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Item Age effect on presence, susceptibility and treatment of erosive tooth wear(2018) Algarni, Amnah Abdullah; Hara, Anderson T.; Lippert, Frank; Platt, Jeffrey A.; González-Cabezas, Carlos; Ungar, PeterErosive tooth wear (ETW) is a growing dental condition often associated with aging. This in-vitro project comprised three studies aiming to investigate the impact of tooth age on ETW susceptibility and prevention. In the first study, un-identified extracted premolars were collected and had their ages estimated using validated dental forensic methods. The premolars were examined to investigate the relationship between age and presence and severity of ETW, as well as other main dental-hard tissues conditions. ETW, dental caries, fluorosis, extrinsic staining and tooth color were evaluated using established clinical indices. In the second study, the tooth age impact on ETW susceptibility and response to preventive treatments (Sn+F, NaF, and de-ionized water control) were evaluated using representative samples from the initial study. Enamel and dentin specimens were prepared and subjected to daily erosion-treatmentremineralization cycling procedure. Surface loss (SL) was determined during and after the cycling, by optical profilometry. Similar protocol was adopted in the third study with the addition of toothbrushing abrasion to the model, in order to explore the interplay between age and toothpaste abrasivity on erosion-abrasion development. SL was measured during and after the erosion-toothbrushing-remineralization cycling. The relationships between age and the investigated variables were assessed using linear regression models. In conclusion: 1. The presence and severity of ETW, dental caries, and extrinsic staining increased with age, while of enamel fluorosis decreased. Tooth also showed to be darker with age. 2. Susceptibility of enamel and dentin to demineralization increased with age. Sn+F showed the highest anti-erosive efficacy, and was not affected by age. NaF showed lower efficacy on dentin, which increased with age. 3. Enamel and dentin SL increased with toothpaste abrasivity level. Dentin SL also increased with age. Age effect on enamel SL was observed only with low abrasive toothpaste. Age-related changes on enamel and dentin affected ETW development.Item Artificial Caries Lesion Characteristics after Secondary Demineralization with Theobromine-Containing Protocol(MDPI, 2021-01-08) Nassar, Hani M.; Lippert, Frank; Cariology, Operative Dentistry and Dental Public Health, School of DentistryDeveloping artificial caries lesions with varying characteristics is needed to adequately study caries process in vitro. The objective of this study was to investigate artificial caries lesion characteristics after secondary demineralization protocol containing theobromine and fluoride. Sixty bovine enamel slabs (4 × 3 mm) were demineralized using a Carbopol-containing protocol for 6 days. A baseline area (2 × 3 mm) was protected with acid-resistant nail varnish, after which specimens were exposed for 24 h to a secondary demineralization protocol containing acetic acid plus one of four fluoride/theobromine combinations (n = 15): theobromine (50 or 200 ppm) and fluoride (0 or 1 ppm). Specimens were sectioned and analyzed using transverse microradiography for changes in mineral content, lesion depth, and surface layer mineralization. Data was analyzed using paired t-test and analysis of variance followed by Bonferroni test at 0.05 significance level. After secondary demineralization, fluoride-containing groups had significantly deeper lesions (p = 0.002 and 0.014) compared to the group with 0 ppm fluoride and 50 ppm theobromine. Mineral content and lesion depth were significantly different compared to baseline for all groups. Theobromine did not show an added effect on mineral uptake. Theobromine-containing groups exhibited particularly deep lesions with a more uniform mineral profile in the presence of fluoride.Item Detection of artificial enamel caries-like lesions with a blue hydroxyapatite-binding porosity probe(Elsevier, 2023-08) Lippert, Frank; Eder, Jennifer S.; Eckert, George J.; Mangum, Jonathan; Hegarty, Kerry; Cariology, Operative Dentistry and Dental Public Health, School of DentistryObjectives This in vitro study investigated the ability of a blue protein-based hydroxyapatite porosity probe to selectively detect artificial enamel caries-like lesions of varying severities. Methods Artificial caries-like lesions were formed in enamel specimens using a hydroxyethylcellulose-containing lactic acid gel for 4/12/24/72 or 168 h. One untreated group was used as a control. The probe was applied for 2 min and unbound probe rinsed off with deionized water. Surface color changes were determined spectrophotometrically (L*a*b* color space) and with digital photography. Lesions were characterized using quantitative light-induced fluorescence (QLF), Vickers surface microhardness, and transverse microradiography (TMR). Data were analyzed using one-way ANOVA. Results Digital photography did not reveal any discoloration in unaffected enamel. However, all lesions stained blue with color intensity positively correlated with demineralization times. The color data reflected similar trends: lesions became significantly darker (L* decreased) and bluer (b* decreased), while overall color differences (ΔE) increased significantly after probe application (4-h lesion, mean±standard deviation: ΔL*=-2.6 ± 4.1/Δb*=0.1 ± 0.8/ΔE=5.5 ± 1.3 vs. 168-h lesion: ΔL*=-17.3 ± 1.1/Δb*=-6.0 ± 0.6/ΔE=18.7 ± 1.1). TMR analysis revealed distinct differences in integrated mineral loss (ΔZ) and lesion depth (L) between demineralization times (4-h lesion: ΔZ=391±190 vol%min × µm/L = 18.1 ± 10.9 µm vs. 168-h lesion: ΔZ=3606±499 vol%min × µm/L = 111.9 ± 13.9 µm). QLF and microhardness were also able to differentiate between demineralization times. L and ΔZ strongly correlated (Pearson correlation coefficient [r]) with Δb* (L vs. Δb*: r=-0.90/ΔZ vs. Δb*: r=-0.90), ΔE (r = 0.85/r = 0.81), and ΔL* (r=-0.79/r=-0.73). Conclusion Considering the limitations of this study, the blue protein-based hydroxyapatite-binding porosity probe appears to be sufficiently sensitive to distinguish between unaffected enamel and artificial caries-like lesions. Clinical significance Early detection of enamel caries lesions remains one of the most critical aspects in the diagnosis and management of dental caries. This study highlighted the potential of a novel porosity probe in detecting artificial caries-like demineralization by objective means.Item Effect of Bottled Water and Fluoride Toothpaste Usage on Caries Lesion Remineralization(2023) Qaw, Masoumah Samir; Lippert, Frank; Al Dehailan, Laila; Hara, Anderson T; Cook, N. BlaineBackground: The importance of fluoride in the prevention of dental caries has been well documented in the literature, as it inhibits demineralization of the tooth structure and enhances remineralization. One of the major public health policies to prevent caries is to provide the population with an adequate amount of fluoride through community water fluoridation. Nowadays many people drink bottled water instead of tap water due to its easy access, convenience, and low cost. Besides fluoride, other minerals present in tap and bottled water, such as calcium and magnesium, are also important in decreasing dental caries prevalence. However, our knowledge of the role of bottled water in caries prevention and especially when combined with fluoride toothpaste usage is still poor. Objectives: The aim of this in-vitro study was to evaluate the effects of some bottled waters on fluoride toothpaste efficacy in enhancing caries lesion remineralization. Methodology: Early caries lesions were created in bovine enamel specimens and stratified into treatment groups based on Vickers surface microhardness (VHN). The present study followed a two (fluoride and fluoride-free toothpaste) by five (four bottled waters and tap water) factorial design. The treatment groups were bottled water with the following attributes: a) 309.9 ppm Ca/1.20 ppm F; b) 118.4 ppm Ca/0.16 ppm F; c) 1.00 ppm Ca/1.01 ppm F; d) 0.1 ppm Ca/0.04 ppm F, and tap water (48.7 ppm Ca/0.7 ppm F). The five water groups were paired either with 1100 ppm fluoride or fluoride-free toothpaste, yielding 10 groups. Specimens were pH-cycled for 10 days with the daily regimen comprised of twice daily toothpaste slurry, with four exposures to water in between. VHN was measured, again, and the difference calculated (ΔVHN). Data were analyzed using two-way ANOVA at a 5-percent significance level. Results: The two-way interaction between water and toothpaste was significant (p < 0.001). All groups except fluoride-free toothpaste/bottled water with 0.1 ppm Ca/0.04 ppm F (p = 0.411) had significant increases in VHN after pH cycling (p ≤ 0.023). Fluoridated toothpaste resulted in a higher rate of remineralization compared to fluoridefree toothpaste (all p < 0.001). Bottled water with 1.20 ppm F/309.9 ppm Ca exhibited the greatest extent of remineralization within fluoride toothpaste groups (p < 0.001) and higher remineralization than lower fluoride water in fluoride-free toothpaste groups (p ≤ 0.006). Within the fluoridated toothpaste group, tap water exhibited significantly less remineralization compared to all bottled waters (all p < 0.001). Conclusion: Within the limitation of this study, bottled water with higher fluoride and calcium concentrations might improve fluoridated toothpaste efficacy by enhancing remineralization of early enamel caries-like lesions.Item Farnesol Delivery via Polymeric Nanoparticle Carriers Inhibits Cariogenic Cross-Kingdom Biofilms and Prevents Enamel Demineralization(Wiley, 2022) Ito, Tatsuro; Sims, Kenneth R., Jr.; Liu, Yuan; Xiang, Zhenting; Arthur, Rodrigo A.; Hara, Anderson T.; Koo, Hyun; Benoit, Danielle S. W.; Klein, Marlise I.; Biomedical and Applied Sciences, School of DentistryStreptococcus mutans and Candida albicans are frequently detected together in the plaque from patients with early childhood caries (ECC) and synergistically interact to form a cariogenic cross-kingdom biofilm. However, this biofilm is difficult to control. Thus, to achieve maximal efficacy within the complex biofilm microenvironment, nanoparticle carriers have shown increased interest in treating oral biofilms in recent years. Here, we assessed the anti-biofilm efficacy of farnesol (Far), a hydrophobic antibacterial drug and repressor of Candida filamentous forms, against cross-kingdom biofilms employing drug delivery via polymeric nanoparticle carriers (NPCs). We also evaluated the effect of the strategy on teeth enamel demineralization. The farnesol-loaded NPCs (NPC+Far) resulted in a 2-log CFU/mL reduction of S. mutans and C. albicans (hydroxyapatite disc biofilm model). High-resolution confocal images further confirmed a significant reduction in exopolysaccharides, smaller microcolonies of S. mutans, and no hyphal form of C. albicans after treatment with NPC+Far on human tooth enamel (HT) slabs, altering the biofilm 3D structure. Furthermore, NPC+Far treatment was highly effective in preventing enamel demineralization on HT, reducing lesion depth (79% reduction) and mineral loss (85% reduction) versus vehicle PBS-treated HT, while NPC or Far alone had no differences with the PBS. The drug delivery via polymeric NPCs has the potential for targeting bacterial-fungal biofilms associated with a prevalent and costly pediatric oral disease, such as ECC.Item Improving Enamel Acid Resistance by an Intraoral Fluoride-Release Device Incorporating Cationic Hydroxy Cellulose Gel Using 3D Printer Molding(MDPI, 2024-11-23) Satou, Ryouichi; Odaka, Kento; Sako, Ryo; Ando, Masatoshi; Sugihara, Naoki; Biomedical and Applied Sciences, School of DentistryAn intraoral fluoride-releasing device (IFRD) is a cost-effective tool for introducing fluoride into the oral cavity. It allows prolonged uptake of low concentrations of fluoride into teeth. We developed a new IFRD using 3D additive manufacturing and a new low-release fluoride gel. Gels for IFRDs were synthesized from hydroxyethyl cellulose (SE600) and cationic hydroxyethyl cellulose (L200). We compared the effects of the new cationic fluoride slow-release gel and non-cationic gel on enamel acid resistance in vitro. Cationization significantly increased fluoride ion concentration, as evident from its concentrations of 0.68 ± 0.08 ppm for SE600 and 4.24 ± 0.83 ppm for L200 after 60 min of immersion in distilled water. In addition, the acid resistance of bovine tooth enamel post-application was analyzed by measuring tooth loss, mineral loss (ΔZ), and lesion depth (Ld) using polarized light microscopy, electron microscopy, and micro-radiography. Compared to the SE600 group, the cationic L200 group had significantly reduced ΔZ and Ld, enhancing enamel acid resistance. This device could be implemented in areas where adequate oral care is challenging, including preventive dentistry, ward management, nursing homes, and dental clinic visits.Item The influence of baseline hardness and chemical composition on enamel demineralization and subsequent remineralization(2017-05-09) Alkattan, Rana; Ando, Masatoshi; Lippert, Frank; Cook, Norman BlaineBACKGROUND Several studies have reported that harder enamel with higher contents of calcium (Ca), phosphorus (P) and fluorine (F) coupled with lower contents of carbonate (C), magnesium (Mg) and nitrate (N) was found to be more resistant to demineralization. Additionally, the hardness of dental enamel was found to have a strong correlation with its chemical content. However, yet to be established is the relation between the physical and chemical structure of enamel and its response to de- and remineralizing conditions. OBJECTIVES The aims of this laboratory study were: 1) To investigate the hardness and chemical content of sound enamel and their influence on demineralization; 2) To investigate these properties in demineralized enamel and their influence on remineralization; and 3) To investigate these properties in sound enamel and their influence on remineralization. MATERIALS AND METHODS Incipient subsurface caries lesions were created in 94 bovine enamel specimens using Carbopol C907 using three demineralization times. The specimens were then pH-cycled and treated using either 367 ppm F sodium fluoride or a placebo. Knoop surface microhardness (SMH), Energy dispersive X-ray spectroscopy (EDS) and Transverse microradiography (TMR) were performed on the specimens at all stages and compared between them. TMR variables included integrated mineral loss (ΔZ), Lesion depth (L) and maximum mineral density of the surface zone (SZmax). Data were analyzed using three- and four-way ANOVA and Pearson correlation coefficients were calculated. RESULTS SMH, ΔZ, L and SZmax were significantly different among stages, demineralization times and treatment. The weight% of F at the surface was significantly affected by treatment, irrespective of demineralization time. A statistically significant moderate correlation was found between SZmax and ΔZ and SZmax and L after pH cycling. SMH also correlated weakly to moderately with TMR data. CONCLUSIONS SMH and SZmax decreased while ΔZ and L increased with increased demineralization time. Both fluoride and non-fluoride specimens were able to remineralize, which emphasizes the role of saliva in mineralization. The Ca:P ratio remained stable at various stages, indicating the stoichiometric dissolution and redeposition of minerals. The greatest deposition of F was at the surface and its increase led to an increase in SMH and SZmax. SMH values showed that harder specimens at baseline and after demineralization remained hard after demineralization and pH-cycling, respectively, although this correlation was weak. Additionally, harder lesions showed less L and ΔZ and greater SZmax. RELEVANCE This in-vitro study will help better understand the caries process and the impact of physical and chemical characteristics of enamel on de- and remineralization challenges.Item Monitoring of Simulated Erosive Tooth Wear by Cross-Polarization Optical Coherence Tomography(2019-06) Alghilan, Maryam Abdulkareem; Hara, Anderson T.; Lippert, Frank; Platt, Jeffrey A.; González-Cabezas, Carlos; Fried, DanielErosive tooth wear (ETW) is an emerging dental condition manifested clinically as tooth surface loss, eventually impairing the teeth’s structural integrity, function, and esthetics. Both research and practice are in need of a quantitative, non-destructive method to monitor ETW. Cross-polarization optical coherence tomography (CP-OCT), an advanced imaging tool, shows great potential to fulfill this need, but its feasibility and shortcomings remain unclear. In this dissertation, I explored the capability of CP-OCT to monitor ETW in three in vitro studies, one per chapter. Chapter 2 investigated the effects of enamel surface roughness and dental erosion severity on CP-OCT dental surface loss measurements. Chapter 3 tested the effects of enamel surface roughness and dental erosion on CP-OCT enamel thickness measurements at different simulated wear levels. Chapter 4 explored the ability of CP-OCT to quantify the thickness of natural and wornout enamel surfaces and to estimate longitudinally the wear depths resulting from simulated wear. I concluded: (1) enamel surface roughness did not affect CP-OCT measurements of enamel surface loss, however, the estimated error limited the appropriate assessment of the initial stages of dental erosion surface loss using CP-OCT; (2) enamel surface roughness and dental erosion did not affect CP-OCT enamel thickness measurements, and the CP-OCT differentiated the simulated enamel wear levels; and (3) CP-OCT quantified thickness of natural enamel before, during, and after the tooth wear simulation and allowed wear depth estimation following the simulated wear.Item Relative fluoride response of caries lesions created in fluorotic and sound teeth studied underremineralizing conditions(2013) Alhawij, Hala M.; Martilnez-Mier, Esperanza de los A. (Esperanza de los Angeles), 1967-; Lippert, Frank; Cochran, Michael A. (Michael Alan), 1944-; Soto-Rojas, Armando E.; Zandoná, Andréa G. Ferreira (Andréa Gonçalves Ferreira),1969-Fluoride prevents caries by decreasing demineralization and promoting remineralization of enamel crystals during lesion formation and progression. The chemical, morphological, and histologic characteristics of teeth with fluorosis may alter the response of enamel to fluoride during the caries process. However, the effectiveness of fluoride in fluorosed teeth has not been thoroughly studied. Therefore, the current study utilized an in-vitro net remineralization model to study differences between fluorosed and sound enamel with regards to caries lesion remineralization and progression. Hypotheses: There is no significant difference in the effectiveness of fluoride to enhance caries lesion remineralization between fluorosed and non-fluorosed (sound) teeth after changes in enamel fluorescence relative to the lesion baseline. Objectives: The primary objective is to investigate the relative fluoride response of caries lesions created in sound and fluorosed teeth of varying severities under remineralizing conditions in vitro. Secondary objectives: a) To investigate the impact of the presence and severity of enamel fluorosis on caries lesion formation, and b) To investigate the acquired acid resistance of pH-cycled lesions relative to their fluorosis severity. Materials and Methods: Three hundred sixty (360) extracted human molars (sound and fluorosed) were collected and divided into four main groups based on their TF score (0; 1; 2; 3), with sample size of n = 45 per group. Each group was divided into two subgroups: no fluoride/placebo and fluoride/treatment. Artificial caries lesions were created on the teeth and pH-cycled for 20 days using a net remineralization model. The lesions were assessed to determine differences in acquired acid resistance. Quantitative light-induced fluorescence (QLF) was used throughout the study to determine changes in enamel fluorescence relative to a sound enamel window, thereby providing information about enamel de- or remineralization. The results of this study demonstrated that enamel fluorosis levels had a significant effect on lesion remineralization after pH cycling (p > 0.05). The teeth with moderate dental fluorosis had a significantly better response to remineralization and more resistance to a subsequent acid challenge than sound teeth in the presence of fluoride.