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.

Caries status in 12-year-old children, geographical location and socioeconomic conditions across European countries: A systematic review and meta-analysis.

BACKGROUND: Understanding of socioeconomic context might enable more efficient evidence-based preventive strategies in oral health. AIM: The study assessed the caries-related socioeconomic macro-factors in 12-year-olds across European countries. DESIGN: This systematic review involved epidemiological surveys on the caries status of 12-year-olds from 2011 to 2022. DMFT was analyzed in relation to gross national income (GNI), United Nations Statistical Division geographical categorization of European countries (M49), unemployment rate, Human Development Index (HDI), and per capita expenditure on dental health care. A meta-analysis was performed for countries reporting data on DMFT, stratified by GNI, and geographical location of European countries, using a random-effects model. RESULTS: The study involved 493 360 children from 36 countries in the geographic region of Europe. The analysis confirmed a strong negative correlation between income and caries experience (p < .01). Children living in higher-income countries showed 90% lower odds of poor oral health than in middle-income countries. Children living in West Europe showed 90% lower odds of poor oral health than children living in East Europe. CONCLUSION: The strong effect of macro-level socioeconomic contexts on children's oral health suggests favoring upstream preventive oral health strategies in countries with economic growth difficulties, Eastern and Southern parts of Europe.

Distribution of initial caries lesions in relation to fixed orthodontic therapy. A systematic review and meta-analysis.

BACKGROUND: Initial caries lesion (ICLs) adjacent to orthodontic brackets are the most common side effect of orthodontic treatment with fixed appliances. The reported prevalence is uncertain and varies considerably across studies, from 27% to 97%. OBJECTIVES: This paper was designed to evaluate and synthesize the available evidence on the prevalence and incidence rates of ICLs in relation to orthodontic treatment. Selection criteria: The review (Prospero protocol CRD42023412952) included randomized and non-randomized clinical trials of interventions, cohort studies, and cross-sectional studies, published after 1990 on the prevalence or incidence of ICLs during or after orthodontic treatment with fixed appliances. Search methods: Pubmed, Scopus, and Embase databases were searched from 1990 until 01 May 2023. The risk of bias assessment was performed with RoB 2 and ROBINS-I tool and the Joanna Briggs Institute Critical Appraisal Checklist. Data collection and analysis: The proportion of individuals with ICLs, reported as the number/percentage of individuals/teeth with ICLs or mean number of ICLs per subject, were used to synthesize results. RESULTS: The search yielded a total of 468 papers; 21 studies were included in the systematic review, 2 of which were not included in the meta-analysis. The prevalence rate [95%CI] of ICLs was 0.57% [0.48; 0.65] in 1448 patients, 0.22% [0.14; 0.33] in 11583 teeth, with a mean number of lesions equal to 2.24 [1.79; 2.70] in 484 patients evaluated. The incidence rate of new carious lesions developed during orthodontic treatment was 0.48% [0.33; 0.63] in 533 patients, 0.15% [0.08; 0.26] in 1890 teeth with a mean number of ICLs equal to 2.29 [1.12; 3.46] in 208 patients evaluated. LIMITATIONS: Although the high number of included studies and the overall good quality, there was a significant heterogeneity in the collected data. CONCLUSION: The prevalence and incidence rates of ICLs in subjects undergoing orthodontic treatment are quite high and raise some concerns in terms of risk assessment of orthodontic treatment. ICLs represent an alarming challenge for both patients and professionals. Effective caries prevention strategies during treatment need to be considered and implemented where appropriate.

Design, development and validation of a questionnaire to assess dentists' knowledge and experience in diagnosing, recording, and managing root caries.

OBJECTIVES: The prevalence of root caries is increasing globally, especially in the elderly population, and even though the number of patients with root caries lesions is augmenting, there are still many discrepancies in how dentists manage this condition. The present study aimed to develop and validate a questionnaire to evaluate how dentists diagnose, record and manage root caries lesions, and to verify the validity and reliability of this questionnaire. MATERIALS AND METHODS: An expert panel developed a self-administered questionnaire survey with three domains: (1) dentists' knowledge on diagnosis, recording, and managing root caries; (2) information about their current general clinical routines; (3) their demographics. The original English [E] version was translated into three different languages (French [F], German [G], Italian [I]), and subsequently back-translated into English by independent dentists. For the validation, 82 dentists (20-22 for each of the translated versions) accepted to answer the questionnaire at two different time-points (with 1-week interval). The data was quality checked. Construct validity, internal reliability, and intra-class correlation (ICC) were assessed. RESULTS: Seventy-seven dentists completed the questionnaire twice [E: 17; F: 19; G: 19; I: 22]. The mean ICC (standard deviation) was 0.98(0.03) for E, 0.90(0.12) for F, 0.98 (0.04) for G, and 0.98 (0.01) for I. Overall, the test-retest reliability was excellent (mean ICC (SD): 0.96 (0.08)). Furthermore, the questionnaire demonstrated good internal reliability (inter-observer reliability; Fleiss kappa: overall:0.27(fair); E:0.30 (fair); F: 0.33(fair); G: 0.33(fair); I: 0.89 (almost perfect)). CONCLUSION: The questionnaire was validated and is suitable to be used in the four languages to assess the knowledge of dentists on diagnosing, recording and managing root caries. CLINICAL SIGNIFICANCE: The present questionnaire was validated and seems to be a good tool to evaluate how dentists diagnose, record, and manage root caries lesions both in its original (English) and its translated (French, German, and Italian) versions.

Correlation of quantitative light-induced fluorescence and qualitative visual rating in infiltrated post-orthodontic white spot lesions.

OBJECTIVES: The aim of this prospective, clinical single-centre study was to evaluate the masking efficacy of post-orthodontic resin infiltration after 12-month follow-up and correlate quantitative and qualitative outcome measures. METHODS: Patients with completed fixed orthodontic treatment and the presence of one or more vestibular active non-cavitated white spot lesion/s (WSL) [ICDAS 1 or 2 (International Caries Detection and Assessment System)] were provided with resin infiltration 3-12 months after bracket removal. All patients (n = 31) participating before (t0) intervention were invited again and examined after 12 months (t2). Enamel demineralization was scored using quantitative light-induced fluorescence [QLF (DeltaF[flourescence], DeltaQ[lesion volume], White Spot Area)] and qualitative visual rating [11-point Likert-scale from 0 (no lesions visible on any tooth) to 10 (all teeth affected on the entire vestibular surface)]. RESULTS: In 17 patients (7 female and 10 male) 112 WSL (ICDAS 1: n = 1; ICDAS 2: n = 111) in 112 teeth were (re)examined. Before treatment (t0) a significant, weak (DeltaF), and moderate (DeltaQ, White Spot Area) correlation was observed between the quantitative and the qualitative rating (P < 0.002) [median DeltaF: -7.31 (-10.4/-6.58)%; DeltaQ:-2.25 (-10.8/-0.41)% mm2; White Spot Area: 0.34 (0.05/1.16) mm2; visual rating:3.7 ± 1.2]. Resin infiltration led to significantly increased fluorescence and decreased visual scores (P < 0.001) 7 days (t1) and 12 months (t2) after treatment. No significant changes based on DeltaF [-6.55 (-7.29/-6.08)%] and on visual ratings [1.0 ± 1.0] were observed between t1 and t2 (P = 1.000). After 7 days (t1) the correlation between the quantitative and the qualitative ratings remained significant, weak to moderate (P < 0.002). After 12 months (t2) the correlation was (non-)significant and weak for DeltaF, DeltaQ, and White Spot Area (P ≤ 0.097). LIMITATIONS: Since the overall masking efficacy of resin infiltration has been shown previously, an untreated control group was omitted. CONCLUSIONS: When assessing the masking efficacy of infiltrated post-orthodontic WSL only a weak to moderate correlation was found between QLF values and visual ratings. Furthermore, over time this correlation decreased. Thus, it remains unclear if QLF is a viable method to assess and quantify infiltrated post-orthodontic WSL over time. TRIAL REGISTRATION: German Clinical Trials Register (DRKS-ID:DRKS00005067).

Effects of the association of high fluoride- and calcium-containing caries-preventive agents with regular or high fluoride toothpaste on enamel: an in vitro study.

OBJECTIVES: The aim of this in vitro study was to compare the caries-preventive effect of various high fluoride- and calcium-containing caries-preventive agents (> 22.000 ppm F- [ppm]) in adjunct to use of regular (1450 ppm) or high (5000 ppm) fluoride toothpaste on sound as well as demineralized enamel. MATERIALS AND METHODS: Bovine enamel specimens (n = 276; 5 mm × 3.5 mm × 3 mm) having one sound surface [ST] and one artificial caries lesion [DT] were randomly allocated to 12 groups. Interventions before pH-cycling were no intervention ([SC1/SC5]), application of varnishes/solutions containing NaF (22,600 ppm; Duraphat [NaF1/NaF5]); NaF + tricalcium phosphate (22,600 ppm; Clinpro White Varnish [TCP1/TCP5]); NaF + CPP-ACP (22,600 ppm; MI Varnish [CPP1/CPP5]); silver diammine fluoride (35,400 ppm; Cariestop 30%[SDF1/SDF5]); and NaF + calcium fluoride (45,200 ppm; Biophat[CaF1/CaF5]). During pH-cycling (28 days, 6 × 120 min demineralization/day) half of the specimens in each group were brushed (10 s; 2 × /day) with either 1,450 (NaF; named, e.g., TCP1) or 5,000 ppm (NaF; e.g., TCP5) dentifrice slurry. Differences in integrated mineral loss (∆∆Z) and lesion depth (∆LD) were calculated between values after initial demineralization and after pH-cycling using transversal microradiography. RESULTS: After pH-cycling, SC1/SC5 showed significantly increased ∆ZDT/LDDT values, indicating further demineralization (p < 0.05; paired t-test). Decreased ∆ZDT values, indicating non-significant remineralization, could only be observed in CaF1/CaF5 (p > 0.05; paired t-test). Additional use of all varnishes/solutions significantly decreased ∆∆ZDT/∆∆ZST and ∆LDDT/∆LDST compared to SC1/SC5 (p < 0.05;ANCOVA). Between 1450 and 5000 ppm dentifrices, a significant difference in ∆∆ZDT/∆∆ZST and ∆LDDT/∆LDST could only be observed for SC1/SC5 (p < 0.05; ANCOVA). CONCLUSION: Under the conditions chosen, all fluoride varnishes/solutions significantly reduced demineralization. Furthermore, a significant dose-response characteristic for fluoride varnishes could be revealed. However, no additional benefit could be observed, when varnishes were combined with high fluoride instead of regular fluoride dentifrices. CLINICAL RELEVANCE: For children and adolescents with high caries risks varnishes containing more than 22,600 ppm should be further investigated, as they offered higher caries-preventive effects in vitro. Furthermore, there seems to be no difference in the demineralization-inhibitory capacity of fluoride varnishes when used in combination with either standard or highly fluoridated dentifrices.

Increasing dental zirconia micro-retentive aspect through ultra-short pulsed laser microstructuring: study on flexural strength and crystal phase characterization.

OBJECTIVES: Although ultra-short pulsed laser (USPL) microstructuring has previously improved zirconia bond-strength, it is yet unclear how different laser-machined surface microstructures and patterns may influence the material's mechanical properties. Therefore, the aim of this study was to assess the flexural strength of zirconia after different USPL settings creating three different geometrical patterns with structures in micrometer scale. METHODS: One hundred sixty zirconia bars (3Y-TZP, 21 × 4 × 2.1 mm) were prepared and randomly divided into five groups (n = 32): no surface treatment (negative control-NC); sandblasting with Al2O3 (SB); and three laser groups irradiated with USPL (Nd:YVO4/1064 nm/2-34 J/cm2/12 ps): crossed-lines (LC), random-hatching (LR), and parallel-waves (LW). Bars were subjected to a four-point flexural test (1 mm/min) and crystal phase content changes were identified by X-ray diffraction. Surface roughness and topography were analyzed through 3D-laser-profilometry and SEM. Data were analyzed with parametric tests for roughness and Weibull for flexural strength (α = 5%). RESULTS: LR (Mean[95%CI]: 852.0 MPa, [809.2-894.7]) was the only group that did not show a significantly different flexural strength than NC (819.8 MPa, [796.6-842.9]), (p > 0.05). All laser groups exhibited higher Weibull moduli than NC and SB, indicating higher reliability and homogeneity of the strength data. An increase of monoclinic phase peak was only observed for SB. CONCLUSION: In conclusion, USPL created predictable, homogeneous, highly reproducible, and accurate surface microstructures on zirconia ceramic. The laser-settings of random-hatching (12 ps pulses) increased 3Y-TZP average surface roughness similarly to SB, while not causing deleterious crystal phase transformation or loss of flexural strength of the material. Furthermore, it has increased the Weibull modulus and consequently material's reliability. CLINICAL SIGNIFICANCE: Picosecond laser microstructuring (LR conditions) of 3Y-TZP ceramic does not decrease its flexural strength, while increasing materials realiability and creating highly reproducible and accurate microstructures. These features may be of interest both for improving clinical survival of zirconia restorations as well as enhancing longevity of zirconia implants.

Demineralization Inhibitory Effects of Highly Concentrated Fluoride Dentifrice and Fluoride Gels/Solutions on Sound Dentin and Artificial Dentin Caries Lesions in vitro.

OBJECTIVES: The aim of this in vitro study was to compare the demineralization inhibitory effect of gels/solutions used in combination with either standard or highly fluoridated dentifrices on sound dentin as well as on artificial dentin caries-like lesions. METHODS: Bovine dentin specimens (n = 240) with two different surfaces each (sound [ST] and artificial caries lesion [DT]) were prepared and randomly allocated to twelve groups. Weekly interventions during pH-cycling (28 days, 6 × 120 min demineralization/day) were: the application of gels/solutions containing amine fluoride/sodium fluoride (12,500 ppm F [ppm]; pH = 4.4; AmF); NaF (12,500 ppm; pH = 6.6; NaF1); NaF (12,500 ppm; pH = 6.3; NaF2); silver diamine fluoride (14,200 ppm; pH = 8.7; SDF); acidulated phosphate fluoride (12,500 ppm; pH = 3.8; APF), and no intervention (standard control; S). Furthermore, half of the specimens in each group were brushed (10 s; twice per day) with dentifrice slurries containing either 1,450 ppm (e.g., AmF1450) or 5,000 ppm (e.g., AmF5000). Differences in integrated mineral loss (ΔΔZ) and lesion depth (ΔLD) were calculated between values before and after pH-cycling using transversal microradiography. RESULTS: After pH-cycling Ss showed significantly increased ΔZDT and LDDT values, indicating further demineralization. In contrast, except for one, all groups including fluoride gels/solutions showed significantly decreased ΔZDT values. Additional use of most fluoride gels/solutions significantly enhanced mineral gain, mainly in the surface area; however, acidic gels/solutions seemed to have negative effects on lesion depths. SIGNIFICANCE: Under the present pH-cycling conditions the highly fluoridated dentifrice significantly reduced caries progression and additional application of nearly all of the fluoride gels/solutions resulted in remineralization. However, there was no difference in the remineralizing capacity of fluoride gels/solutions when used in combination with either standard or highly fluoridated dentifrices.

The Effect of Solutions Containing Extracts of Vochysia tucanorum Mart., Myrcia bellaCambess., Matricaria chamomilla L. and Malva sylvestris L. on Cariogenic Bacterial Species and Enamel Caries Development.

This study evaluated the effect of experimental solutions containing plant extracts on bacterial species and enamel caries prevention. Microcosm biofilm was produced from human saliva mixed with McBain saliva (0.2% sucrose) on bovine enamel for 5 days (3 days under anaerobiosis and 2 days under aerobiosis) at 37°C. From the 2nd day, the following treatments were applied (1 × 60 s/day): Vochysia tucanorum (10 mg/mL); Myrcia bella (5 mg/mL); Matricaria chamomilla (80 mg/mL); Malva sylvestris, fluoride, and xylitol (Malvatricin Plus®); 0.12% chlorhexidine (CHX, PerioGard®); and PBS (negative control). The medium pH was measured. Quantitative polymerase chain reaction was performed for the detection of Streptococcus mutans and Lactobacillus spp. Enamel demineralization was measured by spectral-domain optical coherence tomography. The data were compared by means of the Kruskal-Wallis/Dunn, two-way ANOVA/Bonferroni, and ANOVA/Tukey tests (p < 0.05). The pH decreased after sucrose exposure; only CHX reestablished pH >5.5 by the last day. CHX also eliminated Lactobacillusspp., but the other treatments did not differ significantly from PBS. Malvatricin Plus® and CHX eliminated S. mutans, but the other treatments did not differ from PBS. Similar results were seen concerning the reduction of lesion depth and reflectivity. The experimental natural-extract solutions were ineffective against cariogenic bacteria and in preventing the development of enamel caries.

Do bleaching gels affect the stability of the masking and caries-arresting effects of caries infiltration-in vitro.

OBJECTIVES: The aim of this study was to evaluate the influence of different bleaching gels on the masking and caries-arresting effects of infiltrated and non-infiltrated stained artificial enamel caries lesions. MATERIALS AND METHODS: Bovine enamel specimens (n = 240) with each two sound areas (SI and SC) and each two lesions (DI and DC) were infiltrated (DI and SI), stained (1:1 red wine-coffee mixture,70 days), and randomly distributed in six groups to be bleached with the following materials: 6%HP (HP-6), 16%CP (CP-16), 35%HP (HP-35), 40%HP (HP-40), and no bleaching (NBl,NBl-NBr). Subsequently, specimens were pH-cycled (28 days, 6 × 60 min demineralization/day) and all groups except NBl-NBr were brushed with toothpaste slurry (1.100 ppm, 2×/day, 10 s). Differences in colorimetric values (ΔL, ΔE) and integrated mineral loss (ΔΔZ) between baseline, infiltration, staining, bleaching, and pH cycling were calculated using photographic and transversal microradiographic images. RESULTS: At baseline, significant visible color differences between DI and SC were observed (ΔEbaseline = 12.2; p < 0.001; ANCOVA). After infiltration, these differences decreased significantly (ΔEinfiltration = 3.8; p < 0.001). Staining decreased and bleaching increased ΔL values significantly (p ≤ 0.001). No significant difference in ΔΔE was observed between before staining and after bleaching (ΔEbleaching = 4.3; p = 0.308) and between the bleaching agents (p = 1.000; ANCOVA). pH-cycling did not affect colorimetric values (ΔEpH-cycling = 4.0; p = 1.000). For DI, no significant change in ΔZ during in vitro period was observed (p ≥ 0.063; paired t test). CONCLUSIONS: Under the conditions chosen, the tested materials could satisfactorily bleach infiltrated and non-infiltrated stained enamel. Furthermore, bleaching did not affect the caries-arresting effect of the infiltration. CLINICAL RELEVANCE: The present study indicates that bleaching is a viable way to satisfactorily recover the appearance of discolored sound enamel and infiltrated lesions.

Analysis of relative bacterial activity and lactate dehydrogenase gene expression of caries-associated bacteria in a site-specific natural biofilm: an ex vivo study.

OBJECTIVES: Detecting bacterial activity is considered a promising approach to monitor shifts from symbiosis to dysbiosis in oral microbiome. The present study aimed at investigating both the relative bacterial activity and the lactate dehydrogenase (ldh) gene expression of caries-associated bacteria in a site-specific natural biofilm. MATERIAL AND METHODS: Sixty subjects (age, mean ± SE: 30.1 ± 1.4) were allocated to two groups: caries-free subjects (CF) or caries-active subjects (CA). CF presented one sound surface (CFS, n = 30). CA presented two donor sites: a cavitated caries lesion (CAC, n = 30) and a sound reference surface (CAS, n = 30). Real-time quantitative PCR (q-PCR) on species or genus level and total bacteria was performed targeting the 16S gene, the 16S rRNA, the ldh gene, and the ldh mRNA (increasing 16S ribosomal RNA copy numbers can function as an indicator of increased energy metabolism). As the 16S rRNA abundance represents the number of ribosomes, while the 16S gene abundance represents the number of genomes, the quotient of the relative abundances functions as a measure for the relative bacterial activity (%). RESULTS: Both lactobacilli and S. mutans showed the highest relative bacterial activity in CAC ((mean ± SE) 218 ± 60% and 61 ± 16%, respectively) and the lowest values for both sound reference surfaces (69 ± 48%; 8 ± 3%). Significant differences were found between CAC and CAS as well as between CAC and CFS for both lactobacilli and S. mutans (p < 0.05). The ldh gene expression of lactobacilli and S. mutans only showed moderate values in CAC (1.90E+03 ± 2.11E+03; 2.08E+04 ± 4.44E+04 transcripts/µl) and CFS (2.04E+03 ± 2.74E+03; 8.16E+03 ± 6.64E+03 transcripts/µl); consequently no significant differences were detected. CONCLUSION AND CLINICAL RELEVANCE: Caries-associated bacteria (lactobacilli and S. mutans) showed the highest relative bacterial activity in plaque of cavitated lesions, the lowest in sound surfaces, allowing the detection of a significant activity shift in health and disease for caries-active patients. However, no significant differences in ldh gene expression could be determined.

Hand-held bioprinting for de novo vascular formation applicable to dental pulp regeneration.

Aim of the study: Deep carious lesions may cause irreversible pulpitis and the current endodontic treatment typically removes the whole dental pulp tissue, which finally reduces lifespan of the teeth. Nowadays, the most frequent treatment is based on removing the infected tissue and filling the root canal with inert synthetic materials. Tissue engineering approaches are important alternatives to the current treatment, because they can potentially maintain the biological function of the tooth instead of sacrificing it.Materials and Methods: In this study, we propose a tissue engineering approach based on a hand-held in situ bioprinting strategy. Our approach enabled bioprinting of cell-loaded collagen-based bioinks with suitable rheological, structural and biological properties, which allowed for vasculogenesis in the root canal.Results: The rheological properties of the bioprintable bioink were measured by oscillatory amplitude sweep testing and were corroborated by macroscopic evaluation after in vitro culture, in which printed bioinks maintained their original form without contraction. Moreover, we showed evidence for successful vasculogenesis in bioprintable bioinks with comparable quality and quantity to control fibrin and collagen non-bioprintable hydrogels.Conclusions: We conclude that hand-held bioprinting holds potential for in situ treatment of dental diseases with successful evidence for vascular tube formation, as an asset for maintenance of the biological function of the tooth.

Effects of Dentifrices Differing in Fluoride Content on Remineralization Characteristics of Dentin in vitro.

OBJECTIVES: The aim of this study was to compare the caries preventive effect of highly fluoridated dentifrices and gels on sound dentin as well as on artificial dentin caries-like lesions. METHODS: Bovine dentin specimens (n = 240), with 2 different surfaces each (1 sound surface [sound treatment (ST)] and one caries lesion [demineralized treatment (DT)]), were prepared and randomly allocated to one highly (6 × 120 min demineralization/day [H]) and one lowly cariogenic (6 × 60 min demineralization/day [L]) pH-cycling model. Treatments during pH-cycling (28 days) were: brushing 2×/day with: 0 ppm F [H0/L0], 1,450 ppm F [H1,450/L1,450], 2,800 ppm F [H2,800/L2,800], 5,000 ppm F [H5,000/L5,000], 5,000 ppm F plus TCP [H5,000+TCP/L5,000+TCP], and 12,500 ppm F [H12,500/L12,500] containing dentifrices/gels. Dentifrice/gel slurries were prepared with deionized water (1:2 wt/wt). Differences in integrated mineral loss (∆∆Z) and ∆ lesion depth were calculated between values before and after pH-cycling using transversal microradiography. RESULTS: The correlation between ΔΔZDT and F- was strong for the highly (rH = 0.691; p < 0.001) and moderate (rL = 0.500; p < 0.001) for the lowly cariogenic model, indicating a fluoride dose-response for both. Significant differences for ΔΔZDT and ΔΔZST could be found between H0, H1,450, H5,000, and H12,500 as well as L0, L5,000, and L125,000 (p ≤ 0.046; analysis of covariance [ANCOVA]). Except for 0 ppm F-, no significant difference in ΔΔZST and ΔΔZDT could be found between the highly and lowly cariogenic model (p ≥ 0.056; ANCOVA). CONCLUSION: For both pH-cycling conditions a dose-response for fluoride could be revealed. For elderly people with exposed root surfaces, the use of gels containing 12,500 ppm F instead of regularly (1,450 ppm F) or highly (5,000 ppm F) fluoridated dentifrices should be further investigated, as it offered higher caries-preventive effects in vitro.

Correlation between relative bacterial activity and lactate dehydrogenase gene expression of co-cultures in vitro.

OBJECTIVES: The present study aims at correlating the relative bacterial activity with the H+ concentration and the ldh expression of caries-associated bacteria in co-cultures. MATERIALS AND METHODS: Well plates were prepared with BHI medium and cultures of Lactobacillus paracasei and Fusobacterium nucleatum. Bacterial growth at 37 °C was measured using a microplate-photometer before and after adding sucrose to the samples. Samples of co-cultures (n = 12) and single-species cultures (n = 3) were taken and pH was assessed. Real-time quantitative PCRs were applied targeting the 16S-gene, the 16S-rRNA, the ldh-gene, and the ldh-mRNA. RESULTS: For L. paracasei with sucrose, an increase in relative bacterial activity (62.8% ± 23.5% [mean, SE]) was observed, while F. nucleatum showed a clear decrease in relative bacterial activity (- 35.0% ± 9.6%). Simultaneously, the H+ concentration increased (1.15E-05 mol*l-1 ± 4.61E-07 mol*l-1). Consequently, a significant positive correlation was found between L. paracasei's relative bacterial activity and H+ concentration (Spearman rank correlation, r = 0.638; p = 0.002), while F. nucleatum exhibited a negative correlation (r = - 0.741; p ≤ 0.001). Furthermore L. paracasei with sucrose showed a moderate, but significant positive correlation between relative bacterial activity and ldh-expression (r = 0.307; p = 0.024). CONCLUSIONS AND CLINICAL RELEVANCE: The relative bacterial activity after sucrose pulse showed a significant correlation not only to the acid production (H+ concentration) but also to ldh expression of L. paracasei. However, further research is required to confirm these findings in a mature biofilm in vivo.

Randomized in situ study on the efficacy of CO2 laser irradiation in increasing enamel erosion resistance.

OBJECTIVES: The aim of this double-blind, randomized in situ study was to evaluate the erosion-preventive effect of a specific set of CO2 laser parameters, associated or not with fluoride. METHODS: Two hundred forty bovine enamel blocks were prepared for individual palatal appliances (n = 6 samples/appliance). The study had four phases of 5 days each, with ten volunteers and the following treatments: CO2 laser irradiation (L), fluoride treatment (F), combined fluoride and laser treatment (FL), and no treatment, control (C). Laser irradiation was performed at 0.3 J/cm2 (5 µs/226 Hz/10.6 µm) and the fluoride gel contained AmF/NaF (12'500 ppm F-/pH = 4.8-6). For erosive demineralization, the appliances were immersed extra-orally in citric acid (0.05 M/20 min/pH = 2.3) twice daily. Analysis of enamel surface loss was done using a 3D-laser profilometer on 3 days. Additionally, fluoride uptake was quantified and scanning electron microscopies were done. Data were analyzed with repeated measures ANOVA and post hoc pairwise comparisons (α = 0.05). RESULTS: At all analyzing days, both laser groups caused the lowest means of enamel loss, which were also statistically significant lower than C (p < 0.05). At day 5, FL means ± SD (33.6 ± 12.6 µm) were even significantly lower than all other groups (C 67.8 ± 15.4 µm; F 57.5 ± 20.3 µm; L 46.8 ± 14.5 µm). Significantly increased enamel fluoride uptake was observed for both fluoride-containing groups (p < 0.05) at day 1. CONCLUSION: Compared to the control, the CO2 laser irradiation with a specific set of laser parameters (0.3 J/cm2/5 µs/226 Hz) either alone or in combination with a fluoride gel (AmF/NaF) could significantly decrease enamel erosive loss up to 5 days in situ. CLINICAL RELEVANCE: Combined CO2 laser-fluoride treatment has a significant anti-erosive effect.

Caries-Preventive Effect of NaF, NaF plus TCP, NaF plus CPP-ACP, and SDF Varnishes on Sound Dentin and Artificial Dentin Caries in vitro.

The aim of this study was to compare the caries-preventive effect of different fluoride varnishes on sound dentin as well as on artificial dentin caries-like lesions. Bovine dentin specimens (n = 220) with one sound surface (ST) and one artificial caries lesion (DT) were prepared and randomly allocated to 11 groups. The interventions before pH cycling were as follows: application of a varnish containing NaF (22,600 ppm F-; Duraphat [NaF0/NaF1]), NaF plus tricalcium phosphate (22,600 ppm F-; Clinpro White Varnish Mint [TCP0/TCP1]), NaF plus casein phosphopeptide-stabilized amorphous calcium phosphate complexes (CPP-ACP; 22,600 ppm F-; MI Varnish [CPP0/CPP1]), or silver diamine fluoride (SDF; 35,400 ppm F-; Cariestop 30% [SDF0/SDF1]) and no intervention (NNB/N0/N1). During pH cycling (14 days, 6 × 120 min demineralization/day) half of the specimens in each group were brushed (10 s; 2 times/day) with either fluoride-free ("0"; e.g., TCP0) or 1,100 ppm F- ("1"; e.g., TCP1) dentifrice slurry. In another subgroup, the specimens were pH cycled but not brushed (NNB). Differences in integrated mineral loss (ΔΔZ), lesion depth (ΔLD), and colorimetric values (ΔΔE) were calculated between the values after initial demineralization and those after pH cycling, using transversal microradiography and photographic images. After pH cycling, no discoloration could be observed. Furthermore, NNB, N0, and N1 showed significantly increased ΔZDT/LDDT and ΔZST/LDST values, indicating further demineralization. In contrast, CPP0, CPP1, SDF0, and SDF1 showed significantly decreased ΔZDT/LDDT values, indicating remineralization (p ≤ 0.004; paired t test). CPP0, CPP1, SDF0, and SDF1 showed significantly higher changes in ΔΔZDT/ΔLDDT and ΔΔZST/ΔLDST than NNB, N0, and N1 (p < 0.001; Bonferroni post hoc test). In conclusion, under the conditions chosen, all fluoride varnishes prevented further demineralization. However, only NaF plus CPP-ACP and SDF could remineralize artificial dentin caries-like lesions under net-demineralizing conditions, thereby indicating that NaF plus CPP-ACP and SDF may be helpful to high-caries-risk patients.

Effects of Self-Assembling Peptide P11-4, Fluorides, and Caries Infiltration on Artificial Enamel Caries Lesions in vitro.

The application of a self-assembling peptide on noncavitated caries lesions is supposed to be a feasible approach to facilitate remineralization and mask their unfavorable appearance. However, demineralizing conditions are common in the oral environment, so the aim of this pH-cycling study was to compare recommended and novel treatment methods regarding their ability to hamper demineralization and as a consequence mask artificial enamel caries lesions. Artificial caries lesions were prepared in bovine enamel and randomly allocated to 11 groups (n = 22). Treatments before pH-cycling were as follows: the application of a self-assembling peptide (Curodont™ Repair [C]), a low-viscosity resin (Icon® [I]), 2 fluoride solutions (10,000 ppm F-: Elmex fluid [E] and 43,350 ppm F-: Tiefenfluorid® [T]), and no intervention (N). During pH-cycling (28 days, 6 × 60 min demineralization/day) half of the specimens in each group were brushed (10 s; 2 ×/day) with either fluoride-free (named e.g., C0) or NaF (1,100 ppm F-; e.g., C1) dentifrice slurry. In another subgroup specimens were pH-cycled but not brushed (NNB). Differences in integrated mineral loss (ΔΔZ), lesion depth (ΔLD), and colorimetric values (ΔΔE) were calculated between values after pre-demineralization, surface treatment, and pH-cycling. Specimens of C0, C1, NNB, N0, N1, T0, and E0 showed significantly increased ΔZ and LD values after pH-cycling (p ≤ 0.003; paired t test). C0, C1, NNB, and N0 showed significantly higher changes in ΔΔZ than E1, I0, I1, and T1 (p < 0.001; ANOVA). Significantly reduced colorimetric values could only be observed for I1, I0, E1, and E0 after treatment and after pH-cycling (p ≤ 0.027; paired t test). In conclusion, under the conditions chosen only the application of a low-viscosity resin could mask caries lesions significantly, whereas self-assembling peptides could neither inhibit lesion progression nor mask the lesions considerably.

Impact of combined CO2 laser irradiation and fluoride on enamel and dentin biofilm-induced mineral loss.

OBJECTIVES: The caries-protective effects of CO2 laser irradiation on dental enamel have been demonstrated using chemical demineralization models. We compared the effect of CO2 laser irradiation, sodium fluoride, or both on biofilm-induced mineral loss (∆Z) and Streptococcus mutans adhesion to enamel and dentin in vitro. MATERIALS AND METHODS: Ground, polished bovine enamel, and dentin samples were allocated to four groups (n = 12/group): no treatment (C); single 22,600-ppm fluoride (F) varnish (5 % NaF) application; single CO2 laser treatment (L) with short pulses (5 µs/λ = 10.6 µm); and laser and subsequent fluoride treatment (LF). Samples were sterilized and submitted to an automated mono-species S. mutans biofilm model. Brain heart infusion plus 5 % sucrose medium was provided eight times daily, followed by rinses with artificial saliva. After 10 days, bacterial numbers in biofilms were enumerated as colony-forming units/ml (CFU/ml) (n = 7/group). ∆Z was assessed using transversal microradiography (n = 12/group). Univariate ANOVA with post hoc Tukey honestly-significant-difference test was used for statistical analysis. RESULTS: Bacterial numbers were significantly higher on dentin than enamel (p < 0.01/ANOVA). On dentin, LF yielded significantly lower CFUs than other groups (p = 0.03/Tukey), while no differences between groups were found for enamel. The lowest ∆Z in enamel was observed for L (mean/SD 2036/1353 vol%×µm), which was not only significantly lower than C (9642/2452 vol%×µm) and F (7713/1489 vol%×µm) (p < 0.05) but also not significantly different from LF (3135/2628 vol%×µm) (p > 0.05). In dentin, only LF (163/227) significantly reduced ∆Z (p < 0.05). CONCLUSION/CLINICAL RELEVANCE: CO2 laser irradiation did not increase adhesion of S. mutans in vitro. Laser treatment alone protected enamel against biofilm-induced demineralization, while a combined laser-fluoride application was required to protect dentin.

Re- and Demineralization Characteristics of Enamel Depending on Baseline Mineral Loss and Lesion Depth in situ.

OBJECTIVES: The aim of this double-blinded, randomized, cross-over in situ study was to evaluate the re- and demineralization characteristics of sound enamel as well as lowly and highly demineralized caries-like enamel lesions after the application of different fluoride compounds. METHODS: In each of three experimental legs of 4 weeks, 21 participants wore intraoral mandibular appliances containing 4 bovine enamel specimens (2 lowly and 2 highly demineralized). Each specimen included one sound enamel and either one lowly demineralized (7 days, pH 4.95) or one highly demineralized (21 days, pH 4.95) lesion, and was positioned 1 mm below the acrylic under a plastic mesh. The three randomly allocated treatments (application only) included the following dentifrices: (1) 1,100 ppm F as NaF, (2) 1,100 ppm F as SnF2 and (3) 0 ppm F (fluoride-free) as negative control. Differences in integrated mineral loss (x0394;x0394;Z) and lesion depth (x0394;LD) were calculated between values before and after the in situ period using transversal microradiography. RESULTS: Of the 21 participants, 6 did not complete the study and 2 were excluded due to protocol violation. Irrespectively of the treatment, higher baseline mineral loss and lesion depth led to a less pronounced change in mineral loss and lesion depth. Except for x0394;x0394;Z of the dentifrice with 0 ppm F, sound surfaces showed significantly higher x0394;x0394;Z and x0394;LD values compared with lowly and highly demineralized lesions (p < 0.05, t test). CONCLUSION: Re- and demineralization characteristics of enamel depended directly on baseline mineral loss and lesion depth. Treatment groups should therefore be well balanced with respect to baseline mineral loss and lesion depth.

Analysis of Bacterial Activity in Sound and Cariogenic Biofilm: A Pilot in vivo Study.

Dental caries is a multifactorial disease with many associated microbial taxa, but only a few are notably contributing to acidogenicity. The ribosome number and the corresponding 16S ribosomal RNA (rRNA) concentration are considered a molecular indicator for general metabolic activity of bacteria, as they are elevated with increased anabolic and catabolic activities. We hypothesize that the activity of aciduric/acidogenic bacterial taxa, reflected by a rise in ribosomal counts, could resolve differences between plaque biofilm from sound surfaces and caries lesions. The included subjects were allocated to two groups: caries-free (CF) or caries-active (CA). CF subjects presented one donor site, namely one sound surface (CFS, n = 10), whereas CA subjects presented two donor sites: a cavitated lesion with an ICDAS score of 5-6 (CAC, n = 13), and a sound reference surface (CAS, n = 13). Four aciduric/acidogenic bacterial taxa (Streptococcus mutans, lactobacilli, Bifidobacterium dentium, and Scardovia wiggsiae) and one asaccharolytic taxon (fusobacteria) as a contrast were selected. 16S rRNA and 16S rRNA genes were quantified by quantitative PCR. Based on these parameters, bacterial and ribosomal counts, as well as relative activities were calculated as the quotient of relative ribosomal abundance and relative genome abundance. Caries-associated bacteria showed the highest relative activity in caries lesions (e.g. lactobacilli CAC: 177.5 ± 46.0%) and lower activities on sound surfaces (e.g. lactobacilli CAS: 96.3 ± 31.5%), whereas asaccharolytic fusobacteria were most active on sound surfaces and less active in caries lesions (CFS: 275.7 ± 171.1%; CAS: 205.8 ± 114.3%; CAC: 51.1 ± 19.0%). Thus, the present study suggests different activity patterns for biofilms from CF and CA individuals.