Transverse Microradiography Research Articles

Effectiveness of New Isomalt-Containing Toothpaste Formulations in Preventing Dental Caries: A Microbial Study.

Human dental enamel slabs were allocated to six groups (30/group) at random: De-ionized distilled water (DDW), and toothpaste containing 10% Isomalt, 1100 ppm fluoride, 0.05% cetylpyridinium chloride [CPC] (ICT); 10% Isomalt, 1100 ppm fluoride (IT); 10% Isomalt, 1100 ppm fluoride, 1.5% Sodium lauryl sulfate [SLS] (IST); 1100 ppm fluoride only (FT); 1100 ppm fluoride with SLS (FST). The enamel slabs were exposed to caries development via plaque growth in a Microbial Caries Model for 7 days. Toothpastes were applied as slurries (one toothpaste-three DDW) for 2 min twice daily. Demineralization was measured as the change in surface microhardness (ΔSMH) and amount of mineral lost (∆Z), and these metrics were assessed using Transverse Microradiography. Intra-group (SMH) and intergroup (%∆SMH and ∆Z) comparisons were paired t-test and Tukey's test (α = 0.05), respectively. With SMH, demineralization was found to be significant (p < 0.001) in all groups compared to sound enamel baseline, except ICT group. With %ΔSMH, all other groups had significantly (p < 0.001) less demineralization compared to DDW. Significantly (p < 0.001) greater demineralization was observed in IT, FT and FST compared to ICT, and no significant difference was observed between IST and ICT or FT. With ∆Z, relative to the DDW group, the inhibition of demineralization was significant (p < 0.0001) in all groups at varying percentages. Toothpaste containing 10% Isomalt, 1100 ppm fluoride, and 0.05% CPC demonstrated greater efficacy in inhibiting caries development amid dental plaque compared to toothpaste containing only 1100 ppm fluoride.

Lacticaseibacillus rhamnosus inhibits the development of dental caries in rat caries model and in vitro

ObjectivesDental caries result from a microbial imbalance in the oral cavity. Probiotics ecologically modulate the oral microflora to prevent caries. This study evaluated the anti-cariogenic effects of two Lacticaseibacillus rhamnosus strains in vitro and in vivo to provide a more theoretical basis for its clinical applications in caries prevention. MethodsIn the study, cariogenic biofilms were grown with L. rhamnosus (LGG) or L. rhamnosus ATCC 7469 and analyzed. Quantitative real-time PCR (qPCR), Scanning Electron Microscope (SEM), and Confocal laser scanning microscope (CLSM) were used to detect the changes in the composition and architectures; cariogenic activity was measured by the lactic acid production and Transverse Microradiography (TMR). The effects of LGG on the 12 Sprague-Dawley rat caries model were assessed using Keyes scores and micro-CT analysis. Oral microbiome changes were evaluated through 16S rRNA gene high-throughput sequencing. ResultsL. rhamnosus can reduce cariogenic bacteria in biofilm by 14.7 % to 48.9 %, with LGG exhibiting more potent inhibitory effects. Both strains of L. rhamnosus can adhere to the surface of biofilms, reduce the extracellular polysaccharides (EPS) matrix, and loosen the biofilm structure. L. rhamnosus inhibited cariogenic activity by reducing the lactic acid production in biofilms. The bovine enamel blocks presented lower mineral loss values and lesion depth values in the group Core+L.rh and Core+LGG. LGG-ingested rats had significantly lower levels of moderate dentin lesions and higher mineral density than the control group. The 16 s rRNA gene sequencing revealed that LGG regulated the beta diversity of the oral microbial community in the rat dental caries model. ConclusionsThis study revealed the promising potential of L. rhamnosus, especially the LGG strain, in the ecological prevention of dental caries. Clinical SignificanceProbiotics may provide a strategy for preventing caries by regulating the oral microecological balance. The study revealed the promising anti-caries potential of the LGG probiotic strain in vivo and in vitro. It is expected that LGG could be used as an oral probiotic for the clinical prevention and treatment of caries.

Influence of irradiated dentin, biofilm and different artificial saliva formulations on root dentin demineralization

The aim of this study was evaluated the influence of radiation as well as of new formulations of artificial saliva on the development of root dentin lesions. Bovine root samples were divided into: irradiated (70 Gy) dentin or not; the type of biofilm (from irradiated patient-experimental or non-irradiated patient-control) and the type of artificial saliva (for the condition irradiated dentin/biofilm from irradiated patient): Control Artificial Saliva (inorganic); Control Saliva + 1 mg/ml hemoglobin; Control Saliva +0.1 mg/ml cystatin; Control Saliva + hemoglobin + cystatin; Bioextra (positive control) and deionized water (DiW, negative control) (n = 12/group). Biofilm was produced using human biofilm and McBain saliva (0.2 % of sucrose, 37o C and 5 % CO2); the treatments were done 1x/day, for 5 days. Colony-forming units (CFU) counting was performed; demineralization was quantified by transversal microradiography. Two-way ANOVA/Bonferroni or Sidak test for the comparison between biofilm x dentin and ANOVA/Tukey or Kruskal-Wallis/Dunn for comparing artificial saliva were done (p < 0.05). The type of biofilm had no influence on CFU and demineralization. Sound dentin under control biofilm presented the lowest Lactobacillus ssp. and Streptococcus mutans CFU and the lowest mean mineral loss (R) (25.6 ± 2.2; 23.7 ± 2.9 %) compared to irradiated dentin (26.1 ± 2.8; 28.1 ± 3.3, p < 0.004) for both types of biofilms (experimental and control, respectively). Bioextra was the only artificial saliva that reduced R (10.8 ± 2.5 %) and Lesion Depth (LD) (35 ± 15 μm) compared to DiW (17.3 ± 3.3 %, 81 ± 18 μm, p < 0.0001). Irradiation has impact on caries development; the experimental saliva were unable to reduce its occurrence.

The damage and remineralization strategies of dental hard tissues following radiotherapy

ObjectivesThis study pursued two main purposes. The first aim was to expound on the microscopic factors of radiation-related caries (RRC). Further, it aimed to compare the remineralization effect of different remineralizing agents on demineralized teeth after radiotherapy.MethodsThe enamel and dentin samples of bovine teeth were irradiated with different doses of radiation. After analysis of scanning electron microscope (SEM), X-Ray diffraction (XRD), and energy dispersive spectrometer (EDS), the samples irradiated with 50 Gy radiation were selected and divided into the demineralization group, the double distilled water (DDW) group, the Sodium fluoride (NaF) group, the Casein phosphopeptide-amorphous calcium phosphate (CPP-ACP) group, the NaF + CPP-ACP group, and the Titanium tetrafluoride (TiF4) group. After demineralization, remineralizing agents treatment, and remineralization, the samples were evaluated using SEM, atomic force microscope (AFM), EDS, and transverse microradiography (TMR).ResultsA radiation dose of 30 Gy was sufficient to cause damage to the dentinal tubules, but 70 Gy radiation had little effect on the microstructure of enamel. Additionally, the NaF + CPP-ACP group and the TiF4 group significantly promoted deposit formation, decreased surface roughness, and reduced mineral loss and lesion depth of demineralized enamel and dentin samples after radiation.ConclusionsRadiation causes more significant damage to dentin compared to enamel. NaF + CPP-ACP and TiF4 had a promising ability to promote remineralization of irradiated dental hard tissues.Advances in knowledgeThis in vitro study contributes to determining a safer radiation dose range for teeth and identifying the most effective remineralization approach for RRC.

The effect of silver diammine fluoride on In Vitro Enamel caries lesion remineralization and staining as a function of lesion baseline mineral distribution

Objectivesto investigate whether baseline mineral distribution modulates the ability of silver diammine fluoride (SDF) to remineralize and stain enamel caries lesions. MethodsThis laboratory study followed a 3 [treatment: SDF/fluoride varnish (FV)/deionized water (DIW)] ×3 [lesion protocol: methylcellulose (MeC)/hydroxyethylcellulose (HEC)/Carbopol 907 (C907)] factorial design. Lesions were created in bovine enamel specimens (n = 20). Treatments were applied and lesions remineralized in artificial saliva. Digital transverse microradiography (TMR-D) was used to analyze lesions. Lesion color was monitored spectrophotometrically. The effects of lesion protocol and treatment on changes in lesion depth (ΔLD), mineral loss (ΔΔZ), maximum mineral density at the surface zone (ΔSZmax), and color changes related to remineralization (ΔL*remin) were analyzed using two-way ANOVA. ResultsThe treatment×lesion protocol interaction was significant for ΔΔZ (p < 0.01) and ΔL*remin (p < 0.01), however not for ΔLD (p = 0.23) or ΔSZmax (p = 0.91). There were no differences in ΔΔZ between treatments in HEC and C907 lesions. However, DIW resulted in more remineralization than both SDF (p < 0.01) and FV (p = 0.01) in MeC lesions. Considering changes from lesion baseline after remineralization in MeC lesions, SDF treatment resulted in the highest mineral gain in the surface zone. However, DIW revealed the highest mineral gain after remineralization in the lesion body. SDF stained lesions with the intensity increasing after remineralization in C907 lesions, whereas staining decreased in MeC and HEC lesions. ConclusionHigh fluoride treatments can interfere with continuous remineralization of caries lesions due to partial arrest. Baseline lesion mineral distribution affects SDF's ability to enhance remineralization and the staining caused by SDF. Clinical SignificanceSDF is being used to arrest active caries lesions extending into dentin and to treat dentin hypersensitivity. This study shed light on SDF's effect on an isolated process in dental caries only, remineralization. It achieved this by examining enamel caries lesions with differing mineral distributions and assessing their staining properties.

Enamel Caries Lesion Depth Obtained by Optical Coherence Tomography and Transverse Microradiography: A Comparative Study

Introduction: Visual imaging of subsurface caries lesions is of vital interest in dentistry, which can be obtained by invasive radiography technique as well as by available non-destructive imaging approaches. Thus, as a first step toward the development of a new innovative approach, Spectral-domain optical coherence tomography (SD-OCT) was applied to detect the lesion depth in comparison to the established reference technique (transverse microradiography [TMR]). Methods: Bovine enamel specimens were demineralized for 5 days, following previous studies. For OCT, the resulting artificial lesions were scanned three-dimensionally (SD-OCT) and semi-automated measured (CarLQuant). For TMR, specimens were sectioned and the lesion depth was manually determined (Inspektor Research System). Results: The range of lesion depth detected with OCT was 24.0–174.0 μm (mouth rinse study), 18.0–178.0 μm (toothpastes study) and with TMR 59.2–198.0 μm (mouth rinse study), 33.2–133.4 μm (toothpastes study). We found a strong correlation between both methods in terms of lesion depth (Spearman rankwith outlierp < 0.001, Rho = 0.75, Spearman rankwithout outlierp = 0.001, Rho = 0.79). The two methods produce similar results (Passing-Bablok regression, 1.16). As deeper is the lesion, the smallest is the difference between both methods as indicated by Bland-Altman-plots. Conclusion: Especially in the case of deep lesions, the values obtained by both methods are in agreement, and OCT can potentially substitute TMR to detect and assess lesion depth with the benefit of being non-destructive.

Effect of phosphate group on remineralization of early enamel caries regulated by amelogenin peptide.

To show the effect of the phosphate group on the remineralization process of early enamel caries mediated by amelogenin peptide. Freshly extracted, completed, and crack-free bovine teeth were used to create artificial early enamel caries, which were randomly divided into four groups: Group A: fluorination remineralized solution treatment group; Group B: pure remineralized solution treatment group. Group C: 100 g/ml recombinant Amelogenin peptide remineralized solution treatment group (with single phosphate group on N-terminus); Group D: 100 g/ml non-phosphorylated recombinant Amelogenin peptide remineralized solution treatment group (without single phosphate group on N-terminus). For 12 days, fresh remineralized solutions were replaced daily. Transverse microradiography (TMR) was used after remineralization to determine mineral loss and demineralization depth before and after each sample's remineralization. Each sample's depth of remineralization and mineral acquisition were then determined. The recombinant amelogenin peptide group significantly outperformed the non-phosphorylated amelogenin peptide group in terms of mineral acquisition and mineralization depth (P<0.05). The recombinant Amelogenin's solitary phosphate group at the N-terminus helps recombinant Amelogenin to encourage the remineralization process of early enamel caries.

Biomimetic Remineralization of Artificial Caries Lesions with a Calcium Coacervate, Its Components and Self-Assembling Peptide P11-4 In Vitro.

The application of calcium coacervates (CCs) may hold promise for dental hard tissue remineralization. The aim of this study was to evaluate the effect of the infiltration of artificial enamel lesions with a CC and its single components including polyacrylic acid (PAA) compared to that of the self-assembling peptide P11-4 in a pH-cycling (pHC) model. Enamel specimens were prepared from bovine incisors, partly varnished, and stored in demineralizing solution (DS; pH 4.95; 17 d) to create two enamel lesions per sample. The specimens were randomly allocated to six groups (n = 15). While one lesion per specimen served as the no-treatment control (NTC), another lesion (treatment, T) was etched (H3PO4, 5 s), air-dried and subsequently infiltrated for 10 min with either a CC (10 mg/mL PAA, 50 mM CaCl2 (Ca) and 1 M K2HPO4 (PO4)) (groups CC and CC + DS) or its components PAA, Ca or PO4. As a commercial control, the self-assembling peptide P11-4 (CurodontTM Repair, Credentis, Switzerland) was tested. The specimens were cut perpendicularly to the lesions, with half serving as the baseline (BL) while the other half was exposed to either a demineralization solution for 20 d (pH 4.95; group CC + DS) or pHC for 28 d (pH 4.95, 3 h; pH 7, 21 h; all five of the other groups). The difference in integrated mineral loss between the lesions at BL and after the DS or pHC, respectively, was analyzed using transversal microradiography (ΔΔZ = ΔZpHC - ΔZbaseline). Compared to the NTC, the mineral gain in the T group was significantly higher in the CC + DS, CC and PAA (p < 0.05, Wilcoxon). In all of the other groups, no significant differences between treated and untreated lesions were detected (p > 0.05). Infiltration with the CC and PAA resulted in a consistent mineral gain throughout the lesion body. The CC as well as its component PAA alone promoted the remineralization of artificial caries lesions in the tested pHC model. Infiltration with PAA further resulted in mineral gain in deeper areas of the lesion body.

Acquired enamel pellicle and biofilm engineering with a combination of acid-resistant proteins (CaneCPI-5, StN15, and Hemoglobin) for enhanced protection against dental caries - in vivo and in vitro investigations.

This study was designed in two-legs. In the in vivo, we explored the potential of a rinse solution containing a combination (Comb) of 0.1mg/mL CaneCPI-5 (sugarcane-derive cystatin), 1.88 × 10- 5M StN15 (statherin-derived peptide) and 1.0mg/mL hemoglobin (Hb) to change the protein profile of the acquired enamel pellicle(AEP) and the microbiome of the enamel biofilm. The in vitro, was designed to reveal the effects of Comb on the viability and bacterial composition of the microcosm biofilm, as well as on enamel demineralization. In vivo study, 10 participants rinsed (10mL,1min) with either deionized water (H2O-control) or Comb. AEP and biofilm were collected after 2 and 3h, respectively, after rinsing. AEP samples underwent proteomics analysis, while biofilm microbiome was assessed via 16S-rRNA Next Generation Sequencing(NGS). In vitro study, a microcosm biofilm protocol was employed. Ninety-six enamel specimens were treated with: 1)Phosphate-Buffered Solution-PBS(negative-control), 2)0.12%Chlorhexidine, 3)500ppmNaF and 4)Comb. Resazurin, colony-forming-units(CFU) and Transversal Microradiography(TMR) were performed. The proteomic results revealed higher quantity of proteins in the Comb compared to control associated with immune system response and oral microbial adhesion. Microbiome showed a significant increase in bacteria linked to a healthy microbiota, in the Comb group. In the in vitro study, Comb group was only efficient in reducing mineral-loss and lesion-depth compared to the PBS. The AEP modification altered the subsequent layers, affecting the initial process of bacterial adhesion of pathogenic and commensal bacteria, as well as enamel demineralization. Comb group shows promise in shaping oral health by potentially introducing innovative approaches to prevent enamel demineralization and deter tooth decay.

Acquired Pellicle and Biofilm Engineering by Rinsing with Hemoglobin Solution

Introduction: The identification of acid-resistant proteins, including hemoglobin (Hb), within the acquired enamel pellicle (AEP) led to the proposition of the “acquired pellicle engineering” concept, which involves the modification of the AEP by incorporating specific proteins, presenting a novel strategy to prevent dental demineralization. Objective: Combining in vivo and in vitro proof-of-concept protocols, we sought to reveal the impact of AEP engineering with Hb protein on the biofilm microbiome and enamel demineralization. Methods: In the in vivo studies, 10 volunteers, in 2 independent experiments, rinsed (10 mL,1 min) with deionized water-negative control or 1.0 mg/mL Hb. The AEP and biofilm formed along 2 or 3 h, respectively, were collected. AEP was analyzed by quantitative shotgun-label-free proteomics and biofilm by 16S-rRNA next-generation sequencing (NGS). In in vitro study, a microcosm biofilm protocol was employed. Seventy-two bovine enamel specimens were treated with (1) phosphate-buffered solution (PBS), (2) 0.12% chlorhexidine, (3) 500 ppm NaF, (4) 1.0 mg/mL Hb, (5) 2.0 mg/mL Hb, and (6) 4.0 mg/mL Hb. The biofilm was cultivated for 5 days. Resazurin, colony forming units (CFU), and transversal microradiography were performed. Results: Proteomics and NGS analysis revealed that Hb increased proteins with antioxidant, antimicrobial, acid-resistance, hydroxyapatite-affinity, calcium-binding properties and showed a reduction in oral pathogenic bacteria. In vitro experiments demonstrated that the lowest Hb concentration was the most effective in reducing bacterial activity, CFU, and enamel demineralization compared to PBS. Conclusion: These findings suggest that Hb could be incorporated into anticaries dental products to modify the oral microbiome and control caries, highlighting its potential for AEP and biofilm microbiome engineering.

Remineralisation of enamel and dentine with stabilised stannous fluoride dentifrices in a randomised cross-over in situ trial

ObjectivesTo compare the remineralisation efficacy and ion bioavailability of two novel SnF2-containing dentifrices in a blinded, cross-over, randomised in situ clinical study. MethodsSix participants wore removal palatal appliances holding human enamel and dentine blocks with subsurface lesions. Appliances were worn for two treatment periods of 14 consecutive days each, with a one-week washout period in-between. Participants were randomly allocated to rinse with a 1:5 diluted coded slurry of one of two dentifrices containing either 5 % casein phosphopeptide-amorphous calcium phosphate (CPP-ACP) +1100 ppm F as SnF2 [MIPOP], or 1100 ppm F as SnF2 [CT], for 1 min, four times a day. Saliva was collected post-treatment and analysed for tin, calcium, inorganic phosphate and fluoride ions using atomic absorption spectrophotometry and ion chromatography. Enamel and dentine lesions were analysed for percent remineralisation (%R) using transverse microradiography and percent surface microhardness recovery (%SMHR). ResultsMIPOP released significantly higher F (3.00 ± 0.27 mM), Ca (15.23 ± 3.23 mM) and Sn (1.18 ± 0.13 mM) into saliva whereas CT released 2.89 ± 0.32 mM F and only 0.84 ± 0.11 mM Ca and 0.28 ± 0.10 mM Sn. MIPOP produced significantly higher %R than CT: 25.6 ± 1.5 % compared to 15.2 ± 0.7 % in enamel, and 33.6 ± 3.1 % compared to 20.6 ± 1.1 % in dentine. Additionally, MIPOP produced significantly higher %SMHR (18.2 ± 7.9 %) compared to CT (4.1 ± 0.6 %). ConclusionsBoth dentifrices promoted remineralisation, but the MIPOP dentifrice with added CPP-ACP and the ion-stabilising effects of CPP released higher amounts of bioavailable tin and produced significantly higher remineralisation and surface microhardness recovery. Clinical SignificanceModern dentifrices contain SnF2 for a range of oral health benefits. Challenges associated with stability of these formulations can affect ion bioavailability, reducing efficacy. Two dentifrices with SnF2 promoted remineralisation in situ, however the dentifrice with the added saliva biomimetic CPP-ACP was superior and therefore may produce greater health benefits.

Effect of calcium-coacervate infiltration of artificial enamel caries lesions in de- and remineralizing conditions

ObjectivesCalcium-coacervate emulsions (CC) might be considered as mineral precursors to foster remineralization of carious dental hard tissues. This study analyzed the instant effect of repeated infiltration of artificial caries lesions with a CC emulsion as well as the effects of subsequent exposure of CC-infiltrated lesions to demineralizing and remineralizing environments. MethodsBovine enamel specimens were partly covered with varnish to leave three exposed windows. Artificial enamel caries lesions were created (pH 4.95, 17d). Baseline controls (BL) were obtained by preparing a thin section of each specimen. Specimens were allocated to five groups. In three groups lesions were etched with 37 % phosphoric acid gel, infiltrated with dipotassium hydrogen phosphate and subsequently with a calcium coacervate emulsion, prepared by mixing CaCl2 ⋅ 2H2O with polyacrylic acid sodium salt (PAA-Na). Subsequently, the infiltration effect was either analyzed immediately (Inf.) or after exposition to either de- (Inf.+DS) or remineralizing solution (Inf.+RS) for 10 or 20 days, respectively. In two control groups specimens were exposed to either DS or RS, respectively without prior CC infiltration. Integrated mineral loss [ΔZ(vol%×µm)] was analyzed using transverse microradiography (TMR). ResultsInfiltration of enamel caries lesions with coacervate solution resulted in only subtle immediate mineral gain even if repeated. When exposed to demineralizing conditions, infiltrated lesions showed significantly less mineral loss compared to untreated controls (p < 0.05; Kruskal Wallis) and exhibited characteristic mineral depositions within the lesion body. ConclusionsWhile immediate mineral gain by infiltration was only modest, the CC-emulsion might be able to prevent demineralization in acidic conditions. Clinical significanceCalcium coacervates might act protective against further demineralization when infiltrated into enamel caries lesions.

Fluoride-resistant Streptococcus mutans within cross-kingdom biofilms support Candida albicans growth under fluoride and attenuate the in vitro anti-caries effect of fluorine.

Fluoride-resistant Streptococcus mutans (S. mutans) might affect the ecological balance of biofilms in the presence of fluoride. We used a S. mutans and Candida albicans (C. albicans) cross-kingdom biofilm model to investigate whether fluoride-resistant S. mutans in biofilms would support C. albicans growth under fluoride stress and attenuate the in vitro anti-caries effect of fluorine. The impact of fluoride-resistant S. mutans on formation of cross-kingdom biofilms by S. mutans and C. albicans in the presence of fluoride was investigated in vitro using the crystal violet staining assay. Biofilm constitution was determined using colony-forming unit (CFU) counts and fluorescent in situ hybridization (FISH). Extracellular polysaccharide (EPS) generation in biofilms was determined by EPS/bacterial dying and water-insoluble polysaccharide detection. Acid production and demineralization were monitored using pH, lactic acid content, and transversal microradiography (TMR). The gene expression of microorganisms in the cross-kingdom biofilm was measured using qRT-PCR. Our results showed that both C. albicans and fluoride-resistant S. mutans grew vigorously, forming robust cross-kingdom biofilms, even in the presence of sodium fluoride (NaF). Moreover, fluoride-resistant S. mutans-containing cross-kingdom biofilms had considerable cariogenic potential for EPS synthesis, acid production, and demineralization ability in the presence of NaF than fluoride-sensitive S. mutans-containing biofilms. Furthermore, the gene expression of microorganisms in the two cross-kingdom biofilms changed dissimilarly in the presence of NaF. In summary, fluoride-resistant S. mutans in cross-kingdom biofilms supported C. albicans growth under fluoride and might attenuate the anti-caries potential of fluorine by maintaining robust cross-kingdom biofilm formation and cariogenic virulence expression in vitro in the presence of NaF.

In vitro effect of TiF4/NaF solution on the development of radiation-induced dentin caries.

To evaluate the protective effect of an experimental solution containing TiF4/NaF on the development of radiation-induced dentin caries lesions. bovine root samples were irradiated (70Gy) and distributed as following (n=12/group): Commercial Saliva (BioXtra), NaF (500 ppm F-), TiF4 (500 ppm F), TiF4/NaF (TiF4: 300 ppm F-, NaF: 190 ppm F-), and Phosphate buffer solution (PBS, negative control). Biofilm was produced using biofilm from irradiated patients and McBain saliva (0.2% of sucrose, at 37oC and 5% CO2) for five days. The treatments were applied 1x/day. Colony-forming units (CFU) were counted and demineralization was quantified by transversal microradiography. The ANOVA/Tukey test was applied for all parameters. All treatments reduced CFU for total microorganisms. TiF4 reduced Lactobacillus sp. (7.04±0.26 log10 CFU/mL) and mutans streptococci (7.18±0.28) CFU the most, when compared to PBS (7.58±0.21 and 7.75±0.17) and followed by NaF (7.12±0.31 and 7.34±0.22) and TiF4/NaF (7.16±0.35 and 7.29± 0.29). TiF4 and Commercial saliva showed the lowest integrated mineral loss (ΔZ-vol%.mm) (1977±150 and 2062±243, respectively) when compared to PBS (4540±335), followed by NaF (2403±235) and TiF4/NaF (2340±200). Commercial saliva was the only to significantly reduce mineral loss (LD-µm) (111±25) compared to PBS (153±24).Mean mineral loss (R-vol%) decreased by 35.2% for TiF4 (18.2±3.3) when compared to PBS (28.1±2.9) Conclusion: TiF4/NaF has a comparable anti-cariogenic effect to TiF4 and Commercial saliva under the model in this study.

Encapsulation of a novel peptide derived from histatin-1 in liposomes against initial enamel caries in vitro and in vivo.

Biomimetic mineralization mediated by proteins and peptides is a promising strategy for enamel repair, and its specific application model needs more research. In this work, we exploited a liposomal delivery system for a novel peptide (DK5) derived from histatin-1 (DK5-Lips) as a new biomimetic mineralization strategy against initial enamel caries. The DK5-Lips was prepared using calcium acetate gradient method and then the in vitro release, salivary stability, and cytotoxicity were studied. Initial enamel caries was created in bovine enamel blocks and subjected to pH-cycling model treated with DK5-Lips. Surface microhardness testing, polarized light microscopy (PLM), and transverse microradiography (TMR) were analyzed. Then the biocompatibility of DK5-Lips was evaluated in the caries model of Sprague-Dawley rats, and the anti-caries effect was assessed using Micro-CT analysis, Keyes scores, and PLM in vivo. DK5-Lips provided a mean particle size of (97.63 ± 4.94)nm and encapsulation efficiency of (61.46 ± 1.44)%, exhibiting a sustained release profile, excellent stability in saliva, and no significant toxicity on human gingival fibroblasts (HGFs). The DK5-Lips group had higher surface microhardness recovery, shallower caries depth, and less mineral loss in bovine enamel. Animal experiments showed higher volume and density values of residual molar enamel, lower Keyes score, and shallower lesion depth of the DK5-Lips group with good biocompatibility. As a safe and effective application model, DK5-Lips could significantly promote the remineralization of initial enamel caries both in vitro and in vivo. The potential of liposome utilization as vehicle for oral delivery of functional peptides may provide a new way for enamel restoration.

Xylitol associated or not with fluoride: Is the action the same on de- and remineralization?

ObjectivesThis study evaluated the effect of xylitol combined or not with fluoride (F) on reduction of demineralization and increase of remineralization of shallow and deep artificial enamel lesions. MethodsBovine enamel samples were allocated to the following solutions groups: no xylitol (negative control), 5% xylitol, 10% xylitol, 20% xylitol, 500 ppm F (as NaF), 5% xylitol+F, 10% xylitol+F or 20% xylitol+F (n = 12–15). For the demin study, a pH-cycling model (demineralization-6 h, pH 4.7/remineralization 18 h, pH 7.0) was employed for 7 days. Treatments were applied 2 × 1 min. In the remin study, specimens were pre-demineralized for 2, 5 or 10 days. Afterwards, a pH-cycling protocol was conducted (2 h demineralizing and 22 h remineralizing solution/day for 8 days) and the same treatments were done. The response variables were percentage surface hardness loss (%SHL) and transverse microradiography. Data were analyzed by RM ANOVA/Tukey or Kruskal-Wallis/Dunn (p < 0.05) ResultsF and Xylitol combined with F reduced the %SHL (23–30%) compared to the negative control (61.5%). The integrated mineral loss and the lesion depth were not reduced by any treatment. Surface hardness recovery was seen only for shallow lesions in case of 20% xylitol+F compared to negative control. No lesion depth recovery, but significant mineral recovery was seen for F (2-days and 10-days lesion). ConclusionsAll concentrations of xylitol+F reduced enamel surface demineralization, while only 20% xylitol+F improved surface remineralization of shallow lesions in vitro. Clinical significanceOur results suggest that while F or any concentration of xylitol + F reduces surface demineralization, only 20% xylitol+F improves surface remineralization of shallow lesions in vitro. Therefore, xylitol may be added into oral products, combined to F, to control dental caries.

Effect of Three Chemical Agents on Stain Removal from Dentin Caries Lesions Treated with Silver Diamine Fluoride

Introduction: This in vitro study aimed to evaluate the effect of three different chemical agents on stain removal and mineral uptake of artificial dentin caries (ADC) lesions treated with silver diamine fluoride (SDF). Methods: Baseline L*a*b* values were determined in polished human permanent dentin blocks, and ADC lesions were induced with an acid gel for 1 week. Samples were assigned to four groups; in three groups, half of each sample received SDF (30% SDF for 3 min), while the other half received SDF followed by a bleaching treatment protocol (garlic extract, bentonite, or 35% hydrogen peroxide). The fourth group had one SDF-treated half and one half without SDF. Color changes (ΔE) were measured by spectrometry, and transversal microradiography was used to quantify integrated mineral loss (ΔZ) 24 h after treatment (SDF or SDF + bleaching). A two-way mixed ANOVA was applied to thirty percent. Results: SDF application increased mineral uptake by ADC (p = 0.001). The type of chemical agent evaluated (p < 0.0001), time (p = 0.01), and their interaction (p < 0.0001) bleached the ADC treated with SDF. However, 35% hydrogen peroxide was the only compound with a bleaching effect (p < 0.001), without returning to baseline color. None of the compounds altered the mineral uptake effect of SDF (p = 0.30). Conclusion: This in vitro study showed mineral uptake effect in ACD within 24 h after SDF application and the ability of hydrogen peroxide to partially remove (reduction of 24%) the staining caused by SDF without affecting its mineral uptake effect.

Evaluation of Iontophoresis as a Tool in Comparison to Topical Remineralization Systems by Transverse Microradiography and Polarized Light Microscopy: An In Vitro Study.

To compare the efficacy of casein phosphopeptide-amorphous calcium phosphate (CPP-ACP) (GC Tooth Mousse) and tricalcium phosphate formulation (Clinpro) by topical application and iontophoresis. Noninvasive treatment of dental caries is a major advance in the clinical management of the disease. A total of 20 primary anterior and 20 primary molars were included in the study to be analyzed by transverse microradiography (TMR) and polarized light microscopy (PLM). The samples were coated with acid-resistant varnish, leaving a window on the buccal/lingual surface of enamel and immersed in demineralizing solution for 96 hours. Each sample was coated with varnish on the left half (control), and the right half served as a test. All the samples were divided into four groups. The test windows of specimens in groups I (GC Tooth Mousse) and II (Clinpro) were subjected to remineralization treatments for 10 days. Similarly, groups III and IV were subjected to iontophoresis using GC Tooth Mousse and Clinpro for 7 minutes. The samples were analyzed by TMR and PLM. There was no significant difference in the mean difference of mineral loss among the four groups. There was a significant difference in the mean difference of lesion depth among the four groups. Both iontophoresis and topical application were equally efficient, but one remineralization by iontophoresis equals 10 topical applications. VC R, Muppa R, Nallanchakrava S, et al. Evaluation of Iontophoresis as a Tool in Comparison to Topical Remineralization Systems by Transverse Microradiography and Polarized Light Microscopy: An In Vitro Study. Int J Clin Pediatr Dent 2023;16(S-1):S85-S90.

Using Antimicrobial Photodynamic Therapy with Ultrasound Devices and Bioactive Glasses as a Combined Approach for Treating Dentin Caries Lesions.

Novel approaches for caries lesion removal and treatment have been proposed. This study evaluates the combined use of an experimental ultrasound, aPDT (antimicrobial photodynamic therapy) and bioactive glasses on the removal, decontamination and remineralization of dentin caries lesions. A biological model created with a duo species biofilm (Streptococcus mutans and Lactobacillus acidophilus) was used for the development of a caries-like lesion over the dentin for 7 days. Bovine dentin specimens (4 × 4 × 2 mm) were randomized according to the following caries removal techniques: bur (BUR) or ultrasound (ULT), decontamination (with or without aPDT) and remineralization materials (45S5 or F18 bioactive glasses). The following different groups were investigated: caries lesion (control); sound dentin (control); BUR; BUR + aPDT; ULT; ULT + aPDT; BUR + 45S5, BUR + F18; ULT + 45S5; ULT + F18; BUR + aPDT + 45S5; BUR + aPDT + F18; ULT + aPDT + 45S5; and ULT + aPDT + F18. Transverse microradiography (TMR), cross-sectional microhardness (CSH), FT-Raman spectroscopy and confocal microscopy (CLSM) were performed. A two-way ANOVA and Tukey's test were used (α = 0.05). (3) Results: The TMR revealed a lesion depth of 213.9 ± 49.5 μm and a mineral loss of 4929.3% vol.μm. The CSH increases as a function of depth, regardless of the group (p < 0.05). Removal with BUR (24.40-63.03 KHN) has a greater CSH than ULT (20.01-47.53 KHN; p < 0.05). aPDT did not affect the CSH (p > 0.05). No difference was observed between 45S5 or F18 (p > 0.05), but a change was observed for ULT (p > 0.05). The FT-Raman shows no differences for the phosphate (p > 0.05), but a difference is observed for the carbonate and C-H bonds. The CLSM images show that aPDT effectively inactivates residual bacteria. A combination of ULT, aPDT and bioactive glasses can be a promising minimally invasive treatment.

Quantitative analysis of the degree of demineralization for bleached enamel by optical coherence tomography

BackgroundTooth bleaching imparts whitening effects along with adverse effects such as increased tooth sensitivity and enamel surface changes. Herein, we employed optical coherence tomography (OCT), a nondestructive optical detection technique, for evaluation of tooth enamel after treatment with peroxide-based bleaching agents. MethodsFifteen enamel samples were bleached using 38% acidic hydrogen peroxide-based bleach, subjected to OCT scanning, and then cross-sectioned and imaged under polarized light microscopy (PLM) and transverse microradiography (TMR). OCT cross-sectional images were compared with PLM and TMR. The depth and severity of demineralization produced in the bleached enamel were measured by OCT, PLM, and TMR. Comparison between the three techniques was performed using Kruskal-Wallis H non-parametric test and Pearson correlation. ResultsIn comparison with PLM and TMR, OCT clearly detected the changes in the enamel surface after hydrogen peroxide bleaching. Significant correlations (p<0.05) were observed in lesion depth between OCT and PLM (r=0.820), OCT and TMR (r=0.822), and TMR and PLM (r=0.861). There was no statistically significant difference in demineralization depth values measured by OCT, PLM, and TMR (p>0.05). ConclusionOCT can allow real-time, non-invasive imaging of artificially bleached tooth models and automatically measure the early changes in the enamel lesion structure upon exposure to hydrogen peroxide-based bleaching agents.