Amelogenesis: Transformation of a protein-mineral matrix into tooth enamel.

During enamel formation, the organic enamel protein matrix interacts with calcium phosphate minerals to form elongated, parallel, and bundled enamel apatite crystals of extraordinary hardness and biomechanical resilience. The enamel protein matrix consists of unique enamel proteins such as amelogenin, ameloblastin, and enamelin, which are secreted by highly specialized cells called ameloblasts. The ameloblasts also facilitate calcium and phosphate ion transport toward the enamel layer. Within ameloblasts, enamel proteins are transported as a polygonal matrix with 5 nm subunits in secretory vesicles. Upon expulsion from the ameloblasts, the enamel protein matrix is re-organized into 20 nm subunit compartments. Enamel matrix subunit compartment assembly and expansion coincide with C-terminal cleavage by the MMP20 enamel protease and N-terminal amelogenin self-assembly. Upon enamel crystal precipitation, the enamel protein phase is reconfigured to surround the elongating enamel crystals and facilitate their elongation in C-axis direction. At this stage of development, and upon further amelogenin cleavage, central and polyproline-rich fragments of the amelogenin molecule associate with the growing mineral crystals through a process termed "shedding", while hexagonal apatite crystals fuse in longitudinal direction. Enamel protein sheath-coated enamel "dahlite" crystals continue to elongate until a dense bundle of parallel apatite crystals is formed, while the enamel matrix is continuously degraded by proteolytic enzymes. Together, these insights portrait enamel mineral nucleation and growth as a complex and dynamic set of interactions between enamel proteins and mineral ions that facilitate regularly seeded apatite growth and parallel enamel crystal elongation.

El esmalte dental bovino como modelo experimental para la investigación en odontología: una revisión de la literatura / Bovine dental enamel as experimental model for research in dentistry. A literature review

El propósito de esta revisión bibliográfica es aportar información actualizada acerca de las características de los dientes bovinos en relación con su uso como sustitutos de dientes humanos en trabajos de investigación. De acuerdo con la información registrada, los dientes bovinos serían excelentes sustitutos de la dentición humana para la realización de ensayos de laboratorio con el esmalte dental como modelo experimental (AU)

The purpose of this bibliographic review is to provide updated information about the characteristics of bovine teeth to be used as substitutes for human teeth in dental research. According to the information recorded, bovine teeth appear to be excellent substitutes for human dentition for conducting laboratory tests, using dental enamel as an experimental model (AU)

Enamel-like tissue regeneration by using biomimetic enamel matrix proteins.

Enamel regeneration currently -is limited by our inability to duplicate artificially its complicated and well-aligned hydroxyapatite structure. The initial formation of enamel occurs in enamel organs where the ameloblasts secret enamel extracellular matrix formed a unique gel-like microenvironment. The enamel extracellular matrix is mainly composed by amelogenin and non-amelogenin. In this study, an innovative strategy was proposed to regenerate enamel-like tissue by constructing a microenvironment using biomimetic enamel matrix proteins (biomimetic EMPs) composed of modified leucine-rich amelogenin peptide (mLRAP) and non-amelogenin analog (NAA). Impressively, the regenerated enamel in this biomimetic EMPs on etched enamel surface produced prismatic structures, and showed similar mechanical properties to natural enamel. The results of X-ray diffraction (XRD) and Fourier transform infrared spectroscopy (FTIR) showed that regenerated crystal was hydroxyapatite. Molecular dynamics simulation analysis showed the binding energy between mLRAP and NAA were electrostatic forces and Van der Walls. These results introduced a promising strategy to induce crystal growth of enamel-like hydroxyapatite for biomimetic reproduction of materials with complicated hierarchical microstructures.

Odontogenesis-associated phosphoprotein truncation blocks ameloblast transition into maturation in OdaphC41*/C41* mice.

Mutations of Odontogenesis-Associated Phosphoprotein (ODAPH, OMIM *614829) cause autosomal recessive amelogenesis imperfecta, however, the function of ODAPH during amelogenesis is unknown. Here we characterized normal Odaph expression by in situ hybridization, generated Odaph truncation mice using CRISPR/Cas9 to replace the TGC codon encoding Cys41 into a TGA translation termination codon, and characterized and compared molar and incisor tooth formation in Odaph+/+, Odaph+/C41*, and OdaphC41*/C41* mice. We also searched genomes to determine when Odaph first appeared phylogenetically. We determined that tooth development in Odaph+/+ and Odaph+/C41* mice was indistinguishable in all respects, so the condition in mice is inherited in a recessive pattern, as it is in humans. Odaph is specifically expressed by ameloblasts starting with the onset of post-secretory transition and continues until mid-maturation. Based upon histological and ultrastructural analyses, we determined that the secretory stage of amelogenesis is not affected in OdaphC41*/C41* mice. The enamel layer achieves a normal shape and contour, normal thickness, and normal rod decussation. The fundamental problem in OdaphC41*/C41* mice starts during post-secretory transition, which fails to generate maturation stage ameloblasts. At the onset of what should be enamel maturation, a cyst forms that separates flattened ameloblasts from the enamel surface. The maturation stage fails completely.

Qualitative and quantitative assessment of the potential effect of cigarette smoking on enamel of human smokers' teeth.

OBJECTIVE: This study aimed to examine the potential changes in enamel surface of human smokers' teeth. MATERIALS AND METHODS: Forty extracted permanent, human, noncarious anterior teeth were used in this study. Half of these teeth were obtained from heavy smokers, while the other half of teeth were collected from nonsmokers (control teeth). The teeth were then subjected for scanning electron microscopic examination together with energy dispersive X ray and micro-hardness analysis to evaluate the qualitative and quantitative effect of smoking respectively. RESULTS: SEM of smokers' teeth showed variable degrees of destruction from small areas of demineralization as holes and pits to destruction and deterioration of the organizational pattern of the rod substance. Moreover, areas of defective remineralization were detected. The microhardness, calcium and phosphorus weight % significantly decreased whereas the Ca/P ratio was significantly increased. CONCLUSION: Cigarette smoking adversely affected the ultrastructure and mechanical properties of enamel and even hindered the normal remineralization process thus cigarette smoking cessation should be promoted in the dental office daily practices.

Gingival epithelium attachment to well- or partially cured resin composites.

Ideal restoration material for caries would allow attachment of gingival epithelia. The attachment of epithelial cells to specimens of the 4 most commercially used well- or partially-cured resin composites, with and without TEGDMA, was assessed. Effects of resin composite on the Ca9-22 gingival epithelial cell-line were assessed by measuring the cytotoxicity, viability and gene expression for attachment, apoptosis, ROS-production, pro-inflammatory cytokines, and matrix metalloproteinases. As controls, cells on tissue culture plastic or bovine tooth enamel specimens were used. Significantly less cell attachment was measured on freshly made resin-composite specimens. Concomitantly, significantly higher cytotoxicity was measured in the presence of freshly made resin-composite specimens. However, after 8 d of leakage, the cell attachment to and cytotoxicity of the resin composite was comparable to bovine tooth enamel. Significantly higher expressions of IL6, MMP2, BCL6 and ITGA4 were measured in cells attached to resin-composite surfaces than controls. There were no significant differences between the results using different conditions of resin composite, with or without TEGDMA and well or partially cured. Less cell attachment and presence of more inflammatory markers were observed on all freshly-made resin-composite surfaces. However, after a leakage period attachment of cells to the resin composite improved to the level of natural tooth materials such as enamel. This indicated that the negative effects of resin composites on epithelial cells might be transient.

Combined effects of a topical fluoride treatment and 445 nm laser irradiation of enamel against a demineralization challenge: A light and electron microscopic ex vivo study.

This study investigated the caries-preventive effect of 445 nm laser radiation in combination with fluoride on the prevention of white spot lesions. Previously, several studies have indicated the ability of 488 nm argon ion laser irradiation to reduce early enamel demineralization. A diode laser (445 nm) could be an alternative technology for possible caries-preventive potential. Each sample of a group of seventeen caries-free bovine teeth was treated in four different ways on four different zones of the labial surface: control/no treatment (C), laser irradiation only (L) (0.3 W, 60 s and applied dose of 90 J/cm2), amine fluoride application only (10,000 ppm and pH 3.9) (F), and amine fluoride application followed by laser irradiation (FL). After treatment, the teeth were subjected to a demineralization solution (pH 4.3 for 48 h at 37 °C) to induce subsurface lesions. After sectioning, the teeth were examined by light microscopy. Three teeth were analyzed by scanning electron microscopy (SEM). The depths of the subsurface lesions in the C, L, F, and FL groups were 103.01 (± 13.04), 96.99 (± 14.51), 42.59 (± 17.13), and 24.35 (± 11.38) µm, respectively. The pairwise group comparison showed the following results: p < 0.001 for FL versus C, FL versus L, F versus C, and F versus L, p = 0.019 for FL versus F and p = 0.930 for L versus C. The SEM micrographs support the light-microscopic examination. The results of the current study have shown that using relatively low irradiation settings of 445 nm laser on fluoridated enamel may be effective for prevention of white spot lesions.

Demineralization of tooth enamel following radiation therapy; An in vitro microstructure and microhardness analysis.

OBJECTIVE: The objective of this study is to evaluate the effects of radiotherapy doses on mineral density and percentage mineral volume of human permanent tooth enamel. MATERIALS AND METHODS: Synchrotron radiation Xray microcomputed tomography (SRµCT) and microhardness testing were carried out on 8 and 20 tooth samples, respectively. Enamel mineral density was derived from SRµCT technique using ImageJ software. Microhardness samples were subjected to Vickers indentations followed by calculation of microhardness and percentage mineral volume values using respective mathematical measures. Data were analyzed using paired t-test at a significance level of 5%. Qualitative analysis of the enamel microstructure was done with two-dimensional projection images and scanned electron micrographs using µCT and field emission scanning electron microscopy, respectively. RESULTS: Vickers microhardness and SRµCT techniques showed a decrease in microhardness and an increase in mineral density, respectively, in postirradiated samples. These changes were related to mineral density variation and alteration of hydroxyapatite crystal lattice in enamel surface. Enamel microstructure showed key features such as microporosities and loss of smooth homogeneous surface. These indicate tribological loss and delamination of enamel which might lead to radiation caries. CONCLUSIONS: Tooth surface loss might be a major contributing factor for radiation caries in head-and-neck cancer patients prescribed to radiotherapy. Such direct effects of radiotherapy cause enamel abrasion, delamination, and damage to the dentinoenamel junction. Suitable measures should, therefore, be worked out to protect nontarget oral tissues such as teeth while delivering effective dosages to target regions.

The effect of CO2 9.3 µm short-pulsed laser irradiation in enamel erosion reduction with and without fluoride applications-a randomized, controlled in vitro study.

The aim of this in vitro study was to evaluate the protective effect of short-pulsed CO2 9.3 µm laser irradiation against erosion in human enamel without and combined with TiF4 and AmF/NaF/SnCl2 applications, respectively, as well as compared to the protective effect of these fluoride treatments alone. After polishing, ninety enamel samples (3 × 3mm) were used for 9 different treatment groups: 4% TiF4 gel (pH 1.5, 24,533 ppm F-); AmF/NaF/SnCl2 rinse (pH 4.5; 500 ppm F-, 800 ppm Sn2); CO2 laser (average power 0.58 W); CO2 laser (0.58 W) + TiF4; CO2 laser (0.58 W) + AmF/NaF/SnCl2; CO2 laser (0.69 W); CO2 laser (0.69 W) + TiF4; CO2 laser (0.69 W) + AmF/NaF/SnCl2; negative control (deionized water). TiF4 gel was brushed on only once before the first erosive cycling, while samples treated with AmF/NaF/SnCl2 were daily immersed in 5 ml of the solution before cycling. Laser treatment occurred with a CO2 laser (wavelength 9.3 µm, pulse repetition rate 100 Hz, pulse duration 14.6 µs/18 µs, average power 0.58 W/0.69 W, fluence 1.9 J/cm2/2.2 J/cm2, beam diameter 0.63 mm, irradiation time 10 s, air cooling). TiF4 was applied only once, while AmF/NaF/SnCl2 was applied once daily before the erosive challenge. Surface loss (in µm) was measured with optical profilometry immediately after treatment, and after 5 and 10 days of erosive cycling (0.5% citric acid, pH 2.3, 6 × 2 min/day). Additionally, scanning electron microscopy investigations were performed. All application measures resulted in loss of surface height immediately after treatment. After 5 days, significantly reduced surface loss was observed after applying laser irradiation (both power settings) followed by applications of TiF4 or AmF/NaF/SnCl2 solution (p < 0.05; 2-way ANOVA and Tukey test) compared to fluoride application alone. After 10 days, compared to after 5 days, a reduced tissue loss was observed in all groups treated with AmF/NaF/SnCl2 solution. This tissue gain occurred with the AmF/NaF/SnCl2 application alone and was significantly higher when the application was combined with the laser use (p < 0.05). Short-pulsed CO2 9.3 µm laser irradiation followed by additional application of AmF/NaF/SnCl2 solution significantly reduces the progression of dental enamel erosion in vitro.

Assessment of remineralization potential of Theobromine and Sodium Fluoride gels on Artificial Caries like lesions / Avaliação do potencial de remineralização de géis de teobromina e fluoreto de sódio em lesões de cárie artificial

Objective: To compare the remineralization potential of theobromine and sodium fluoride gels on artificial caries like lesion. Materials and Methods: Forty longitudinal halves of human mandibular premolars were equally divided into 4 groups of 10 samples each: control group (C), samples were stored in distilled water during the experiment period. The remaining 30 specimens were subjected to demineralization protocol to create caries like lesions. samples were immersed for three days in a demineralization solution (pH 5.0) containing 0.2% carbopol and 0.1% lactic acid saturated with calcium phosphate tribasic. The samples were subdivided into 3 equal groups according to the treatment applied during the pH cycle. Demineralization group "D": no treatment applied. Group "F" treated with 2000 mg/liter sodium fluoride gel. Group "T" treated with 200 mg/liter theobromine gel. The specimens of the two studies groups were subjected to Demineralization- Remineralization PH Cycle Protocol for 5 days (Alternating four steps: 1: Treatment material, fluoride or theobromine ˜= 3 minutes. 2: Demineralizing solution 3 hours. 3: treatment material ˜= 3 minutes. 4: Remineralizing solution till the next cycle). The samples were investigated by scanning electron microscope (SEM) and energy dispersive x-ray analysis (EDXA). Results: The enamel of the demineralization group was porous with artificial caries like changes exposing the subsurface enamel rods with severe rod core defects. Theobromine gel and fluoride gel groups showed marked improvement in the surface characteristics in the enamel in both groups. Theobromine gel group showed more observable enamel surface improvement than the fluoride gel group. EDXA revealed that the calcium-phosphorus ratio displayed a descending order: (C > T > F > D). The differences between the two tested groups were not statistically significant. Conclusion: Theobromine gel had more effective remineralizing potential than fluoride gel as a result of its effect in improving the enamel surface characteristics of human teeth. (AU)

Objetivo: Comparar o potencial de remineralização dos géis de teobromina e fluoreto de sódio em lesões de cáries artificiais. Materiais e Métodos: Quarenta metades longitudinais de pré-molares inferiores humanos foram igualmente divididas em 4 grupos de 10 amostras cada: grupo controle (C), as amostras foram armazenadas em água destilada durante o período do experimento. As 30 amostras restantes foram submetidas ao protocolo de desmineralização paracriar lesões artificiais de cárie. As amostras foram imersas por três dias em uma solução de desmineralização (pH 5,0) contendo 0,2% de carbopol e 0,1% de ácido lático saturado com fosfato de cálcio tribásico. As amostras foram subdivididas em 3 grupos iguais, de acordo com o tratamento aplicado durante o ciclo do pH. Grupo de desmineralização "D": nenhum tratamento aplicado. Grupo "F" tratado com 2000 mg / litro de fluoreto de sódio em gel. Grupo & quot;T &q uot; tratado com 200 mg / litro de gel de teobromina. As amostras dos dois grupos de estudo foram submetidas ao protocolo de ciclo de desmineralização - remineralização por 5 dias (quatro etapas alternativas: 1: material de tratamento, flúor ou teobromina ˜= 3 minutos. 2: solução desmineralizante 3 horas. 3: material de tratamento ˜= 3 minutos 4: Solução de remineralização até o próximo ciclo). As amostras foram investigadas por microscopia eletrônica de varredura(MEV) e análise de raios-x dispersivos de energia (EDXA). Resultados: O esmalte do grupo de desmineralização era poroso, com cáries artificiais, como alterações que expunham as hastes de esmalte do subsolo com graves defeitos no núcleo da haste. Os grupos gel de teobromina e flúor apresentaram melhora acentuada nas características da superfície do esmalte nos dois grupos. O grupo gel de teobromina mostrou uma melhoria na superfície do esmalte mais observável do que o grupo gel de fluoreto. A EDXA revelou que a razão cálcio-fósforo exibia uma ordem decrescente: (C> T> F> D). As diferenças entre os dois grupos testados não foram estatisticamente significantes. Conclusão: O gel de teobromina teve um potencial remineralizante mais eficaz que o gel de flúor como resultado de seu efeito na melhoria das características da superfície do esmalte dos dentes humanos. (AU)

Artificial Caries Resistance in Enamel after Topical Fluoride Treatment and 445 nm Laser Irradiation.

OBJECTIVE: This in vitro study is aimed at investigating the caries preventive effectiveness of 445 nm diode laser in combination with topical fluoridation. MATERIALS AND METHODS: A total of 30 caries-free bovine teeth were used in this study. Eighteen teeth were covered with nail varnish except four windows on the labial surface. The windows were assigned to no treatment/control (C), laser (L) (0.3 W, 60 s, and 90 J/cm2), fluoride (F), and fluoride followed by laser (FL) treatment groups. Artificial caries lesions were created, and the teeth were sectioned and investigated under polarized light microscopy for quantitative measurement of the resulted lesion depth. Ten teeth were used for surface temperature measurement and two teeth for scanning electron microscopy (SEM). Extra twelve human molars were used for the intrapulpal temperature measurement. The absorbance of fluoride at 445 nm was measured. RESULTS: The means of lesion depth for the C, L, F, and FL groups were 123.48 (±21.93), 112.33 (±20.42), 99.58 (±30.68), and 89.03 (±30.38) µm, respectively. The pairwise differences of the L, F, and FL groups compared with the C group were significant (p < 0.05). The differences between groups were tested: FL versus L p=0.02, F versus L p=0.16, and FL versus F p=0.91, and the difference of the F versus FL was not significant (p=0.91). Temperature increment at the enamel surface and pulp roof were ∆T = 16.67 (±4.11) and 2.12 (±0.66)°C, respectively. The topical fluoride absorbance at 445 nm is five orders higher than that at 810 nm. SEM shows that after laser irradiation the enamel surface was intact and without thermal damage. CONCLUSIONS: The 445 nm laser irradiation may be useful for caries prevention, and its effectiveness is lower than those previously achieved using the argon ion laser.

The Nfic-osterix pathway regulates ameloblast differentiation and enamel formation.

Enamel makes up the outermost layer of the crown and its hardness protects other dental tissues from various stimuli. Enamel cannot be regenerated once damaged because ameloblasts are lost during the tooth eruption. Since the ameloblast differentiation mechanism is still unknown, further research is essential for developing treatments for defective or damaged enamel. Previously, we have reported that osteoblast differentiation and bone formation were regulated through the runt-related transcription factor 2 (Runx2)-nuclear factor 1-C (Nfic)-osterix (Osx) pathway where Nfic directly controls Osx expression. This pathway regulates odontoblast differentiation and dentin formation as well. The aim of this study was to investigate if the same pathway is applicable for ameloblast differentiation. Structural enamel defects with disorganized ameloblasts and decreased proliferation activity of the cervical loop were observed in Nfic-/- mice incisors. Expression of the ameloblast differentiation markers was also downregulated significantly in Nfic-/- mice. Real-time PCR analyses suggested that Runx2, Nfic, and Osx regulate the expression of ameloblast differentiation markers, where Runx2 is upstream of Nfic, and Nfic controls Osx expression. Therefore, we suggest the Runx2-Nfic-Osx pathway as one of the key factors that regulate ameloblast differentiation.

[An experimental study of the effect of extraction and retention of canine primary intruded teeth to the permanent successors].

Objective: To compare the influence of extraction and retention of the intruded dogs' teeth on permanent successors. Methods: Nine healthy 45-days-old Chinese rural puppies were selected, and six were submitted to the intrusion of the bilateral canine. Intruded teeth on the left side were extracted 30 minutes later and the teeth on the right side were kept in their sockets. After 8 months, all dogs were sacrificed. General observation, periapical radiograph and cone beam CT were used to observe the preoperative and postoperative deciduous teeth, permanent germs and permanent teeth development. The structure and content of successors' enamel were observed by scanning electron microscopy (SEM) and energy dispersive spectroscopy (EDS). Results: In the extraction groups, the enamel hypoplasia was found in 19 permanent successors, ectopic eruption in 2 cases and abnormal teeth were in 19 cases in total (79%). In the retention groups, enamel hypoplasia of the permanent successors occurred in 2 cases, root dilaceration in 6 cases, and ectopic eruption in 5 cases, retained deciduous teeth in 3 cases, and there were 22 abnormal teeth in total (92%). In blank control group, there was no abnormal teeth. The major effect of intruded deciduous teeth on the permanent successors exhibited enamel hypoplasia [85% (41/48)], presented as enamel coloration and enamel defect (P=0.416). Conclusions: The intruded deciduous teeth should be removed early in order to reduce the effect on the permanent teeth.

Effect of 2.94 µm Er: YAG laser on the chemical composition of hard tissues.

The aim was to investigate the effect of the Er-YAG laser radiation on morphology and chemical composition of enamel, dentin, and bone. The specimens of the three groups were irradiated with a very long pulse mode (VLP) of 2.94 µm Er-YAG laser with 100 mJ pulse energy and energy density of 8.42 J/ c m 2 for 30 s, at a repetition rate of 15 Hz. The organic and inorganic content of the samples were investigated by Fourier Transforms Infrared spectroscopy (FTIR). The morphological characteristics were investigated with scanning electron microscopy (SEM) and elemental analysis (calcium and phosphorus) with energy-dispersive X-ray spectroscopy (EDX). FTIR data were analyzed with a One-Way ANCOVA test and EDX data with the independent sample t-test. Following the laser radiation, FTIR showed a significant decrease in the organic content of all tissues. The weight percentage (wt %) calcium content of dentin and bone increased significantly following irradiation with a p-value of .002 for both tissues, but the wt % of phosphorus content was not influenced significantly. The morphological alterations expressed signs of fusion in all the samples.

The Role of Epithelial Stat3 in Amelogenesis during Mouse Incisor Renewal.

The aim of this study was to evaluate the role of epithelial signal transducer and activator of transcription 3 (STAT3) in mouse incisor amelogenesis. Since Stat3 is expressed in the epithelial component of developing and adult mouse teeth, we generated and analyzed Krt14Cre/+;Stat3fl/fl mutant mice in which Stat3 was inactivated in epithelia including ameloblast progenitors and ameloblasts, the cells responsible for enamel formation. Histological analysis showed little enamel matrix in mutant incisors compared to controls. Delayed incisor enamel mineralization was demonstrated using micro-computed X-ray tomography analysis and was supported by an increase in the pre-expression distance of enamel-enriched proteins such as amelogenin, ameloblastin, and kallikrein-4. Lastly, scanning electron microscopy analysis showed little enamel mineralization in mutant incisors underneath the mesial root of the 1st molar; however, the micro-architecture of enamel mineralization was similar in the erupted portion of control and mutant incisors. Taken together, our findings demonstrate for the first time that the absence of epithelial Stat3 in mice leads to delayed incisor amelogenesis.

Effect of the carbon dioxide 10,600-nm laser and topical fluoride gel application on enamel microstructure and microhardness after acid challenge: an in vitro study.

The aim of this in-vitro study was to evaluate positive effects of the carbon dioxide laser (CO2, 10,600 nm) with acidulated phosphate fluoride (APF) gel on enamel acid resistance. Twenty extracted human third molars (40 surfaces) were randomly assigned into four groups: group C, untreated control; group L, CO2 laser alone group; group F, APF 1.23% fluoride gel; and group FL, APF 1.23% gel and laser. Samples from group L were irradiated with a CO2 laser for 30s. The parameter settings used were average power, 0.73 W; time on, 100 µs; time off, 40 ms; tip-to-tissue distance, 20 mm; tip diameter 700 µm; and energy density with movements, 5 J/cm2. Samples from group F were treated with the APF gel for 4 min, and the gel was washed off with distilled water. The enamel samples from group FL were treated with APF gel for 4 min and then irradiated with the CO2 laser for 30s without removing the gel. Each enamel sample was placed in 50 ml soft drink (pH = 2.75) for 10 min then rinsed with deionized water and stored in artificial saliva at 37 °C for 1 h. Samples were assessed for Vickers hardness number (VHN) before and after treatments and subjected to SEM analysis. Data were analyzed using a one-way analysis of variance (ANOVA) and Tukey's test (α < 0.05). After the acid challenge, the untreated C group was demineralized to a great extent and the enamel surface was with the lowest mean score of microhardness. The observed VHN in the control (C group) had a mean value of 176.13, the scores in the CO2 laser group (L group) were with mean value of 238.40, the F group with a mean value of 218.45, and the fluoride-treated and laser-irradiated FL group-with a mean of 268.28 VHN. Paired t test performed to compare groups C, L, F, and FL has shown that group FL has greater resistance to decrease in microhardness of dental enamel (P ≤ 0.05) on exposure to acidic protocol. After the acid challenge, the fluoride-treated and laser-irradiated samples (group FL) showed the least diminution in enamel surface microhardness. The sub-ablative carbon dioxide laser irradiation in combination with fluoride treatment is more effective in protecting enamel surface and resisting demineralization than CO2 laser irradiation or fluoride alone.

Oral astringent stimuli alter the enamel pellicle's ultrastructure as revealed by electron microscopy.

OBJECTIVES: This electron microscopic study aimed at investigating effects of oral astringent stimuli on the enamel pellicle's morphology. METHODS: Pellicles were formed in situ within 30min on bovine enamel slabs, fixed to individuals' upper jaw splints. The pellicle-coated specimens were immersed in vitro in seven diverse astringent solutions and subsequently analyzed by scanning electron microscopy (SEM), energy dispersive X-ray (EDX) spectroscopy, as well as transmission electron microscopy (TEM). Four biocompatible astringents, namely the polyphenol epigallocatechin gallate, the metal salt iron(III) sulfate, the basic protein lysozyme, and the aminopolysaccharide chitosan, were additionally applied in situ. After rinsing the oral cavity with these compounds, the pellicle's ultrastructure was imaged by SEM and TEM, respectively. Untreated pellicle samples served as controls. RESULTS: Exposure to polyphenols and lysozyme induced particularly thicker and electron-denser pellicles in comparison to the control pellicle with similar characteristics in vitro and in situ. In contrast, acidic chitosan and metal salt solutions, respectively, revealed minor pellicle alterations. The incorporation of Fe and Al into the pellicles treated with the corresponding inorganic salts was verified by EDX analysis. CONCLUSIONS: Astringent-induced pellicle modifications were for the first time visualized by TEM. The ultrastructural alterations of the dental pellicle may partly explain the tooth-roughening effect caused by oral astringent stimuli. CLINICAL SIGNIFICANCE: Astringents might modify the pellicle's protective properties against dental erosion, attrition, as well as bacterial adhesion, and by this means may influence tooth health. The findings may thus be particularly relevant for preventive dentistry.

Salivary acquired pellicle-inspired DpSpSEEKC peptide for the restoration of demineralized tooth enamel.

Salivary acquired pellicle (SAP) is a layer of proteins and glycoproteins of salivary origin that tightly coat the tooth surface. Statherin is an important part of the SAP. The initial six-peptide sequence DpSpSEEK (where pS denotes phosphorylated serine) of the N-terminus of statherin can be immobilized on a hydroxyapatite (HAP) surface and the negatively charged domains of the DpSpSEEK side chain can catch free Ca2+ in saliva due to the charge adsorption effect. In order to prepare more functional materials based on DpSpSEEK, we designed a cysteine-labeled peptide sequence DpSpSEEKC, which could conjugate other macromolecules by forming a sulfur-based linkage. In this work, we measured the adsorption of DpSpSEEKC to HAP by various methods. We also coated DpSpSEEKC on a demineralized tooth enamel surface to evaluate its biomineralization capacity. The DpSpSEEKC-coated samples were characterized after immersion in artificial saliva for 2 weeks. The results showed that DpSpSEEKC has a strong adsorption capacity to HAP and could induce remineralization on the demineralized tooth enamel surface due to its carboxyl and phosphate groups. Compared with the control samples, the mechanical properties of the DpSpSEEKC-coated samples were obviously improved. In conclusion, DpSpSEEKC can provide a potential method for restoring demineralized tooth enamel.

Effect of Nd:YAG and CO2 Laser Irradiation on Prevention of Enamel Demineralization in Orthodontics: In Vitro Study.

OBJECTIVE: The aim of this study was to investigate Nd:YAG and CO2 laser effects in the prevention of demineralization in deeper layers of enamel via successive acid challenge cycles. BACKGROUND DATA: Lasers are promising in the prevention of enamel demineralization around the orthodontic brackets; however, there are very few studies that evaluate if the effects of treatment could be extended after successive acid challenge cycles due to permanent enamel structural alterations. MATERIALS AND METHODS: Human enamel samples were divided into five groups (n = 12): G1-application of 1.23% acidulated fluoride phosphate gel (AFP, control); G2-Nd:YAG laser irradiation (0.6 W, 84.9 J/cm2, 10 Hz, 110 µs, contact mode); G3-Nd:YAG laser irradiation associated with AFP; G4-CO2 laser irradiation (0.5 W, 28.6 J/cm2, 50 Hz, 5 µs, and 10 mm focal distance); and G5-CO2 laser irradiation associated with AFP. The samples were submitted to successive acid challenge cycles. Quantitative light-induced fluorescence and scanning electron microscopy were used to assess enamel demineralization. The data were statistically compared (α = 5%). RESULTS: G1: 50.87 ± 4.57; G2: 47.72 ± 2.87; G3: 50.96 ± 4.01; G4: 28.21 ± 2.19; and G5: 30.13 ± 6.38. The CO2 laser groups had significantly lower mineral losses than those observed in all other groups after successive acid challenge cycles. CONCLUSIONS: Only the CO2 laser (10.6 µm) irradiation prevents enamel demineralization around the orthodontic brackets even after exposure to successive acid challenges. The CO2 laser at 10.6 µm showed a deeper effect in enamel regarding caries prevention.