Remineralization effectiveness of the PAMAM dendrimer with different terminal groups on artificial initial enamel caries in vitro

ObjectiveDisruption of the demineralization–remineralization balance could trigger the development of dental caries, making it challenging for enamel to “self-heal”. Thus, extrinsic assistance is needed to restore enamel lesions and stop undermining progression. The aim of this study was to investigate enamel remineralization in a simulated oral environment via poly (amino amine) (PAMAM) dendrimers quantitatively.MethodsBovine enamel specimens were shaken in demineralization solution (pH 4.5, 37 °C, 50 rpm/min) for 72 h to create initial enamel carious lesions. The subsurface-demineralized specimens were then divided into four groups: enamel treated with PAMAM-NH₂, enamel treated with PAMAM−COOH, enamel treated with PAMAM−OH, and enamel treated with deionized water. The treated specimens underwent subsequent 12-day pH cycling. Enamel blocks were analyzed by transverse microradiography (TMR), surface microhardness testing and scanning electron microscopy (SEM) before and after demineralization and pH cycling.ResultsGroups treated with PAMAM dendrimers showed lower lesion depth and less mineral loss, attained more vertical-section surface microhardness recovery, and adsorbed more mineral deposits (p < 0.05). The enamel lesion remineralization values of PAMAM-NH₂, PAMAM-COOH, and PAMAM-OH groups were 76.42 ± 3.32%, 60.07 ± 5.92% and 54.52 ± 7.81%, respectively.SignificanceIn conclusion, PAMAM with different terminal groups could induce enamel remineralization, among which PAMAM-NH₂ showed the most prominent competence, followed by PAMAM-COOH and PAMAM-OH, in that order.     

The effect of xylitol and fluoride on remineralization for primary tooth enamel caries in vitro

The effect of fluoride and xylitol on remineralization at the early stage of the enamel caries in primary tooth was studied. The samples were divided into four groups (control, 10% xylitol, 950 ppm NaF and 10% xylitol+950 ppm NaF) and analyzed by the using single thin section method and pHcycling model in vitro. The remineralizing ratio were control –8.9%, xylitol –0.4%, NaF 8.3% and xylitol + NaF 32.4%, respectively. Xylitol+NaF group particularly showed significantly smaller ΔZ value compared with 0 days (P<0.05). Therefore we assume that the effect of xylitol and fluoride are additive. We concluded that xylitol and fluoride treatment to the tooth enamel may be an effective caries-preventive measure in both primary and permanent tooth enamel.     

The effect of surface defects in early caries assessment using quantitative light-induced fluorescence (QLF) and micro-digital-photography (MDP)

ObjectivesThe purpose of this study was to consider the impact of surface defects on quantitative light-induced fluorescence (QLF) and micro-digital-photography (MDP) measures, in relationship to lesion depth.MethodsSimulated enamel carious lesions were developed on 45 extracted human teeth. Images of each tooth were captured with both QLF and MDP. The teeth were sectioned and lesion depth was measured with polarized light microscopy (PLM). Pearson correlations were computed using data from the 27 lesions which did not have surface loss, and then separately based upon the 18 lesions which did display surface loss. MDP variables ΔR and ΔX measure reflected light, whereas QLF variables ΔF and ΔQ measure fluorescence.ResultsA strong correlation was identified between lesion depth and ΔF (r = ?0.765, p < 0.0001), and ΔQ (r = ?0.827, p < 0.0001) on intact lesions while a weak but suggestive, although non-significant, correlation was identified between average lesion depth and ΔR (r = 0.369, p = 0.059) and ΔX (r = 0.595, p = 0.0011). However, the corresponding correlation was not statistically significant, when lesions with surface loss were considered for QLF and MDP measures.ConclusionsQLF measures ΔF and ΔQ were strongly correlated with lesion depth in lab-simulated lesions with no surface loss, but not among lesions with surface defects. The two MDP-associated measures, ΔR and ΔX, could not be said to differ significantly when lesions with and without surface defects were compared with lesion depth. Because intact lesions can be remineralized, accurate assessment of their status is imperative for caries treatment.Clinical significanceDental caries is still widely prevalent today. We now know that with early stage detection, remineralization can be accomplished. Being able to identify dental caries in its reversible stage (before physical surface loss) is paramount for the clinician to be able to treat the disease non-invasively.     

Optimization of calcium concentration of saliva with phosphoryl oligosaccharides of calcium (POs-Ca) for enamel remineralization in vitro

ObjectivePhosphoryl oligosaccharides of calcium (POs-Ca) are highly soluble calcium source made from potato starch. The aim of this study was to investigate the optimal concentrations of POs-Ca for the remineralization of subsurface enamel lesions in vitro.DesignDemineralized bovine enamel slabs (n = 5) were remineralized in vitro for 24 h at 37 °C with artificial saliva (AS) containing 0–0.74% POs-Ca to adjust the Ca/P ratio to 0.4–3.0, then sectioned and analysed by transversal microradiography (TMR). The data were analysed by Scheffe's post hoc test. The Ca/P ratio with most remineralization was used to investigate the effect of calcium on enamel remineralization (n = 11). The demineralized slabs were treated with AS with calcium-chloride- (CaCl₂-) or POs-Ca with an identical calcium content, and sectioned for TMR and wide-angle X-ray diffraction (WAXRD) analyses to evaluate the local changes in hydroxyapatite (HAp) crystal content. The data were analysed using the Mann–Whitney U-test.ResultsThe highest mineral recovery rate resulted from addition of POs-Ca to adjust the Ca/P to 1.67. At this ratio, the mineral recovery rate for AS containing POs-Ca (24.2 ± 7.4%) was significantly higher than that for AS containing CaCl₂ (12.5 ± 11.3%) (mean ± SD, p < 0.05). The recovery rate of HAp crystallites for AS containing POs-Ca (35.7 ± 10.9%) was also significantly higher than that for AS containing CaCl₂ (23.1 ± 13.5%) (p < 0.05). The restored crystallites were oriented in the same directions as in sound enamel.ConclusionsPOs-Ca effectively enhances enamel remineralization with ordered HAp at a Ca/P ratio of 1.67.     

Randomized clinical trial on effectiveness of silver diamine fluoride and glass ionomer in arresting dentine caries in preschool children

ObjectiveTo compare the effectiveness of annual topical application of silver diamine fluoride (SDF) solution, semi-annual topical application of SDF solution, and annual application of a flowable high fluoride-releasing glass ionomer in arresting active dentine caries in primary teeth.MethodsA total of 212 children, aged 3–4 years, were randomly allocated to one of three groups for treatment of carious dentine cavities in their primary teeth: Gp1-annual application of SDF, Gp2-semi-annual application of SDF, and Gp3-annual application of glass ionomer. Follow-up examinations were carried out every six months to assess whether the treated caries lesions had become arrested.ResultsAfter 24 months, 181 (85%) children remained in the study. The caries arrest rates were 79%, 91% and 82% for Gp1, Gp2 and Gp3, respectively (p = 0.007). In the logistic regression model using GEE to adjust for clustering effect, higher caries arrest rates were found in lesions treated in Gp2 (OR = 2.98, p = 0.007), those in anterior teeth (OR = 5.55, p < 0.001), and those in buccal/lingual smooth surfaces (OR = 15.6, p = 0.004).ConclusionAnnual application of either SDF solution or high fluoride-releasing glass ionomer can arrest active dentine caries. Increasing the frequency of application to every 6 months can increase the caries arrest rate of SDF application.Clinical significanceArrest of active dentine caries in primary teeth by topical application of SDF solution can be enhanced by increasing the frequency of application from annually to every 6 months, whereas annual paint-on of a flowable glass ionomer can also arrest active dentine caries and may provide a more aesthetic outcome.     

Fabrication and characterization of remineralizing dental composites containing calcium type pre-reacted glass-ionomer (PRG-Ca) fillers

ObjectiveTo fabricate and characterize dental composites with calcium type pre-reacted glass-ionomer (PRG-Ca) fillers.MethodsPRG-Ca fillers were prepared by the reaction of calcium fluoroaluminosilicate glass with polyacrylic acid. Seven dental composites were produced from the same organic matrix (70/30 wt% Bis-GMA/TEGDMA), with partial replacement of barium borosilicate (BaBSi) fillers (60 wt%) by PRG-Ca fillers (wt%): E0 (0) – control, E1 (10), E2 (20), E3 (30), E4 (40), E5 (50) and E6 (60). Enamel remineralization was evaluated in caries-like enamel lesions induced by S. mutans biofilm using micro-CT. The following properties were characterized: degree of conversion (DC%), roughness (Ra), Knoop hardness (KHN), flexural strength (FS), flexural modulus (FM), water sorption (W_sp), water solubility (W_sl), and translucency (TP). Data were analyzed to one-way ANOVA and Tukey’s HSD test (α = 0.05).ResultsAll composites with PRG-Ca induced enamel remineralization. E0 and E1 presented similar and highest DC% than E2 = E3 = E4 = E5 = E6. Ra and KHN were not influenced by PRG-Ca fillers (p < 0.05). The higher the content of PRG-Ca, the lower FS, FM and TP (p < 0.05). W_sp increased linearly with the content of PRG-Ca fillers (p < 0.05). E6 presented the highest W_sl (p < 0.05), while the W_sl of the other composites were not different from each other (p > 0.05).SignificanceIncorporation of 10–40 wt.% of PRG-Ca fillers endowed remineralizing potential to dental composites without jeopardizing the overall behavior of their physicochemical properties. Dental composites with PRG-Ca fillers seems to be a good alternative for reinforcing the enamel against caries development.     

Sodium fluoride effect on erosion–abrasion under hyposalivatory simulating conditions

ObjectivesTo investigate the effect of fluoride (0, 275 and 1250 ppm F; NaF) in combination with normal and low salivary flow rates on enamel surface loss and fluoride uptake using an erosion–remineralization–abrasion cycling model.DesignEnamel specimens were randomly assigned to 6 experimental groups (n = 8). Specimens were individually placed in custom made devices, creating a sealed chamber on the enamel surface, connected to a peristaltic pump. Citric acid was injected into the chamber for 2 min followed by artificial saliva at 0.5 (normal flow) or 0.05 (low flow) ml/min, for 60 min. This cycle was repeated 4×/day, for 5 days. Toothbrushing with abrasive suspensions containing fluoride was performed for 2 min (15 s of actual brushing) 2×/day. Surface loss was measured by optical profilometry. KOH-soluble fluoride and enamel fluoride uptake were determined after the cycling phase. Data were analysed by two-way ANOVA.ResultsNo significant interactions between fluoride concentration and salivary flow were observed for any tested variable. Low caused more surface loss than normal flow rate (p < 0.01). At both flow rates, surface loss for 0 was higher than for 275, which did not differ from 1250 ppm F. KOH-soluble and structurally-bound enamel fluoride uptake were significantly different between fluoride concentrations with 1250 > 275 > 0 ppm F (p < 0.01).ConclusionsSodium fluoride reduced enamel erosion/abrasion, although no additional protection was provided by the higher concentration. Higher erosion progression was observed in low salivary flow rates. Fluoride was not able to compensate for the differences in surface loss between flow rates.     

Effect of application sequence of fluoride and CPP-ACP on remineralization of white spot lesions in primary teeth: An in-vitro study

ObjectiveTo explore how application sequence of casein phosphopeptide-amorphous calcium phosphate (CPP-ACP) and fluoride influences remineralization of enamel white spot lesions (WSL) in primary teeth.DesignIn this in-vitro study, artificial WSLs were created in 130 primary teeth. Teeth were divided into 4 groups (n = 27) and a control group (n = 22) and exposed to one of the following remineralization regimens for 10 weeks: Group-1; 500 ppm fluoride dentifrice; Group-2; 10% w/v CPP-ACP; Group-3; fluoride applied first, then CPP-ACP; Group-4; CPP-ACP applied first, then fluoride, and Group-5 was control. All groups were kept in a remineralizing solution. Mineral changes (ΔF) were quantified weekly using quantitative light-induced fluorescence. Statistical analysis was done using Statistical Package for the Social Sciences (SPSS version 20.0).ResultsRemineralization occurred in all groups to different degrees; changes from baseline were significant in groups 1–4 (P ≤ 0.05). Group-4 showed the earliest significant remineralization (after 2 weeks) among groups, (P < 0.001). Group-4 showed maximum changes in ΔF among groups; however, only differences with Groups 1 and 5 were significant (P < 0.05 and P < 0.01, respectively). Group-3 showed better remineralization than Groups 1, 2 and 5; however, the difference was only significant with Group-5 (P < 0.001). There were no significant differences between Group 1and 2, however, only Group 2 showed better remineralization than Group 5, (P < 0.01).ConclusionCombined treatment with CPP-ACP followed by fluoride exhibited the best remineralization of white spot lesions in primary teeth in this study. Combined treatment with fluoride followed by CPP-ACP showed a tendency towards better remineralization than fluoride or CPP-ACP alone.     

In vitro enamel erosion and abrasion-inhibiting effect of different fluoride varnishes

ObjectiveTo investigate the erosion and abrasion inhibiting effect of CPP-ACP/NaF and xylitol/NaF varnishes.MethodsBovine enamel samples (n = 40) were exposed to the following treatments (n = 10): NaF varnish (Duraphat^®, positive control); CPP-ACP/NaF varnish (MI varnishTM); xylitol/NaF (Profluorid^®) or distilled and deionized water (MilliQ^®, negative control). The samples were submitted for 3 days to 4 cycles/day of erosion (5 min in Sprite Zero) and 2 cycles of abrasion/day after the first and last erosive challenge, with a toothbrush machine and slurries of a placebo toothpaste for 15 s (50 strokes/s). Among the cycles and after the last daily cycle, the specimens remained in artificial saliva. The change in the enamel surface was evaluated by using 3D non-contact optical profilometry with surface roughness (Ra and Sa values) and tooth structure loss (TSL) measurements. Scanning electron microscopy (SEM) assessed the enamel topographic characteristics. Differences in the Ra, Sa and TSL among treatments were tested using one-way ANOVA followed by the Tukey test.ResultsAll varnishes promoted better results for Ra and Sa values than the negative control (p = 0.0001), without difference among them (p > 0.05). However, CPP-ACP/NaF varnish stimulated fewer TSL (7.09 ± 0.70 μm) compared to NaF varnish (10.33 ± 1.36 μm, p = 0.002), xylitol/NaF varnish (9.96 ± 0.41 μm, p = 0.007) and the negative control (18.38 ± 3.32 μm, p = 0.0001).ConclusionA single-application of fluoride topical varnishes was effective in reducing enamel wear. The CPP-ACP/NaF varnish had the best effect against enamel loss from an erosion-abrasion challenge.     

Nano-calcium phosphate and dimethylaminohexadecyl methacrylate adhesive for dentin remineralization in a biofilm-challenged environment

ObjectiveDentin remineralization at the bonded interface would protect it from external risk factors, therefore, would enhance the longevity of restoration and combat secondary caries. Dental biofilm, as one of the critical biological factors in caries formation, should not be neglected in the assessment of caries preventive agents. In this work, the remineralization effectiveness of demineralized human dentin in a multi-species dental biofilm environment via an adhesive containing nanoparticles of amorphous calcium phosphate (NACP) and dimethylaminohexadecyl methacrylate (DMAHDM) was investigated.MethodsDentin demineralization was promoted by subjecting samples to a three-species acidic biofilm containing Streptococcus mutans, Streptococcus sanguinis, Streptococcus gordonii for 24 h. Samples were divided into a control group, a DMAHDM adhesive group, an NACP group, and an NACP + DMAHDM adhesive group. A bonded model containing a control-bonded group, a DMAHDM-bonded group, an NACP-bonded group, and an NACP + DMAHDM-bonded group was also included in this study. All samples were subjected to a remineralization protocol consisting of 4-h exposure per 24-h period in brain heart infusion broth plus 1% sucrose (BHIS) followed by immersion in artificial saliva for the remaining period. The pH of BHIS after 4-h immersion was measured every other day. After 14 days, the biofilm was assessed for colony-forming unit (CFU) count, lactic acid production, live/dead staining, and calcium and phosphate content. The mineral changes in the demineralized dentin samples were analyzed by transverse microradiography.ResultsThe in vitro experiment results showed that the NACP + DMAHDM adhesive effectively achieved acid neutralization, decreased biofilm colony-forming unit (CFU) count, decreased biofilm lactic acid production, and increased biofilm calcium and phosphate content. The NACP + DMAHDM adhesive group had higher remineralization value than the NACP or DMAHDM alone adhesive group.SignificanceThe NACP + DMAHDM adhesive was effective in remineralizing dentin lesion in a biofilm model. It is promising to use NACP + DMAHDM adhesive to protect bonded interface, inhibit secondary caries, and prolong the longevity of restoration.     

Mineralization-defects are comparable in fluorotic impacted human teeth and fluorotic mouse incisors

ObjectiveFluoride excess of 0.05–0.07 mg F/kg bw/day in water or food additives like salt is the principal cause of endemic dental fluorosis. How fluoride causes these defects is not clear yet. Recent studies in rodents suggest that development of enamel fluorosis is associated with insufficient neutralization of protons released during the formation of hypermineralized lines.DesignHere we examined whether hypermineralization could also be assessed by MicroCT in developing molar enamel of humans exposed to fluoride.ResultMicro-CT analysis of hypomineralized enamel from human fluorotic molars graded by the Thylstrup–Fejerskov (TF) Index as III–IV showed weak hypermineralized lines and hypermineralized patches not seen in TF-I/II grade enamel. The mesio-distal sides of these molar teeth were significantly smaller (∼18%, p = 0.02) than in TF-I/II teeth.ConclusionThe patterns of changes observed in human fluorotic teeth were similar to those in fluorotic rodent incisors. The data are consistent with the hypothesis that also in developing human teeth fluoride-stimulated local acidification of enamel could be a mechanism for developing fluorotic enamel.     

Effects of heat-inactivated Bifidobacterium BB12 on cariogenicity of Streptococcus mutans in vitro

ObjectiveSince some probiotic bacteria are cariogenic themselves, their suitability for caries management is questionable. Inactivated bacteria or their supernatants have been found to exert probiotic effects, whilst having several advantages compared with living bacteria. We hypothesized that viable and heat-inactivated Bifidobacterium animalis BB12 reduces the cariogenicity of Streptococcus mutans (SM) in vitro.DesignWe assessed mono- and mixed species biofilms of SM and viable or heat-inactivated BB12. Biofilms were grown in a continuous-culture-system under cariogenic conditions on smooth proximal enamel or cavitated dentine. For each of eight experimental subsets (4 biofilms × 2 hard-tissue conditions), a total of 32 specimens was used. After 10 days, bacterial numbers of 12 biofilms per group were analysed, and all specimens submitted to transversal microradiography.ResultsMineral loss was higher in cavitated dentine than smooth enamel for all biofilms (p < 0.001, t-test). BB12-monospecies biofilms induced significantly less mineral loss than SM in both enamel (p < 0.05) and dentine (p < 0.001). Viable BB12 did not significantly reduce cariogenicity of SM (p > 0.05), whilst heat-inactivated BB12 decreased cariogenicity of SM in dentinal cavities (p < 0.01). Bacterial numbers were higher on dentine than enamel (p < 0.05), but not significantly influenced by biofilm species (p > 0.05).ConclusionsHeat-inactivated BB12 reduced the cariogenicity of SM in dentinal cavities in vitro. Inactivated probiotics might be suitable for caries control.     

Graded changes in enamel component volumes resulted from a short tooth bleaching procedure

AimTo test the hypothesis that changes in enamel component volumes (mineral, organic, and water volumes, and permeability) are graded from outer to inner enamel after a short bleaching procedure.Materials and methodsExtracted unerupted human third molars had half of their crowns bleached (single bleaching session, 3 × 15 min), and tooth shade changes in bleached parts were analyzed with a spectrophotometer. Ground sections were prepared, component volumes and permeability were quantified at histological points located at varying distances from the enamel surface (n = 10 points/location), representing conditions before and after bleaching.ResultsTooth shade changes were significant (p < 0.001; 95% CI = −1/−8; power = 99%), and most of the enamel layer was unaffected after bleaching, except at the outer layers. Multiple analysis of covariances revealed that most of the variance of the change in enamel composition after bleaching was explained by the combination of the set of types of component volume (in decreasing order of relevance: mineral loss, organic gain, water gain, and decrease in permeability) with the set of distances from the enamel surface (graded from the enamel surface inward) (canonical R² = 0.97; p < 0.0001; power > 99%).ConclusionsChanges in enamel composition after a short bleaching procedure followed a gradient within component volumes (mineral loss > organic gain > water gain > decrease in permeability) and decreased from the enamel surface inward.     

Effect of iron on enamel demineralization and remineralization in vitro

ObjectiveTo evaluate the effect of ferrous sulphate on enamel demineralization and remineralization, using pH-cycling models.DesignFifty blocks were selected by their initial surface hardness and subjected to a pH-cycling demineralization process. Artificially demineralized lesions were produced in 60 blocks; out of these blocks, the surface hardness of 50 blocks and the cross-sectional hardness of 10 blocks were determined. The 50 blocks were then subjected to a remineralization pH-cycling process. Treatments were carried out using ferrous sulphate solutions of different concentrations (0.333, 0.840, 18.0, and 70.0 μg Fe/mL) and a control group (deionized water). The final surface hardness (SH₂) was determined, and the integrated subsurface hardness (ΔKHN) was calculated. The enamel blocks were analysed for fluoride, calcium, phosphorus, and iron. The obtained data were distributed heterogeneously and were analysed using the Kruskal–Wallis test (p < 0.05).ResultsIn demineralization pH cycling, the group treated with the 18.0 μg Fe/mL solution had higher secondary surface hardness and lower integrated subsurface hardness (ΔKHN) than the other groups. In remineralization pH cycling, the control group showed the lowest value of ΔKHN. A decline in Ca and P concentration was observed when the Fe concentration increased (p < 0.05). There was no significant difference in the F concentration (p > 0.05) and an increase in Fe concentration (p < 0.05) in the enamel was observed when the Fe concentration increased in both the demineralization and remineralization experiments.ConclusionThe results suggest that iron reduces demineralization but does not allow remineralization to occur.     

In vivo comparison between measurement from two fluorescence-based devices of occlusal and smooth surface caries in primary and permanent teeth

Fluorescence-based devices are adjunct tools for caries detection. Both DIAGNOdent 2095 (LF) and DIAGNOdent 2190 (LF pen) were developed for this application. The purpose of this in vivo study was to compare the relationship between the measurements obtained using the LF and LF pen devices in primary and permanent teeth. The same sample of occlusal and smooth surfaces caries were evaluated by one examiner using both LF and LF pen devices in children. For occlusal caries, measurements were made at 51 sites in primary teeth, and at 83 sites in permanent teeth. For smooth surface caries, measurements were made at 63 sites in primary teeth, and 163 sites in permanent teeth. The values obtained with the LF pen for occlusal caries in primary and permanent teeth (17.6 ± 2.14, 20.6 ± 1.40, respectively) were significantly higher than those obtained with the LF (9.7 ± 1.24, 11.3 ± 0.78). For caries on smooth surfaces, the values obtained with the LF pen for primary and permanent teeth (8.2 ± 1.22, 7.2 ± 0.45) were higher than those obtained with the LF (4.5 ± 0.57, 3.3 ± 0.23). For both occlusal and smooth surface caries, the values were significantly different between the two devices (P < 0.01). The Spearman's correlation coefficients comparing the LF and LF pen values for primary and permanent teeth were Rs = 0.87 and 0.86 for occlusal caries and 0.80 and 0.74 for smooth surface caries, respectively. Values from both devices showed high correlation coefficients, indicating similar principle of function. However, the actual values were significantly different, which indicated that the values obtained with the two devices did not agree. The LF pen should be used with caution in patients for whom the LF is already being used to manage caries.     

The effect of magnesium hydroxide-containing dentifrice using an extrinsic and intrinsic erosion cycling model

ObjectiveTo evaluate, in vitro, the effect of Mg(OH)₂ dentifrice, and the influence of the number of experimental days, on the extrinsic (citric acid –CA) and intrinsic (hydrochloric acid –HCl) enamel erosion models.DesignHuman enamel slabs were selected according to surface hardness and randomly assigned to 3 groups (n = 9) as follows: non-fluoridated (negative control), NaF (1450 ppm F- positive control) and Mg(OH)₂ (2%) dentifrices. The slabs were daily submitted to a 2-h period of pellicle formation and, over a period of 5 days, submitted to cycles (3×/day) of erosive challenge (CA 0.05 M, pH = 3.75 or HCl 0.01 M, pH = 2 for 30 s), treatment (1 min −1:3 w/w of dentifrice/distilled water) and remineralization (artificial saliva/120 min). Enamel changes were determined by surface hardness loss (SHL) for each day and mechanical profilometry analysis. Data were analyzed by two-way ANOVA followed by Tukey’s test to % SHL and one-way ANOVA to profilometry (p < 0.05).ResultsThe number of experimental days influenced the erosion process for the two types of erosion models (p < 0.001). Mg(OH)₂-containing dentifrices were effective in reducing enamel extrinsic acid erosion as determined by % SHL (p < 0.001) when compared to the control group, being better than positive control (p < 0.001); however, the dentifrices were not effective for the intrinsic model (p = 0.295). With regards to surface wear, no statistically significant differences were found among the groups for CA (p = 0.225) and HCl (p = 0.526).ConclusionThe findings suggest that Mg(OH)₂ dentifrices might protect enamel against slight erosion, but protection was not effective for stronger acid erosion.     

Appropriate real-time PCR reference genes for fluoride treatment studies performed in vitro or in vivo

ObjectiveQuantitative real-time PCR (qPCR) is routinely performed for experiments designed to identify the molecular mechanisms involved in the pathogenesis of dental fluorosis. Expression of reference gene(s) is expected to remain unchanged in fluoride-treated cells or in rodents relative to the corresponding untreated controls. The aim of this study was to select optimal reference genes for fluoride experiments performed in vitro and in vivo.DesignFive candidate genes were evaluated: B2m, Eef1a1, Gapdh, Hprt and Tbp. For in vitro experiments, LS8 cells derived from mouse enamel organ were treated with 0, 1, 3 and/or 5 mM sodium fluoride (NaF) for 6 or 18 h followed by RNA isolation. For in vivo experiments, six-week old rats were treated with 0 or 100 ppm fluoride as NaF for six weeks at which time RNA was isolated from enamel organs. RNA from cells and enamel organs were reverse-transcribed and stability of gene expression for the candidate reference genes was evaluated by qPCR in treated versus non-treated samples.ResultsThe most stably expressed genes in vitro according to geNorm were B2m and Tbp, and according to Normfinder were Hprt and Gapdh. The most stable genes in vivo were Eef1a1 and Gapdh. Expression of Ddit3, a gene previously shown to be induced by fluoride, was demonstrated to be accurately calculated only when using an optimal reference gene.ConclusionsThis study identifies suitable reference genes for relative quantification of gene expression by qPCR after fluoride treatment both in cultured cells and in the rodent enamel organ.     

Effect of biomimetic mineralization on enamel and dentin: A Raman and EDX analysis

ObjectiveTo investigate the effect of an experimental biomimetic mineralization kit (BIMIN) on the chemical composition and crystallinity of caries-free enamel and dentin samples in vitro.MethodsEnamel and dentin samples from 20 human teeth (10 for enamel; 10 for dentin) were divided into a control group without treatment and test samples with BIMIN treatment. Quantitative analysis of tissue penetration of fluoride, phosphate, and calcium was performed using energy-dispersive X-ray spectroscopy (EDX). Mineralization depth was measured by Raman spectroscopy probing the symmetric valence vibration near 960 cm⁻¹ as a marker for crystallinity. EDX data was statistically analyzed using a paired t-test and Raman data was analyzed using the Student’s t-test.ResultsEDX analysis demonstrated a penetration depth of fluoride of 4.10 ± 3.32 μm in enamel and 4.31 ± 2.67 μm in dentin. Calcium infiltrated into enamel 2.65 ± 0.64 μm and into dentin 5.58 ± 1.63 μm, while the penetration depths for phosphate were 4.83 ± 2.81 μm for enamel and 6.75 ± 3.25 μm for dentin. Further, up to 25 μm of a newly mineralized enamel-like layer was observed on the surface of the samples. Raman concentration curves demonstrated an increased degree of mineralization up to 5–10 μm into the dentin and enamel samples.SignificanceBiomimetic mineralization of enamel and dentin samples resulted in an increase of mineralization and a penetration of fluoride into enamel and dentin.     

Effect of full crown preparation on pulpal blood flow in man

ObjectiveTo determine if full crown preparation causes an increase in pulpal blood flow (PBF), indicating inflammation, in human subjects.DesignThe experiments were carried out on 35 intact, mandibular posterior teeth in 13 subjects: 32 were abutments for 16 fixed bridges that replaced first molars; the other 3 were first premolars adjacent to abutment teeth that served as un-operated controls. Crown preparations were made using an air-rotor with water-spray under regional block anaesthesia (4% articaine with epinephrine 1:100,000). PBF was recorded with a laser Doppler flow meter (LDF) before and after administering the anaesthetic, with the LDF probe on the buccal enamel. PBF was then recorded from the abutment teeth with the probe on buccal dentine after preparing the buccal surfaces of both teeth, after completing the crown preparations, and after 1 and 7 days. PBF was also recorded from the buccal enamel of the control teeth on each occasion.ResultsThe mean ± S.D. PBF values before and after anaesthesia were 2.63 ± 2.13 and 2.42 ± 2.38P.U. respectively, which were not significantly different (Paired t-test). The mean values for the abutment teeth after buccal preparation, after complete crown preparation, and after 1 and 7 days were 5.20 ± 2.49, 4.53 ± 2.52, 4.92 ± 2.98 and 5.48 ± 2.65P.U. respectively. The 4 values for each tooth were not significantly different (two-way RM ANOVA). In the control group, the values under all six conditions were not significantly different.ConclusionsRegional block anaesthesia produced no change in PBF, nor did full-crown preparation, neither immediately after the procedure nor 1 and 7 days later.     

奥乐V护牙剂对早期釉质龋再矿化作用的研究

目的 观察奥乐V护牙剂对早期釉质龋的再矿化作用。方法 将50个牛牙釉质块建立人工龋模型后随机分为5组：奥乐V护牙剂组、GC护牙素组、氟化钠（NaF）组、奥乐V护牙剂+NaF组、去离子水（DDW）组,分别进入 pH循环模型。采用显微硬度计测定各组标本脱矿前和pH循环后的釉质表面硬度值,扫描电镜观察各组再矿化后釉 质表面形态结构的改变。结果 各实验组经再矿化处理后其表面显微硬度均明显提高（P<0.001）,奥乐V护牙剂组再矿化后其表面显微硬度低于NaF组（P<0.001）,但与GC护牙素组间差异无统计学意义（P>0.05）;再矿化后奥乐V护牙剂组釉质表面有较大颗粒沉积。结论 奥乐V护牙剂具有促早期人工釉质龋再矿化的作用。