LF, by means of transverse microradiography (TMR), in addition to the possible use of LF on small specimens suitable for in situ studies. Studies also aimed to determine the ability of LF to detect caries adjacent to amalgam restorations. The results demonstrated reasonable correlation between both the histological depth and mineral loss measured by TMR compared with the % change in mean fluorescence radiance measured by LF (r = 0.70 and 0.83, respectively). Studies with small specimens demonstrated that in vitro remineralization could be observed with LF for comparison of the demineralized specimen with an undemineralized reference. ANOVA showed significant mineral gain (p < 0.0001). The detection and quantification of mineral loss adjacent to amalgam restorations may have potential for the management of recurrent caries. Studies have demonstrated that LF can detect such mineral loss, and, with refinement of the image analysis system, LF was capable of detecting remineralization. Remineralization of such lesions is controversial, but it is suggested that LF may have a role in secondary caries management.

Analysis of Variance↗
PubMed2002

The effects of the solubility of artificial fissures on plaque pH.

Dissolution of the fissure walls may buffer acids formed in plaque and thus prevent the penetration of acids into the fissure. To test this, five volunteers wore dentin, enamel, and polyacrylate specimens with narrow grooves for 7 days to accumulate plaque. Temporal (pre- and post-glucose) and spatial (0-0.7 mm) pH profiles were recorded in the grooves in a flow-through reactor with pH microsensors. Mineral loss was assessed by transverse microradiography. We observed that resting pH did not differ among substrata. The median pH 1 hr post-glucose at the bottoms of dentin, enamel, and polyacrylate grooves was 6.7, 6.2, and 5.7, respectively (p < 0.01). On subject level, lesions formed in dentin correlated with pH changes in polyacrylate, where no buffering of acids due to mineral dissolution occurred. We conclude that fluoride-deficient tissue at the bottom of a fissure is at increased risk for caries, if acids are not buffered near the entrance to the fissure.

Acids↗
PubMed2003

Effect of smear layer on root demineralization adjacent to resin-modified glass ionomer.

The cariostatic effect of resin-modified glass ionomer (RMGI) on secondary root caries is well-documented. However, this beneficial effect may be dependent upon the mode of cavity surface treatment. To investigate this relationship, we studied 4 cavity surface treatments prior to the placement of RMGI: no treatment (None), polyacrylic acid (PAA), phosphoric acid (H(3)PO(4)), and Scotchbond Multi-Purpose adhesive (SMP) as a control. Specimens were aged for two weeks in synthetic saliva, thermocycled, and subjected to an artificial caries challenge (pH 4.4). Polarized light microscopy (PLM) and microradiography (MRG) showed significantly less demineralization with the H(3)PO(4) cavity surface treatment as revealed by ANOVA and Tukey's multiple comparisons (p < or = 0.05). Dentin fluoride profiles determined by electron probe microanalysis (EPMA) supported PLM and MRG findings. It may be concluded that removal of the smear layer with phosphoric acid provides significantly enhanced resistance to secondary root caries formation adjacent to RMGI restorations.

Acid Etching, Dental↗
PubMed2004

Effect of dentinal fluid composition on dentin demineralization in vitro.

Dentin demineralization is reduced by perfusion with water. We hypothesized that a simulated dentinal fluid (SDF) that contains albumin, in addition to electrolytes, would be more effective in reducing dentin demineralization than water alone, and this effect would increase with increasing flow rate of SDF. Perfusion rate in tooth segments that carried buccal cervical dentin windows was measured in a fluid transport set-up. These windows were then demineralized under perfusion with water, or SDF at 1.47 kPa for 31 days. We analyzed integrated mineral loss and lesion depth with the use of transverse microradiography (TMR), which revealed that 38% more mineral dissolved from dentin lesions perfused with water than from those perfused with SDF. The former were also 18% deeper. Flow rate of dentinal fluid showed no correlation with demineralization. We concluded that composition of dentinal fluid is an important determinant of the rate of lesion formation and progression in dentin.

Albumins↗
PubMed2006

The effect of copper on demineralization of dental enamel.

Previous studies have concluded that copper might inhibit enamel demineralization in vitro. Our aim was to assess the effect of copper (Cu2+), with and without amine fluoride, on human dental enamel under cariogenic challenge in situ. In a double-blind randomized four-leg crossover trial, 14 individuals wore a removable appliance containing 2 enamel slabs, 1 containing an artificial caries lesion. During each leg, the appliance was exposed twice daily to one of the test solutions: 1.25 mM CuSO4, amine fluoride (250 ppm F), copper and amine fluoride combined, or a placebo (water). A cariogenic challenge was provided in all cases by 5 daily exposures to 10% sucrose. Slabs were assessed before and after 21 days' exposure by Knoop microhardness and transverse microradiography. Significantly less demineralization was observed with Cu2+ and fluoride in combination than with fluoride treatment alone (p < 0.05), whereas copper alone had no significant protective effect.

Adult↗
PubMed2003

Contribution of raloxifene and calcium and vitamin D3 supplementation to the increase of the degree of mineralization of bone in postmenopausal women.

Raloxifene has been shown to increase bone mineral density and reduce the risk of vertebral fracture in postmenopausal women with osteoporosis. In this study, we report the results of the first prospective longitudinal study to evaluate the mean degree of mineralization of bone (MDMB) in a group of patients enrolled in the Multiple Outcomes of Raloxifene Evaluation trial. Patients were randomly assigned to one of three treatment groups: placebo (n = 24), raloxifene 60 mg/d (RLX60; n = 22), or raloxifene 120 mg/d (RLX120; n = 18). All patients received daily calcium (500 mg) and vitamin D(3) (400-600 IU) supplementation for the duration of the study. Iliac crest biopsies were taken at baseline and after 2 yr of treatment. Quantitative microradiography was used to analyze the biopsy specimens and revealed a statistically significant (P < 0.05) mean percentage increase in total MDMB of 7.0, 5.3, and 5% for RLX60-, RLX120-, and placebo-treated patients, respectively, compared with baseline. Raloxifene treatment was found to shift the distribution of total bone mineral to higher values of MDMB (RLX60, 29%; RLX120, 8%) with greater heterogeneity, compared with placebo. The profile of MDMB observed in biopsies after treatment with placebo and raloxifene, compared with baseline, closely resembles physiological premenopausal bone.

Aged↗
PubMed1996

Microcalcification clustering parameters in breast disease: a morphometric analysis of radiographs of excision specimens.

X-ray microradiography of surgically excised breast specimens offers the possibility of morphological characterization of calcifications. When combined with digital imaging techniques there exists added potential for obtaining valuable basic quantitative morphometric information regarding differences between microcalcifications in tissues exhibiting evidence of fibrocystic change, benign and malignant tumours. A total of 157 excised breast specimens from 84 patients were microradiographed using a Softex Super Soft X-ray unit and Kodak AA high resolution industrial film. A Quantimet 570C image analysis system was used to digitize and analyse the microradiographs. Of the 157 microradiographs, 51 (from 30 patients) revealed microcalcification clusters. The existence of significant differences between the three identified categories of tissue were indicated by clustering parameters. These included the number of particles per cluster, area of clusters, maximum distance to nearest neighbour, and geometric mean distance to nearest neighbour. The distribution pattern index (DPI), another of the clustering parameters used in this study, has been observed to be a particularly powerful discriminator. The value for fibrocystic change was found to be significantly smaller (0.514) than that for benign tumour (0.796) whilst that for benign tumour was observed to be significantly larger than that for malignant tumour (0.604) at a p-value of less than 0.05 (Kruskal-Wallis one-way analysis of variance).

Adolescent↗
PubMed1976

Protection from stress in bone and its effects. Experiments with stainless steel and plastic plates in dogs.

In a preliminary experiment the paired radii and femora of dogs were tested for bone mineral mass and mechanical properties including the load at break, the ultimate bending strength and the modulus of elasticity; symmetry was observed for most of the parameters determined. The influence of the elasticity of materials used for the internal splintage of bone and its relationship to bone remodeling were then investigated for stainless steel and plastic plates applied to the femora of dogs. A significant decrease in bone mineral mass per centimetre length of bone and in mechanical properties was demonstrated for the femora plated with steel, and microradiography showed that this was due to massive endosteal resorption. A model for determining the influence of protection from stress in bone is presented.

Animals↗
PubMed2005

Long-term strontium ranelate administration in monkeys preserves characteristics of bone mineral crystals and degree of mineralization of bone.

UNLABELLED: In monkeys, long-term strontium ranelate administration results in a dose-dependent bone strontium uptake (mainly into newly formed bone) that preserves the degree of mineralization of bone and the bone mineral at the crystal level, showing its safety at bone mineral level. INTRODUCTION: Strontium ranelate simultaneously increases bone formation and decreases bone resorption, leading to prevention of bone loss and increase in bone mass and bone strength in normal and ovariectomized rats. This study investigated the interactions of stable strontium (Sr) with bone mineral in monkeys after long-term strontium ranelate treatment and after a period of treatment withdrawal. MATERIALS AND METHODS: Iliac bone was obtained from untreated monkeys, monkeys at the end of a 52-week strontium ranelate administration (200, 500, 1250 mg/kg/day orally), and in parallel groups 10 weeks after the end of strontium ranelate administration (same three doses; n = 3-7). Sr uptake and distribution in bone mineral were quantified by X-ray microanalysis, changes at the crystal level by X-ray diffraction, and the degree of mineralization of bone (DMB) by quantitative microradiography. RESULTS: After strontium ranelate administration, dose-dependent Sr uptake occurred into cortical and cancellous bone, with higher content (1.6 times) in new than in old bone. This Sr uptake decreased (50%) 10 weeks after treatment withdrawal; the decrease occurred almost exclusively in new bone. At the end of strontium ranelate treatment and after its withdrawal, a preservation of crystal characteristics was observed, suggesting that Sr was only faintly linked to crystals by ionic substitution and of DMB. CONCLUSIONS: These results show the absence of a deleterious effect of long-term strontium ranelate treatment on bone mineralization, confirming the histomorphometric observations made in postmenopausal osteoporotic women treated with strontium ranelate.

Animals↗
PubMed2001

Distribution of intracortical porosity in human midfemoral cortex by age and gender.

The purpose of this study was to describe the age-specific distribution of midfemoral intracortical porosity throughout the cortical width in males and females. Microradiography and an automated image analysis system were used to study midfemoral cortical bone specimens from 163 white people, including 77 males and 86 females, in a recent anthropological collection covering a broad age range. In each specimen, porosity (percentage of the cortical bone area occupied by pores), pore number, and pore size were measured throughout the entire cortex and in three cortical subregions of equal width labeled the periosteal, midcortical, and endosteal subregions. For each gender, relationships linking age to porosity, pore number, and mean pore size were assessed using regression analysis. In addition, age- and site-related changes in these three variables were tested for significance using two-way analysis of variance (ANOVA). Age explained 52% of the porosity variance in females and 13.5% in males. In each gender, there were significant age- and site-related differences in porosity, pore number, and pore size. In adults aged 60 years or younger, both pore size and pore number increased with increasing age, whereas in adults older than 60 years, pore size continued to increase but pore number decreased. In males, the age-related changes in pore size and pore number were proportionally similar in the three cortical subregions. In females, in contrast, the changes predominated in the endosteal subregion and resulted in significant cortical thinning.

Adolescent↗
PubMed2004

Remineralization of carious dentin. I: in vitro microradiographic study in human teeth capped with calcium hydroxide.

The objective of this in vitro study was to evaluate a possible remineralization of human carious dentin by calcium hydroxide. Thirty-nine freshly extracted human permanent and deciduous carious teeth were split into two halves. One half was used as control and the other as experimental. In the latter, a cavity was prepared and the remaining bottom layer of demineralized dentin capped with chemically pure calcium hydroxide. The experimental samples were stored at 37 masculine C. Time intervals were 2, 4, 6, 8, 10 and 12 weeks. All tooth halves were sawed and ground to plano-parallel sections with a thickness varying from 75 to 117 microm. Qualitative microradiographs showed a qualitative increase in radiopacity of the calcium hydroxide treated samples. Quantitative microradiography showed a statistically significant increase in total mineral content in the experimental samples compared to the control samples. These results indicate an in vitro remineralization of carious dentin by calcium hydroxide.

Calcium Hydroxide↗