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Within the complex world of disaster victim identification, or DVI, forensic science practitioners use a variety of investigative techniques to work toward a common goal: identification of the decedents, bringing closure to the affected communities. Identification is a complex undertaking; the event (disaster) also can be extraordinarily complex, as it may be an acute event, or one that spans months or years. Compounding this time issue, remains may be heavily fragmented, dispersed, commingled, or otherwise disrupted by either the perpetrators or the disaster itself. To help solve these complexities, we explore the use of stable isotope analysis (SIA) in DVI events. SIA can be used with a variety of body tissues (hair, nail, bone, and teeth), and each represents different time depths in a decedent's life. Bone collagen and tooth enamel carbonate are useful to reconstruct an individual's diet and source water intakes, respectively, leading to likely population or geographic origin determinations. Additionally, the carbon and nitrogen isotopic signatures of bone collagen have calculated intraperson ranges. These facts allow investigators to determine likely origin of remains using isotopic data and can be used to link skeletal elements (to an individual), or perhaps more importantly, show that remains are not linked. Application of SIA can thus speed remains identification by eliminating individuals from short lists for identification, linking or decoupling remains, and reducing the need for some DNA testing. These strategies and hypothesis tests should commence early in the DVI process to achieve maximum effectiveness., (© 2024 American Academy of Forensic Sciences.)

Dental enamel is a mineralized extracellular matrix, and enamel defect is a common oral disease. However, the self-repair capacity of enamel is limited due to the absence of cellular components and organic matter. Efficacy of biomimetic enamel mineralization using calcium phosphate ion clusters (CPICs), is an effective method to compensate for the limited self-healing ability of fully developed enamel. Preparing and stabilizing CPICs presents a significant challenge, as the addition of certain stabilizers can diminish the mechanical properties or biosafety of mineralized enamel. To efficiently and safely repair enamel damage, this study quickly prepared CPICs without stabilizers using the atomization method. The formed CPICs were evenly distributed on the enamel surface, prompting directional growth and transformation of hydroxyapatite (HA) crystals. The study revealed that the mended enamel displayed comparable morphology, chemical composition, hardness, and mechanical properties to those of the original enamel. The approach of repairing dental enamel by utilizing ultrasonic nebulization of CPICs is highly efficient and safe, therefore indicating great promise., (© 2024 Wiley Periodicals LLC.)

Purpose: Cold atmospheric plasma can functionalize enamel without damaging the substrate morphology. It therefore has the potential to be a gentle alternative to conventional acid etching. To realize the full potential of cold atmospheric plasma in orthodontic bonding, the input parameters and protocols that are most beneficial to surface modification must first be identified. We aimed to clarify how the admixture of oxygen to cold atmospheric plasma and the rewetting of the enamel affect the conditioning properties for orthodontic bonding., Methods: First, we illustrated the morphological effects of different plasma compositions on the enamel surface by means of scanning electron microscopy. Then, we measured the shear bond strength resulting from different conditioning techniques on bovine enamel specimens: conventional acid etching; no conditioning; pure argon plasma; argon plasma plus 0.5% oxygen admixture; argon plasma plus 0.5% oxygen and rewetting after plasma application. Brackets were bonded using light cured adhesive; all specimens were subjected to thermocycling. The shear bond strength of each specimen was measured in a universal testing machine and compared using Welch one-way analysis of variance (ANOVA) and Games-Howell post hoc test., Results: Specimens conditioned with argon plasma plus 0.5% oxygen and rewetting showed a significantly higher shear bond strength than specimens conditioned with conventional acid etching. Conditioning with pure argon plasma and argon plasma plus 0.5% oxygen without rewetting yielded significantly lower shear bond strength., Conclusion: Admixing 0.5% oxygen and rewetting the enamel after plasma application are crucial steps that could help make cold atmospheric plasma a gentle conditioning technique in orthodontic bonding., (© 2023. The Author(s).)

Electron spin resonance coupled with uranium-series dating (ESR/U-series) of carbonate hydroxyapatite in tooth enamel is the main technique used to obtain age determinations from Pleistocene fossils beyond the range of radiocarbon dating. This chronological information allows to better understand diachronic change in the palaeontological record, especially with regard to the evolution of the genus Homo. Given the relative paucity of human teeth at palaeontological and archaeological localities, ESR/U-series is widely applied to the teeth of ungulate species. However, the accuracy of ESR/U-series ages is greatly affected by the incorporation of uranium in the enamel during burial in sediments. It has been shown that uranium content is positively correlated with an increased degree of atomic order in carbonate hydroxyapatite crystals, the latter determined using infrared spectroscopy. Here we present a reference infrared spectral library of tooth enamel from African ungulates, based on the grinding curve method, which serves as baseline to track the diagenetic history of carbonate hydroxyapatite in different species and thus select the best-preserved specimens for dating., (© 2024. The Author(s).)

Dental caries, the most prevalent chronic disease across all age groups, has a high prevalence, particularly among children. However, there is no specific and effective treatment for the prevention of caries in primary teeth (Pr.T.), which stems from a lack of knowledge regarding the basic nature of the tooth surface. Herein, we observed that the adhesion energies of the caries-related bacteria Streptococcus mutans and Streptococcus sanguinis to Pr.T were approximately 10 and 5.5 times higher than those to permanent teeth (Pe.T). A lower degree of mineralization and more hydrophilic characteristics of the Pr.T enamel account for this discrepancy. Accordingly, we proposed that the on-target modification of both hydroxyapatite and organic components on Pr.T by dual modification would render a sufficient hydration layer. This resulted in an approximately 11-time decrease in bacterial adhesion energy after treatment. In contrast, a single hydroxyapatite modification on Pe.T and young permanent teeth (Y.Pe.T) was sufficient to achieve a similar effect. Theoretical simulation further verified the rationality of the approach. Our findings may help understand the reason for Pr.T being caries-prone and provide references for treatment using resin restorations. This strategy offers valuable insights into daily oral hygiene and dental prophylactic treatment in children.

Identification of the sex of modern, fossil and archaeological animal remains offers many insights into their demography, mortality profiles and domestication pathways. However, due to many-factors, sex determination of osteological remains is often problematic. To overcome this, we have developed an innovative protocol to determine an animal's sex from tooth enamel, by applying label-free quantification (LFQ) of two unique AmelY peptides 'LRYPYP' (AmelY;[M+2] 2 + 404.7212 m/z) and 'LRYPYPSY' (AmelY;[M+2] 2 + 529.7689 m/z) that are only present in the enamel of males. We applied this method to eight modern cattle (Bos taurus) of known sex, and correctly assigned them to sex. We then applied the same protocol to twelve archaeological Bos teeth from the Neolithic site of Beisamoun, Israel (8-th-7-th millennium BC) and determined the sex of the archaeological samples. Since teeth are usually better preserved than bones, this innovative protocol has potential to facilitate sex determination in ancient and modern bovine remains that currently cannot be sexed., (© 2024. The Author(s).)

Objective: To evaluate the effect of four different photoactivation protocols (according to "photoactivated faces" - mesial/distal, cervical/incisal or center - and "photoactivation time" - 6-3 s) of a high-power photo activator (Valo Cordless ® -Ultradent) on the shear bond strength (SBS) between metal brackets and dental enamel and on the degree of conversion (DC) of an orthodontic resin., Materials and Methods: 40 bovine incisor crowns were randomly assigned to 4 groups (n = 10). The brackets were bonded with Transbond XT ® resin using 4 protocols according to the "photoactivation protocol" factor (which was subdivided into photoactivated faces and photoactivation time): V3C = 3 s + center; V6C = 6 s + center; V3M3D = 3 s on mesial + 3 s on distal; V3C3I = 3 s on cervical + 3 s on incisal. All the samples were stored for 4 months (water,37ºC) and then subjected to a SBS test (100KgF,1 mm/min). 40 resin discs were made to evaluate the monomer degree of conversion. Data from the SBS and DC were assessed by One-way ANOVA and Tukey's test (5%). Bond failures were analyzed according to the Adhesive Remnant Index (ARI) and evaluated by the Kruskal-Wallis test (5%)., Results: There was a statistically significant difference (p = 0.008) in the One-way ANOVA result for SBS values between all groups, but the protocols showed statistically similar results (p ≥ 0.05-Tukey's tests) concerning the photoactivated faces (V6C, V3M3D and V3C3I) and photoactivation time (V3C and V6C) factors individually. There was no statistically significant difference (p ≥ 0.05) in the One-way ANOVA result for DC values., Conclusion: The SBS and DC values will vary depending on the protocol applied., Clinical Relevance: It is possible to maintain the bracket fixation quality with the use of a high-power LED photo activator associated with a shorter photoactivation time. However, it is assumed that not all types of protocols that might be applied will provide quality bonding, such as V3C, V3M3D and V3C3I, which may - depending on the SBS and DC values - affect the final treatment time, due to brackets debonding, or increase of possibility of damage to dental enamel during bracket removal. Clinical studies are suggested to confirm the hypotheses of this research., (© 2024. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Objectives: The objective of this study was to assess how the application mode of a universal adhesive system affects the microshear bond strength (μSBS) of bulk-fill and conventional composite resins to enamel and dentin in primary teeth., Methods: A total of 80 caries-free primary second molars were randomly assigned to eight experimental groups (n = 10) based on the bonding substrate (enamel or dentin), the application mode of the universal adhesive system (etch and rinse [E&R], or self-etch [SE]), and the type of composite resin used (bulk-fill or conventional). After bonding the composite resin to enamel or dentin, the μSBS of the bonded composite resins was measured., Results: The mean μSBS value of bulk-fill composite resin was significantly higher than that of conventional composite resin for both enamel and dentin substrates, regardless of the application mode (p < 0.001). An interaction effect between the bonding substrate and the application mode of the adhesive system was observed, indicating a significant relationship (p < 0.001). The highest μSBS values for primary teeth enamel were achieved using the E&R mode with bulk-fill composite resin, while for dentin specimens, the SE mode with bulk-fill composite resin yielded the highest μSBS values. The μSBS of the E&R group was significantly higher than that of the SE group for enamel specimens (p < 0.001), whereas the μSBS of the SE group was significantly higher than that of the E&R group for dentin specimens (p < 0.001)., Conclusion: Bulk-fill composite resin demonstrated higher μSBS in comparison to conventional composite resin. The universal adhesive system exhibited superior performance in the SE mode compared to the E&R mode on primary dentin. Pre-etching the enamel before the application of the universal adhesive enhanced the μSBS to primary teeth enamel, highlighting the importance of selectively acid etching the enamel of primary teeth., (© 2024 The Author(s). Clinical and Experimental Dental Research published by John Wiley & Sons Ltd.)

Objectives: This study aimed to evaluate and compare the impact of cigarette smoking (CS) and heated tobacco (HT) on the alteration of color and ultrastructural characteristics of human enamel and cementum., Background: According to tobacco companies, a less harmful substitute for CS is HT products. Nevertheless, comprehensive research on the effects of HT on tooth structures has been lacking. This study aimed to evaluate and compare the impact of CS and HT on the alteration of color and ultrastructural characteristics of human enamel and cementum., Materials and Methods: Thirty intact and noncarious human maxillary premolars extracted for orthodontic treatment purposes, previously disinfected, were used in the study. The specimens were randomly separated into six groups (n = 10), as follows: Group 1: enamel without smoking exposure; Group 2: enamel exposed to CS; Group 3: enamel exposed to HT; Group 4: cementum without smoking exposure; Group 5: cementum exposed to CS; and Group 6: cementum exposed to HT. The measurement of color change was conducted using a spectrophotometer. The surface alterations and mineral composition of enamel and cementum were evaluated using scanning electron microscopy and energy-dispersive X-ray spectroscopy. ANOVA test followed by Tukey's post hoc test was used to determine significant differences between groups., Results: Results showed that CS had a more pronounced effect on enamel and cementum color changes than HT. The impact of CS and HT on color changes was more evident in cementum than in enamel. Surface morphology of enamel and cementum showed alterations in histology following exposure to both smoking types. Moreover, the mineral content experienced a significant reduction after using CS and HT. The reduction in calcium content after CS and HT exposure was similar. However, HT led to a significant decrease in the phosphorus content of enamel when compared with CS. At the same time, CS exposure in cementum resulted in a more significant reduction in Ca/P ratio than HT., Conclusions: Although HT may appear to present a lower danger to hard dental tissues than CS, it is not entirely harmless. CS results in more color changes on the enamel and cementum of teeth. Both smoking methods affected the mineral content of teeth, with CS having a significant effect on the roots, while HT significantly affected the crowns' mineral composition., (© 2024 The Author(s). Clinical and Experimental Dental Research published by John Wiley & Sons Ltd.)

Extracellular matrix proteins play crucial roles in the formation of mineralized tissues like bone and teeth via multifunctional mechanisms. In tooth enamel, ameloblastin (Ambn) is one such multifunctional extracellular matrix protein implicated in cell signaling and polarity, cell adhesion to the developing enamel matrix, and stabilization of prismatic enamel morphology. To provide a perspective for Ambn structure and function, we begin this review by describing dental enamel and enamel formation (amelogenesis) followed by a description of enamel extracellular matrix. We then summarize the established domains and motifs in Ambn protein, human amelogenesis imperfecta cases, and genetically engineered mouse models involving mutated or null Ambn. We subsequently delineate in silico, in vitro, and in vivo evidence for the amphipathic helix in Ambn as a proposed cell-matrix adhesive and then more recent in vitro evidence for the multitargeting domain as the basis for dynamic interactions of Ambn with itself, amelogenin, and membranes. The multitargeting domain facilitates tuning between Ambn-membrane interactions and self/co-assembly and supports a likely overall role for Ambn as a matricellular protein. We anticipate that this review will enhance the understanding of multifunctional matrix proteins by consolidating diverse mechanisms through which Ambn contributes to enamel extracellular matrix mineralization., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024. Published by Elsevier B.V.)

The micro/nano pores in natural mineralized tissues can, to a certain extent, affect their responses to mechanical loading but are generally ignored in existing indentation analysis. In this study, we first examined the void volume fraction of sound and caries lesion enamels through micro-computed tomography (micro-CT). A Berkovich indentation study was then carried out to characterize the effect of porous microstructure on the mechanical behavior of the human enamels. The indentation tests were also modeled using the nonlinear finite element analysis technique to simulate indentation load-displacement curves, which showed reasonable agreement with the experimental measurements. From the simulation results, the extent of densification in the plastic zone was identified and the corresponding stress and contact pressure evolutions were quantified. Further, a conventional elastic-perfectly plastic material model without considering micropores was also developed to investigate the compaction effect of the porous structure. The simulation results reveal that conventional elastic perfect-plastic constitutive models become less reliable to model the mechanical behavior of carious lesion enamel with increasing loss of mineral content as it underestimates the yield stress and plastic energy dissipation. This study divulges the importance of compaction of porous enamel structure beneath the indented area. Note that understanding the effect of porous microstructures on plastic behavior is vital as the involved inelastic deformation mechanism associated with irreversible processes, such as wear and localized microcracking, has a significant bearing on wear and fatigue behavior of enamel. STATEMENT OF SIGNIFICANCE: Based on micro-CT and nano-indentation characterization, a numerical model was developed aiming to precisely describe the deformation behavior of naturally porous enamel. Inelastic properties and energy dissipation characteristics of porous enamel were investigated in detail. This work demonstrated that the existence of micro-pores in White Spot Lesions (WSLs) contributes to mechanical stability, which can mitigate the reduction in Young's modulus and fracture toughness resulting from loss of mineral components. The knowledge gained from this study can be used to explain the mechanisms related to irreversible processes, such as contact induced cracking and wear, and strengthen understanding of the mechanical behavior of porous mineralized tissues., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Author(s). Published by Elsevier Ltd.. All rights reserved.)

The regeneration of demineralized enamel holds great significance in the treatment of dental caries. Amelogenin (Ame), an essential protein for mediating natural enamel growth, is no longer secreted after enamel has fully matured in childhood. Although biomimetic mineralization based on peptides or proteins has made significant progress, easily accessible, low-cost, biocompatible and highly effective Ame mimics are still lacking. Herein, we construct a series of amphiphilic branched polypeptides (CAMPs) by facile coupling of the Ame's C-terminal segment and poly(γ-benzyl-L-glutamate), which serves to simulate the Ame's hydrophobic N-terminal segment. Among them, CAMP 15 is the best biomimetic mineralization template with great self-assembly performance to guide the oriented crystallization of hydroxyapatite and is capable of inhibiting the adhesion of Streptococcus mutans and Staphylococcus aureus on the enamel surfaces. This work highlights the potential application of amphiphilic branched polypeptide as Ame mimics in repairing defected enamel, providing a promising strategy for prevention and treatment of dental caries.

Computed tomography (CT) and microcomputed tomography (μCT) require calibration against density phantoms scanned with specimens or during routine internal calibration for assessment of mineral concentration (MC) and density. In clinical studies involving bone, alternative calibration methods using bodily tissues and fluids ("phantomless" calibration) have been suggested. However, such tissues are seldom available in archeological and osteological research. This study investigates the potential of dental tissue as internal reference for calibration of μCT scans, facilitating the analysis of bone MC. We analyzed 70 molars from 24 extant primate species, including eight human teeth, each scanned with density phantoms for calibration. Our findings indicate that sampling specific regions of molars (lateral aspects of the mesial cusps) yields low variation in enamel and dentine MC values, averaging 1.27 g/cm 3 (±0.03) for dentine and 2.25 g/cm 3 (±0.03) for enamel. No significant differences were observed across molar types or among scanning procedures, including scanner model, resolution, and filters. An ad hoc test on 12 mandibles revealed low variance in MC between the conventional phantom and dental tissue calibration methods; all 36 measurements (low, medium, and high MC for each mandible) were within 0.05 g/cm 3 of each other -81% were within 0.03 g/cm 3 and 94% within 0.04 g/cm 3 . Based on these results, we propose a new "phantomless" calibration technique using these mean enamel and dentine MC values. The presented phantomless calibration method could aid in the assessment of bone pathology and enhance the scope of studies investigating bone structure and physical property variations in archeological, osteological, and laboratory-based research., (© 2024 The Authors. American Journal of Biological Anthropology published by Wiley Periodicals LLC.)

In temperate and subtropical regions, ancient proteins are reported to survive up to about 2 million years, far beyond the known limits of ancient DNA preservation in the same areas. Accordingly, their amino acid sequences currently represent the only source of genetic information available to pursue phylogenetic inference involving species that went extinct too long ago to be amenable for ancient DNA analysis. Here we present a complete workflow, including sample preparation, mass spectrometric data acquisition and computational analysis, to recover and interpret million-year-old dental enamel protein sequences. During sample preparation, the proteolytic digestion step, usually an integral part of conventional bottom-up proteomics, is omitted to increase the recovery of the randomly degraded peptides spontaneously generated by extensive diagenetic hydrolysis of ancient proteins over geological time. Similarly, we describe other solutions we have adopted to (1) authenticate the endogenous origin of the protein traces we identify, (2) detect and validate amino acid variation in the ancient protein sequences and (3) attempt phylogenetic inference. Sample preparation and data acquisition can be completed in 3-4 working days, while subsequent data analysis usually takes 2-5 days. The workflow described requires basic expertise in ancient biomolecules analysis, mass spectrometry-based proteomics and molecular phylogeny. Finally, we describe the limits of this approach and its potential for the reconstruction of evolutionary relationships in paleontology and paleoanthropology., (© 2024. Springer Nature Limited.)

The aim of this study was to provide insight into the role of camel's saliva in protecting enamel against erosion and to evaluate the levels of salivary sodium, potassium, calcium, and phosphate ions among 2 groups of camels, those with low-grade tooth surface loss and those with high-grade tooth surface loss. Twenty-eight healthy camels, 5 males and 23 females, aged 20 months to 10 years, were divided into 2 groups. Group I comprise camels with "low-grade tooth surface loss" and Group 2 with "high-grade tooth surface loss." Unstimulated saliva was collected by a draining method. The samples were then analyzed for sodium, potassium, calcium, and phosphate concentrations using automated chemistry analysis. Data were then statistically analyzed using the unpaired Student's t- test and Pearson correlation coefficient test. The study found the salivary calcium and phosphate concentrations were significantly lower in the "high-grade" erosion group, in comparison with the other group ( P  < .0001 and P   =  .0257, respectively). The results also revealed that the more extensive the tooth surface loss, the more carious lesions the subject had ( P   =  .023), and that caries are more common in the anterior teeth, particularly the incisors. The authors conclude that alterations in inorganic elements of saliva, particularly the calcium and phosphate ions, play a major role in the development of resistance to tooth surface loss. This work may represent a direction for future research to develop custom-made preventive supplements, which can protect camelid teeth against tooth surface loss., Competing Interests: Declaration of Conflicting InterestsThe author(s) declared no potential conflicts of interest with respect to the research, authorship, and/or publication of this article.

Objectives: To develop a protocol for forming subsurface caries lesions on bovine enamel by dual-species biofilms of Streptococcus mutans and Candida albicans in vitro., Design: Biofilms were grown on bovine enamel specimens in artificial saliva (AS) for seven days. After 24 h of formation, the AS was supplemented or not with fluoride (F) using sodium fluoride (0.005 or 0.008 ppm F), and the biofilms were exposed or not to a 20 % sucrose solution (reproducing a cariogenic challenge) once/day. On the seventh day, the biofilms were harvested and had their extracellular polysaccharides (EPS) and inorganic components analyzed. The specimens were subjected to computed X-ray microtomography analysis to determine their mineral concentration. Data were compared using two-way analyses of variance, followed by Fisher's LSD or Student-Newman-Keuls tests (p < 0.05)., Results: Biofilms exposed to the cariogenic challenge had significantly higher EPS concentrations than those not exposed, regardless of the presence of F. For biofilms grown with 0.008 ppm F, those exposed to the cariogenic challenge had lower F levels than those not exposed. For biofilms exposed to the cariogenic challenge, those grown with 0.008 ppm F had lower lesion depths and integrated mineral loss, and higher outer layers than those grown without F., Conclusions: The dual biofilm model assessed was able to create subsurface caries lesions in bovine enamel in vitro, which was influenced by the presence of F in the culture medium and exposure to sucrose., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Bolt's Farm is the name given to a series of non-hominin bearing fossil sites that have often been suggested to be some of the oldest Pliocene sites in the Cradle of Humankind, South Africa. This article reports the results of the first combined Uranium-Series and Electron Spin Resonance (US-ESR) dating of bovid teeth at Milo's Cave and Aves Cave at Bolt's Farm. Both tooth enamel fragments and tooth enamel powder ages were presented for comparison. US-ESR, EU and LU models are calculated. Overall, the powder ages are consistent with previous uranium-lead and palaeomagnetic age estimates for the Aves Cave deposit, which suggest an age between ~3.15 and 2.61 Ma and provide the first ages for Milo's Cave dates to between ~3.1 and 2.7 Ma. The final ages were not overly dependent on the models used (US-ESR, LU or EU), which all overlap within error. These ages are all consistent with the biochronological age estimate (<3.4->2.6 Ma) based on the occurrence of Stage I Metridiochoerus andrewsi . Preliminary palaeomagnetic analysis from Milo's Cave indicates a reversal takes place at the site with predominantly intermediate directions, suggesting the deposit may date to the period between ~3.03 and 3.11 Ma within error of the ESR ages. This further suggests that there are no definitive examples of palaeocave deposits at Bolt's Farm older than 3.2 Ma. This research indicates that US-ESR dating has the potential to date fossil sites in the Cradle of Humankind to over 3 Ma. However, bulk sample analysis for US-ESR dating is recommended for sites over 3 Ma., Competing Interests: The authors declare that they have no competing interests., (© 2024 Yu et al.)

Objective: This study compared three protocols for developing artificial white spot lesions (WSL) using biofilm models., Methodology: In total, 45 human enamel specimens were sterilized and allocated into three groups based on the biofilm model: Streptococcus sobrinus and Lactobacillus casei (Ss+Lc), Streptococcus sobrinus (Ss), or Streptococcus mutans (Sm). Specimens were incubated in filter-sterilized human saliva to form the acquired pellicle and then subjected to the biofilm challenge consisting of three days of incubation with bacteria (for demineralization) and one day of remineralization, which was performed once for Ss+Lc (four days total), four times for Ss (16 days total), and three times for Sm (12 days total). After WSL creation, the lesion fluorescence, depth, and chemical composition were assessed using Quantitative Light-induced Fluorescence (QLF), Polarized Light Microscopy (PLM), and Raman Spectroscopy, respectively. Statistical analysis consisted of two-way ANOVA followed by Tukey's post hoc test (α=0.05). WSL created using the Ss+Lc protocol presented statistically significant higher fluorescence loss (ΔF) and integrated fluorescence (ΔQ) in comparison to the other two protocols (p<0.001)., Results: In addition, Ss+Lc resulted in significantly deeper WSL (137.5 µm), followed by Ss (84.1 µm) and Sm (54.9 µm) (p<0.001). While high mineral content was observed in sound enamel surrounding the WSL, lesions created with the Ss+Lc protocol showed the highest demineralization level and changes in the mineral content among the three protocols., Conclusion: The biofilm model using S. sobrinus and L. casei for four days was the most appropriate and simplified protocol for developing artificial active WSL with lower fluorescence, higher demineralization, and greater depth.

Vertebrates use the phosphate mineral apatite in their skeletons, which allowed them to develop tissues such as enamel, characterized by an outstanding combination of hardness and elasticity. It has been hypothesized that the evolution of the earliest vertebrate skeletal tissues, found in the teeth of the extinct group of conodonts, was driven by adaptation to dental function. We test this hypothesis quantitatively and demonstrate that the crystallographic order increased throughout the early evolution of conodont teeth in parallel with morphological adaptation to food processing. With the c-axes of apatite crystals oriented perpendicular to the functional feeding surfaces, the strongest resistance to uniaxial compressional stress is conferred along the long axes of denticles. Our results support increasing control over biomineralization in the first skeletonized vertebrates and allow us to test models of functional morphology and material properties across conodont dental diversity., (© 2024. The Author(s).)

Objective: To evaluate the enamel bonding ability and orthodontic adhesive resin degree of conversion using the experimental bracket design. Material and Methods . Thirteen bovine teeth were used in the study. The experimental bracket was modified with a translucent region in the center of its body. After enamel etching, Orthocem orthodontic adhesive (FGM, Joinville, Brazil) was applied on the bracket base for bonding. The groups were divided as follows ( n = 10 per group): (1) control (CB) with standard brackets and (2) spot bracket (SB) with experimental brackets featuring a 0.8 mm translucent region at the center using carbide bur. Shear bond strength (SBS) was evaluated after 24 hours in a universal testing machine and adhesive remnant index (ARI). The degree of conversion (DC) was analyzed using Raman spectroscopy ( n = 3 per group). Data were then analyzed using Student's t -test and Mann-Whitney statistical methods., Results: The SB group exhibited a higher mean SBS (10.33 MPa) compared to the CB Group (8.77 MPa). However, there was no statistical difference between the groups ( p = 0.376). Both SB and CB groups had a mean ARI score of 1. Raman analysis revealed a higher degree of conversion in the SB group (49.3%) compared to the CB group (25.9%)., Conclusions: The experimental support showed a higher degree of adhesive conversion, although there was no significant increase in bond strength., Competing Interests: The authors declared no potential conflicts of interest concerning the research, authorship, and/or publication of this article., (Copyright © 2024 Alexandre Daher Yunes Salgado et al.)

Background: Remineralization of dental enamel is an important intervention strategy for the treatment of demineralized lesions. Existing approaches have limitations such as failure to adequately reproduce both the ideal structural and mechanical properties of the native tooth. The ability of ultrasound to control and accelerate the crystallization processes has been widely reported. Therefore, a new approach was explored for in-vitro enamel remineralization involving the synergistic effect of high-intensity focused ultrasound (HIFU) coupled with calcium phosphate ion clusters (CPICs)., Methods: The demineralized enamel was treated with CPICs, with or without subsequent HIFU exposure for different periods (2.5, 5, and 10 min). The specimens were characterized by scanning electron microscopy (SEM), atomic force microscopy (AFM), and Raman spectroscopy. The surface hardness and crystallographic properties of the treated specimens were evaluated using Vickers microhardness testing and X-ray diffraction (XRD), respectively., Results: SEM revealed distinct, organized, and well-defined prismatic structures, showing clear evidence of remineralization in the combined CPIC/HIFU treatment groups. AFM further revealed a decrease in the surface roughness values with increasing HIFU exposure time up to 5 min, reflecting the obliteration of interprismatic spaces created during demineralization. The characteristic Raman band at 960 cm -1 associated with the inorganic phase of enamel dominated well in the HIFU-treated specimens. Importantly, microhardness testing further demonstrated that new mineral growth also recovered the mechanical properties of the enamel in the HIFU-exposed groups. Critical to our aspirations for developing this into a clinical process, these results were achieved in only 5 min., Conclusion: HIFU exposure can synergise and significantly accelerate in-vitro enamel remineralization process via calcium phosphate ion clusters. Therefore, this synergistic approach has the potential for use in future clinical interventions., Competing Interests: Dr Barsha Shrestha reports grants from the National Health and Medical Research Council grant and grants from the Australian Dental Research Foundation grant, during the conduct of the study. Mrs Sheetal Maria Rajan reports grants from the National Health and Medical Research Council and grants from the Australian Dental Research Foundation, during the conduct of the study. Dr Sultan Aati reports grants from the National Health and Medical Research Council and grants from the Australian Dental Research Foundation, during the conduct of the study. Dr Emielda Yusiharni reports grants from the National Health and Medical Research Council grant (NMHRC, 1188401) and grants from the Australian Dental Research Foundation grant (ADRF; 0080045000900), during the conduct of the study. Dr Martin Saunders reports grants from the National Health and Medical Research Council and grants from the Australian Dental Research Foundation, during the conduct of the study. Dr. Amr Fawzy (lead chief investigator) reports grants from the National Health and Medical Research Council grant (NMHRC, 1188401) and grants from the Australian Dental Research Foundation grant (ADRF; 0080045000900), during the conduct of the study. The authors declare no other competing interests in this work., (© 2024 Shrestha et al.)

The Middle Ages in the Iberian Peninsula is a period of special interest for studying the relationship of climate change with historical and socioeconomic processes. Between the 8th and 15th centuries AD, the Peninsula was characterized not only by complex political, cultural, and social transitions but also by major variations in the climate. The objective of this study was to examine differences in diet and mobility between distinct populations of the Peninsula and explore the possible relationship of diet, mobility, and culture with environmental variables and geographical settings. For this purpose, we obtained stable isotopic ratios of carbon and oxygen (δ13C and δ18O) from the enamel apatite of first upper incisors from 145 individuals at eight archeological sites that represent both Christian and Islamic communities and both rural and urban social settings. Results revealed a dietary difference between Christian and Islamic populations, observing a greater contribution of C4 plants, possibly sorghum, in the diet of the latter, especially in a rural setting. The disparity in oxygen isotopic ratios between populations from the North and South of the Peninsula is consistent with modern climatic differences between these regions. In this line, intraregional variability in oxygen isotopic ratios may hint at diachronic occupation phases under varying climatic conditions. The few isotopic outliers in our sample suggest overall low mobility levels., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2024 Martín-Alonso et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

The remarkable damage-tolerance of enamel has been attributed to its hierarchical microstructure and the organized bands of decussated rods. A thorough characterization of the microscale rod evolution within the enamel is needed to elucidate this complex structure. While prior efforts in this area have made use of single particle tracking to track a single rod evolution to various degrees of success, such a process can be both computationally and labor intensive, limited to the evolution path of a single rod, and is therefore prone to error from potentially tracking outliers. Particle image velocimetry (PIV) is a well-established algorithm to derive field information from image sequences for processes that are time-dependent, such as fluid flows and structural deformation. In this work, we demonstrate the use of PIV in extracting the full-field microstructural distribution of rods within the enamel. Enamel samples from a wild African lion were analyzed using high-energy synchrotron X-ray micro-tomography. Results from the PIV analysis provide sufficient full-field information to reconstruct the growth of individual rods that can potentially enable rapid analysis of complex microstructures from high resolution synchrotron datasets. Such information can serve as a template for designing damage-tolerant bioinspired structures for advanced manufacturing. STATEMENT OF SIGNIFICANCE: Thorough characterization and analysis of biological microstructures (viz. dental enamel) allows us to understand the basis of their excellent mechanical properties. Prior efforts have successfully replicated these microstructures via single particle tracking, but the process is computationally and labor intensive. In this work, optical flow imaging algorithms were used to extract full-field microstructural distribution of enamel rods from synchrotron X-ray computed tomography datasets, and a field method was used to reconstruct the growth of individual rods. Such high throughput information allows for the rapid production/prototyping and advanced manufacturing of damage-tolerant bioinspired structures for specific engineering applications. Furthermore, the algorithms used herein are freely available and open source to broaden the availability of the proposed workflow to the general scientific community., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Acta Materialia Inc. All rights reserved.)

Background: Yogurt, due to its rich casein phosphopeptide (CPP) content could promote remineralization of enamel. Contrary to the age-old usage of animal milk based yogurt, vegan dairy products are gaining momentum due to multiple reasons. Considering this change, the aim of the present study was to assess the in vitro effect of animal and plant-based yogurt extracts on demineralization of enamel., Methods: Enamel windows were prepared on the crowns of sixty premolar teeth using nail paint. Teeth were divided into four groups of fifteen each and were treated with distilled water, demineralizing agent, solution with a mixture of demineralizing agent and yogurt supernatants, respectively for 96 hrs. Quantitative analysis was done using EDXRF (baseline and postexperimental calcium and phosphorus content). Additionally, confocal microscopic analysis was performed to assess the extent of demineralization., Results: The animal-based yogurt (Group III) had the highest post experimental calcium value (mean±SD=81.15±5.02) and positive percentage of change in calcium level (15%; P=0.007) among the groups. This was followed by plant-based yogurt (Group IV) (mean calcium= 76.18±5.12; positive percentage change =8.11%; P=0.003)., Conclusions: Animal based yogurt could provide higher protection against enamel demineralization when compared to plant-based yogurt.

Objective: To evaluate the effects of metal primer II (MP II) on the shear bond strength (SBS) of orthodontic brackets bonded to teeth and bis-acryl composite provisional material (Bis-Acryl)., Material and Methods: Twenty extracted human premolars specimens and 20 premolar shaped Bis-Acryl specimens were obtained and randomly divided into two surface groups. The first group consisted of human premolars (T) bonded to brackets in the conventional way while in the second (T-MP) MP II was applied on the bracket base before bonding. Similarly, one group of provisional material (PM) was prepared according to conventional treatment and another with the application of MP-II metal bonder (PM-MP). In all cases Ortho-brackets (Victory Series, 3 M) were bonded employing Transbond XT resin cement. Then the brackets were debonded under shear and the results were statistically analyzed by two-way analysis of variance and Holm Sidak at α = .05. The debonded surfaces of all specimens were examined by light microscopy and the Adhesive Remnant Index (ARI) was recorded., Results: The SBS results exhibited significant differences er (p < .001). For both the T and TM the application of MP-II increased the SBS compared to respective control groups (p < .001). The T-C group was found inferior compared to PM-C (p < .001) and the same is true for the comparison between T-MP and PM-MP (p < .001). ARI indexes demonstrated that the tooth groups were characterized by a predominantly adhesive failure at the resin-dentin interface. In contrast, the control group for provisional crowns (PM-C) showed a predominantly cohesive failure mode, which moved to predominantly adhesive after the application of MP II., Conclusion: The application of MP II enhances the SBS on both, human enamel and provisional crown materials., (© 2024 The Authors. Clinical and Experimental Dental Research published by John Wiley & Sons Ltd.)

Background: White spot lesions (WSLs) are frequently linked with low microhardness and mineral content changes. several strategies have been employed to deal with these problems. This investigation aimed to analyze the microhardness and mineral content changes after remineralization with bioactive glass (BAG) and casein phospho-peptide-amorphous calcium phosphate with fluoride (CPP-ACPF)., Methods: Twenty sound maxillary first premolars extracted were used to obtain a total of one hundred enamel samples. forty enamel slabs were split into four experimental groups (n = 10 each): Group I, BAG; Group II, BAG+CPP-ACPF; Group III, CPP-ACPF varnish; and Group IV, artificial saliva (negative control). To create artificial WSLs, all samples were preserved in a prepared demineralizing agent for 72 h before treatment with remineralizing agents. Vickers microhardness test was performed. Additionally, 60 enamel samples were selected for analysis using energy dispersive spectroscopy (EDX) and assigned to six experimental groups; the first four groups were similar to that used in the microhardness test along with Group V: WSLs, and Group VI: baseline. The statistical analyses employed in this study included Tukey's HSD (p<0.05), one-way ANOVA, and Shapiro-Wilk., Result: Regarding surface microhardness, the BAG+CPP-ACPF group showed the most favorable recovery, which was better than the outcomes of the BAG and CPP-ACPF groups. A statistically significant change (p <0.05) was not observed between them. Similarly, for mineral content change, the BAG+CPP-ACPF group demonstrated the greatest result, The BAG group came next, and the CPP-ACPF group came last., Conclusion: The BAG+CPP-ACPF group might be regarded as the best course of treatment for enhancing both the surface microhardness and mineral content (Ca, P), while the control group (Artificial saliva) showed the least satisfactory results in comparison. After demineralization, mineral content and microhardness decreased in all samples. Therefore, BAG+CPP-ACPF significantly improved the surface microhardness and mineral content., Competing Interests: No competing interests were disclosed., (Copyright: © 2024 Ghanim Rahman R and Diab BS.)

Interpretations of Late Pleistocene hominin adaptative capacities by archaeologists have focused heavily on their exploitation of certain prey and documented contemporary behaviours for these species. However, we cannot assume that animal prey-taxa ecology and ethology were the same in the past as in the present, or were constant over archaeological timescales. Sequential isotope analysis of herbivore teeth has emerged as a particularly powerful method of directly reconstructing diet, ecology and mobility patterns on sub-annual scales. Here, we apply 87 Sr/ 86 Sr isotope analysis, in combination with δ 18 O and δ 13 C isotope analysis, to sequentially sampled tooth enamel of prevalent herbivore species that populated Europe during the Last Glacial Period, including Rangifer tarandus, Equus sp. and Mammuthus primigenius. Our samples come from two open-air archaeological sites in Central Germany, Königsaue and Breitenbach, associated with Middle Palaeolithic and early Upper Palaeolithic cultures, respectively. We identify potential inter- and intra-species differences in range size and movement through time, contextualised through insights into diet and the wider environment. However, homogeneous bioavailable 87 Sr/ 86 Sr across large parts of the study region prevented the identification of specific migration routes. Finally, we discuss the possible influence of large-herbivore behaviour on hominin hunting decisions at the two sites., (© 2024. The Author(s).)

Teeth are considered unique as fingerprints for identification purposes. Their structure and resilience mean they can remain for thousands and millions of years withstanding extreme conditions, including burning. During burning, bones undergo carbonization at approximately 400°C and calcination at approximately 700°C. This study aimed to investigate the effects of carbonization and calcination on dental tissue. It involved nondestructive analyses of 58 extracted human teeth before and after burning, using x-ray diffraction, micro-CT, and high-resolution confocal microscopy. The results revealed that during carbonization, dentin volume decreased in two thirds of the sample, accompanied by crack formation and significant reduction in hydroxyapatite crystal size (p<0.001). During calcination, dentin volume decreased in all teeth, along with a significant deepening of the cracks (p<0.001), while enamel crystal size increased slightly. Initial changes in teeth occurred at lower temperatures than had once been assumed, as indicated by the cracks during carbonization, and there was up to a 36% decrease in dentin volume during calcination, which should be considered when measuring burnt teeth. The results of this research provide new insight into understanding dental tissue response to burning. Thus, dental remains may contribute to the knowledge needed to reconstruct anthropological and forensic scenarios involving burning., Competing Interests: Declaration of Competing Interest None, (Copyright © 2024 Elsevier B.V. All rights reserved.)

Biocompatible magnesium alloys represent revolutionary implantable materials in dentistry and orthopedics but face challenges due to rapid biocorrosion, necessitating protective coatings to mitigate dysfunction. Directly integrating durable protective coatings onto Mg surfaces is challenging because of intrinsic low coating compactness. Herein, inspired by tooth enamel, a novel highly compact dual-protection inorganic-protein (inorganicPro) coating is in situ constructed on Mg surfaces through bovine serum albumin (BSA) protein-boosted reaction between sodium fluoride (NaF) and Mg substrates. The association of Mg ions and BSA establishes a local hydrophobic domain that lowers the formation enthalpy of NaMgF 3 nanoparticles. This process generates finer nanoparticles that function as "bricks," facilitating denser packing, consequently reducing voidage inside coatings by over 50% and reinforcing mechanical durability. Moreover, the incorporation of BSA in and on the coatings plays two synergistic roles: 1) acting as "mortar" to seal residual cracks within coatings, thereby promoting coating compactness and tripling anticorrosion performance, and 2) mitigating fouling-accelerated biocorrosion in complex biosystems via tenfold resistance against biofoulant attachments, including biofluids, proteins, and metabolites. This innovative strategy, leveraging proteins to alter inorganic reactions, benefits the future coating design for Mg-based and other metallic materials with tailored anticorrosion and antifouling performances., (© 2024 Wiley‐VCH GmbH.)

This study aimed to assess the in vitro effects of re-irradiation on enamel and dentin properties, simulating head and neck cancer radiotherapy retreatment. Forty-five human permanent molars were classified into five groups: non-irradiated; irradiated 60 Gy, and re-irradiated with doses of 30, 40, and 50 Gy. Raman spectroscopy, scanning electron microscopy (SEM), and energy dispersive x-ray spectroscopy (EDS) were employed for analysis. Raman spectroscopy assessed intensity, spectral area, and specific peaks comparatively. Statistical analysis involved Kolmogorov-Smirnov and One-Way ANOVA tests, with Tukey's post-test (significance level set at 5%). Significant changes in irradiated, non-irradiated, and re-irradiated enamel peaks were observed, including phosphate (438 nm), hydroxyapatite (582 nm), phosphate (960 nm), and carbonate (1070 nm) (p < 0.05). Re-irradiation affected the entire tooth (p > 0.05), leading to interprismatic region degradation, enamel prism destruction, and hydroxyapatite crystal damage. Dentin exhibited tubule obliteration, crack formation, and progressive collagen fiber fragmentation. EDX revealed increased oxygen percentage and decreased phosphorus and calcium post-reirradiation. It is concluded that chemical and morphological changes in irradiated permanent teeth were dose-dependent, exacerbated by re-irradiation, causing substantial damage in enamel and dentin., (© 2024. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

The transition from hunting-gathering to agriculture stands as one of the most important dietary revolutions in human history. Yet, due to a scarcity of well-preserved human remains from Pleistocene sites, little is known about the dietary practices of pre-agricultural human groups. Here we present the isotopic evidence of pronounced plant reliance among Late Stone Age hunter-gatherers from North Africa (15,000-13,000 cal BP), predating the advent of agriculture by several millennia. Employing a comprehensive multi-isotopic approach, we conducted zinc (δ 66 Zn) and strontium ( 87 Sr/ 86 Sr) analysis on dental enamel, bulk carbon (δ 13 C) and nitrogen (δ 15 N) and sulfur (δ 34 S) isotope analysis on dentin and bone collagen, and single amino acid analysis on human and faunal remains from Taforalt (Morocco). Our results unequivocally demonstrate a substantial plant-based component in the diets of these hunter-gatherers. This distinct dietary pattern challenges the prevailing notion of high reliance on animal proteins among pre-agricultural human groups. It also raises intriguing questions surrounding the absence of agricultural development in North Africa during the early Holocene. This study underscores the importance of investigating dietary practices during the transition to agriculture and provides insights into the complexities of human subsistence strategies across different regions., (© 2024. The Author(s).)

Teeth exemplify architectures comprising an interplay of inorganic and organic constituents, resulting in sophisticated natural composites. Rodents (Rodentia) showcase extraordinary adaptations, with their continuously growing incisors surpassing human teeth in functional and structural optimizations. In this study, employing state-of-the-art direct atomic-scale imaging and nanoscale spectroscopies, we present compelling evidence that the release of material from ameloblasts and the subsequent formation of iron-rich enamel and surface layers in the constantly growing incisors of rodents are complex orchestrated processes, intricately regulated and independent of environmental factors. The synergistic fusion of three-dimensional tomography and imaging techniques of etched rodent́s enamel unveils a direct correlation between the presence of pockets infused with ferrihydrite-like material and the acid resistant properties exhibited by the iron-rich enamel, fortifying it as an efficient protective shield. Moreover, observations using optical microscopy shed light on the role of iron-rich enamel as a microstructural element that acts as a path for color transmission, although the native color remains indistinguishable from that of regular enamel, challenging the prevailing paradigms. The redefinition of "pigmented enamel" to encompass ferrihydrite-like infusion in rodent incisors reshapes our perception of incisor microstructure and color generation. The functional significance of acid-resistant iron-rich enamel and the understanding of the underlying coloration mechanism in rodent incisors have far-reaching implications for human health, development of potentially groundbreaking dental materials, and restorative dentistry. These findings enable the creation of an entirely different class of dental biomaterials with enhanced properties, inspired by the ingenious designs found in nature.

Objectives: This in vitro study evaluated the effect of different colouring solutions and primer systems used in the bonding of brackets on enamel colour change and bond strength., Materials and Methods: 120 premolar teeth were divided into four main groups; brackets were bonded with 37% orthophosphoric acid + Transbond XT Primer in Group 1, 3 M Single Bond Universal in Group 2, Transbond Plus SEP in Group 3, and G-Premio Bond in Group 4. Each group was divided into three subgroups, and the teeth were placed in a cup containing coffee and tea mixture, in a cup containing cola and in distilled water. A bond strength test was applied to all teeth. Colour measurements of all teeth were performed at 2 different times: before bonding and after the bond strength test., Results: The average bond strength of the 37% orthophosphoric acid group was higher than that of the other groups. The effect of primer and solution groups on colour change was statistically significant (p = 0.001 and p = 0.023, respectively)., Conclusions: In this study, the bond strength was clinically sufficient in all primer groups. The highest colour change was observed when the tea-coffee solution and Transbond Plus SEP primer were used., Clinical Relevance: This study has identified enamel discoloration and bond strength from different colouring solutions and primer systems used for bonding braces, which can be used to inform clinicians and patients to achieve better treatment results., (© 2024. The Author(s).)

Aims - to compare the color changes, the surface roughness and morphology of the enamel bleached with two different bleaching solutions (chemical and laser activated), preceded or not with acid etching. Thirty teeth of bovine prepared and haphazardly assigned to 2 groups (n=15) depending on bleaching technique. Each group subdivided to 3 subgroup (n=5) consistent with acid etching by 37% phosphoric acid. Atomic force microscopy and VITA easy shade spectrophotometer were performed twice for all the specimens before and after bleaching. ANOVA, the Paired sample t-test, and the independent sample t-test used for statistical analysis. As for the color changes, the groups that were bleached by the chemical method, the difference among the three subgroups was statistically significant. This also applies to the groups bleached with the laser method. When comparing the results of the chemical bleaching subgroups with the laser bleaching ones, the difference was not significant. Roughness results showed significant differences between certain subgroups and non-significant differences among others. However, the difference was statistically significant between the chemical and laser groups, laser technique resulted in less surface roughness than the chemical one. Acid etching before bleaching produced better colour change in both the chemical and laser assisted bleaching. In chemical bleaching, surface roughness was higher when acid etching was used. This was also true for laser bleaching technique. In general, laser assisted bleaching produced less surface roughness than chemical bleaching.

Objective: The present study aimed to evaluate the effect of remineralizing agents on demineralized enamel intended for use as fluoride substitutes or supplements for oral hygiene applications., Methodology: Enamel samples were obtained from 30 bovine teeth. The enamel blocks were stored in 20 mL of demineralization solution for 72 h. They were then brushed with the following toothpaste for the remineralization protocol: NaF, NaF/SnF 2 combination, NovaMin, or nano-hydroxyapatite. SEM/EDX examinations and microhardness measurements of the samples were performed to investigate the remineralization efficacy of the studied toothpaste. One-way analysis of variance (ANOVA) with post hoc Tukey's HSD test was used to analyze the change in microhardness values in different remineralization protocols (p < 0.05)., Results: Differences in the mean remineralization (%RP) and hardness recovery (%HR) were determined between the groups (p < 0.05). Groups 1 and 4 showed significant differences in %RP (p < 0.05). In the SEM/EDX examinations, the samples treated with n-HAp showed an accumulation of crystal deposits on the enamel surface, although at a lower density than those treated with NaF and NaF/SnF 2 combination., Conclusion: The remineralization strategy in toothpaste plays an important role in enamel remineralization. NovaMin-containing toothpaste showed positive effects on the enamel surface with better Ca/P ratio. Toothpastes containing n-HAp triggered less change in the increase of microhardness values compared to other toothpastes. The use of SnF 2 in toothpaste in combination with NaF significantly increased the binding of fluoride to demineralized enamel compared to toothpaste containing NaF alone., (© 2023 Wiley Periodicals LLC.)

Isotopic analysis of human tooth enamel can provide life history information useful in forensic identification. These applications depend on the availability of reference data documenting isotopic values for individuals with known life history and on the comparability of data from reference and case work samples. Here we build on previous methodological research, which has largely focused on paleontological and archaeological samples, and conduct experiments using enamel from modern human teeth targeting three sample preparation variables (sample particle size, storage conditions, and chemical pretreatments). Our results suggest that differences in particle size affect the efficiency of sample reactions during pretreatment and analysis, with coarse particles giving reduced loss of enamel carbonate during acid pretreatments but producing slightly higher oxygen isotope values than fine particles during analysis. Data for samples stored in dry and ambient environments following pretreatment were indistinguishable, suggesting no exchange of oxygen between samples and ambient water vapor. Finally, chemical pretreatments with a range of commonly used reactants and conditions showed a pervasive, moderate oxygen isotope shift associated with acetic acid treatment, which may be caused by exchange of enamel hydroxyl groups with reagents or rinse waters. Collectively, the results emphasize the importance of methodological standardization to improve comparability and reduce potential for bias in the forensic application of tooth enamel isotope data., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Chartered Society of Forensic Sciences. Published by Elsevier B.V. All rights reserved.)

Objective: The effects of four toothpastes on the color stability of in-office bleached tooth specimens were determined., Materials and Methods: We evaluated an experimental toothpaste (EXP) and three commercially available toothpastes: Colgate Optic White (OPW), Aquafresh White & Protect (AWP), and Crest 3D White (CDW). OPW, AWP, and CDW contained inorganic abrasives, whereas EXP and AWP contained sodium polyphosphate. Forty-eight randomly selected human-extracted maxillary central incisors were bleached and brushed twice daily over 30 days. We analyzed the final color difference (ΔE*ab, ΔE 00 , ΔWI D ), arithmetic average surface roughness (Ra) of the enamel measured on days 0 and 30, and scanning electron microscopy images of enamel surfaces and toothpastes. ΔE*ab, ΔE 00 , ΔWI D , and Ra were analyzed using one-way analysis of variance and Tukey's test (α = 0.05)., Results: ΔE*ab and ΔE 00 values were significantly lower after toothbrushing with EXP, OPW, and CDW than with AWP. OPW induced the greatest positive ΔWI D . Ra was significantly increased by OPW and CDW, but slightly increased by AWP, with cube-like particles, and EXP, with no particle-like structures., Conclusions: Only EXP stabilized the color of bleached teeth without increasing the enamel surface roughness. Sodium polyphosphate with approximately 10 phosphate groups was effective at removing stains., Clinical Significance: The effect of toothpaste on the color stability of bleached teeth depends on the constituting abrasives and chemical components. Polyphosphoric acid has different stain-removal effects depending on its degree of polymerization. Additionally, although certain types of abrasives may be effective for color stability, they also increase the surface roughness of the enamel., (© 2023 Wiley Periodicals LLC.)

The present work aimed at assessing chemical, topographical, and morphological changes induced by Nd : YAG laser treatment of dental enamels by means of energy dispersive X-ray spectroscopy (EDS), atomic force microscopy (AFM), and scanning electron microscopy (SEM). Fifteen human enamel specimens were obtained, three of samples were kept untreated as a control while the others twelve samples were equally divided into four groups where each group have a three samples according to treating approach as: G1:(untreated);G2: (treated with Nd:YAG laser, 100 mJ/pulse,10 Hz/1064nm); G3(treated with Nd:YAG laser, 500 mJ/pulse, 10 Hz/1064nm); G4(treated with Nd:YAG laser 1000 mJ/pulse, 10 Hz/1064nm), and finally G5(treated with Nd:YAG laser, 1000 mJ/pulse, 10 Hz/532nm) respectively. Beside many craters and cracks, the AFM results showed fractures with depths of 19.23 nm, 174.7 nm, 216.9 nm, 207.4 nm and 156.5 nm and width of 559.2 nm, 833.4 nm, 1115 nm, 695.0 nm, and 5142 nm for all Groups respectively. The highest surface roughness was found in G5 with 111.4 nm while the lowest surface roughness was found in G1 to be 14.3 nm. The inside surface of the fissures was also rough. The SEM micrographs revealed modifications to the morphology. EDS was used to measure the phosphorous (P), calcium (Ca), oxygen (O), and carbon (C) percentages presented in crater areas and their surroundings, Ca, P, O, and C levels were observed to vary significantly at the crater and its rim, a lower percentage of C wt% were realized corresponding to laser treatment of 1000 mJ/Pulse laser energy. However, it was not feasible to recognize a specific chemical arrangement in the craters. It is also concluded that the higher depth and particular edge of ablated part when teeth were irradiated by laser with 1000 mJ/10Hz/1064nm., (© 2024. The Author(s), under exclusive licence to Springer-Verlag London Ltd., part of Springer Nature.)

Introduction: The aim of this in vitro study was to assess the suitability of high-resolution time-of-flight secondary ion mass spectrometry (ToF-SIMS) for visualizing cross-sectional changes in human enamel microstructure and chemical composition during treatment and remineralization cycling of artificially generated caries lesions underneath an artificial plaque., Methods: Treatments consisted of exposure to twice daily toothpaste/water slurries prepared from 0, 1,100, and 5,000 μg/g fluoride (F) NaF/silica toothpastes. In addition, treatments with slurries prepared from 1,100 μg/g F SnF2/silica toothpastes were done using 44Ca in the remineralization solution to allow for differentiation of newly formed mineral and exploration of incorporated metal dopants using ToF-SIMS. Complementary microhardness, scanning electron microscopy, and high-resolution transmission electron microscopy (HR-TEM) investigations were performed on enamel cross sections., Results: HR-TEM was used for the first time to determine the change in crystallinity during remineralization revealing distinct microstructural zones within one lesion. Chemical mapping using ToF-SIMS demonstrated that the distribution of F, while observed primarily in the new mineral phase, was widespread throughout the lesion with 44Ca substantially limited to the remineralizing mineral. Both penetrated the inter-rod spaces of the sound enamel illustrating how acid damage propagates into the native mineral as the caries lesion deepens. HR-TEM examination revealed different regions within the lesion characterized by distinct micro- and ultrastructures. Importantly, HR-TEM revealed a return of crystallinity following remineralization. F dose-response observations verified the ability of these high-resolution techniques to differentiate remineralization efficacy., Conclusion: The collective results provided new insights such as the visualization of F or calcium penetration pathways, as well as new tools to study the caries process., (© 2024 The Author(s). Published by S. Karger AG, Basel.)

Introduction: Titanium tetrafluoride (TiF4) is an anticariogenic agent with high remineralizing potential. However, the acidic pH of TiF4 solution can limit its clinical application. The present study aimed to prepare and characterize a new TiF4-dendrimer inclusion complex and evaluate its ability to inhibit enamel demineralization under pH cycling conditions., Methods: PEG-citrate dendrimer and TiF4-dendrimer inclusion complex were synthesized and their molecular structures were evaluated using Fourier-transform Infrared Spectroscopy (FTIR), Hydrogen Nuclear Magnetic Resonance (HNMR), and Liquid Chromatography-Mass Spectrometry (LC-MS) tests. Forty-eight enamel samples were prepared and randomly divided into four groups: distilled water (negative control), TiF4 solution (T), dendrimer solution (D), and TiF4-dendrimer solution (TD). The microhardness of the samples was measured initially. Next, the samples underwent pH cycling, were exposed to the solutions, the microhardness was measured again, and microhardness loss was calculated. EDX analysis was performed on the surface and cross-sectional segments of the samples., Results: The microhardness loss was significantly higher in control (-65.1 ± 6.0) compared to other groups. No significant difference was observed between T (-47.9 ± 5.6) and D (-41.7 ± 12.0) and also D and TD (-40.5 ± 9.4) in this regard. Microhardness loss was significantly higher in T compared to TD group. The TD samples showed similar fluoride and titanium content in both surface and subsurface regions, while the T group had higher concentrations in the surface region. Moreover, the TD solution had a higher pH of 3.4 compared to the T solution's pH of 1.1., Conclusion: No significant difference was observed between the efficacy of TiF4-dendrimer and TiF4 solution in inhibiting demineralization while TiF4-dendrimer solution had the added advantage of having a higher pH., (© 2024 S. Karger AG, Basel.)

Purpose: To evaluate the effects of desensitizing MI Varnish (GC America) applied before or after bleaching on the mineral component of enamel and surface topography., Materials and Methods: The coronal portions of 10 freshly extracted bovine teeth were segmented for a total of 40 specimens. Enamel specimens obtained from each tooth were randomly divided into four groups (n = 10): Group A = no bleaching; Group B = bleaching with 40% hydrogen peroxide (HP); Group C = MI Varnish applied before bleaching; and Group D = MI Varnish applied after bleaching. The calcium (Ca) and phosphorus (P) levels of the specimens in each group were determined by energy dispersive spectroscopy (EDS). Morphologic changes were observed using scanning electron microscopy (SEM). One-way ANOVA and Tukey HSD tests were used for statistical analyses (α = .05)., Results: The mean Ca content of Group B was significantly lower than those of Groups A, C, and D (P < .05). The mean Ca content of Group C was significantly lower than that of Group A (P < .05). There was no significant difference in Ca content between the other groups (P > .05). The mean P content of Group A was significantly higher than those of Groups B to D (P < .05). There was no significant difference in P content between Groups B to D (P > .05)., Conclusions: Application of MI Varnish before or after in-office bleaching was effective in reducing mineral loss. However, applying MI Varnish after bleaching was more effective.

Objectives: The current research on single-nucleotide polymorphism (SNP) mutation sites at different positions of the FAM83H gene and their phenotypic changes leading to amelogenesis imperfecta (AI) is inconsistent. We identified a previously reported heterozygous nonsense mutation c.1192C>T (p.Q398*) in the FAM83H gene and conducted a comprehensive analysis of the dental ultrastructure and chemical composition changes induced by this mutation. Additionally, we predicted the protein feature affected by this mutation site. The aim was to further deepen our understanding of the diversity of AI caused by different mutation sites in the FAM83H gene., Methods: Whole-exome sequencing (WES) and Sanger sequencing were used to confirm the mutation sites. Physical features of the patient's teeth were investigated using various methods including cone beam computer tomography (CBCT), scanning electron microscopy (SEM), contact profilometry (roughness measurement), and a nanomechanical tester (nanoindentation measurement). The protein features of wild-type and mutant FAM83H were predicted using bioinformatics methods., Results: One previously discovered FAM83H heterozygous nonsense mutation c.1192C>T (p.Q398*) was detected in the patient. SEM revealed inconsistent dentinal tubules, and EDS showed that calcium and phosphorus were lower in the patient's dentin but higher in the enamel compared to the control tooth. Roughness measurements showed that AI patients' teeth had rougher occlusal surfaces than those of the control tooth. Nanoindentation measurements showed that the enamel and dentin hardness values of the AI patients' teeth were both significantly reduced compared to those of the control tooth. Compared to the wild-type FAM83H protein, the mutant FAM83H protein shows alterations in stability, hydrophobicity, secondary structure, and tertiary structure. These changes could underlie functional differences and AI phenotype variations caused by this mutation site., Conclusions: This study expands the understanding of the effects of FAM83H mutations on tooth structure., Clinical Relevance: Our study enhances our understanding of the genetic basis of AI and may contribute to improved diagnostics and personalized treatment strategies for patients with FAM83H-related AI., (© 2023. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

This study aimed to evaluate and compare the remineralisation, mechanical, anti-aging, acid resistance and antibacterial properties of calcium phosphate ion clusters (CPICs) materials with those of Duraphat and Icon. The remineralisation and mechanical properties were investigated using scanning electron microscopy, Fourier-transform infrared (FTIR) spectroscopy and nanoindentation. CPICs induced epitaxial crystal growth on the enamel surface, where the regrown enamel-like apatite layers had a similar hardness and elastic modulus to natural enamel (p > 0.05). Acid resistance and anti-aging properties were tested based on ion dissolution and surface roughness. CPICs exhibited similar calcium and phosphate ion dissolution to the control (p > 0.05), and its roughness decreased after thermocycling (p < 0.05), thereby decreasing the risk of enamel surface demineralisation. The minimum inhibitory concentration was 0.1 mg/ml, and the minimum bactericidal concentration ranged from 0.05 to 0.1 mg/ml. Overall, this biomimetic CPICs is a promising alternative to dental demineralisation., (© 2022 Australian Society of Endodontology Inc.)

Amelogenesis imperfecta (AI) is one of the typical dental genetic diseases in human. It can occur isolatedly or as part of a syndrome. Previous reports have mainly clarified the types and mechanisms of nonsyndromic AI. This review aimed to compare the phenotypic differences among the hereditary enamel defects with or without syndromes and their underlying pathogenic genes. We searched the articles in PubMed with different strategies or keywords including but not limited to amelogenesis imperfecta, enamel defects, hypoplastic/hypomaturation/hypocalcified, syndrome, or specific syndrome name. The articles with detailed clinical information about the enamel and other phenotypes and clear genetic background were used for the analysis. We totally summarized and compared enamel phenotypes of 18 nonsyndromic AI with 17 causative genes and 19 syndromic AI with 26 causative genes. According to the clinical features, radiographic or ultrastructural changes in enamel, the enamel defects were basically divided into hypoplastic and hypomineralized (hypomaturated and hypocalcified) and presented a higher heterogeneity which were closely related to the involved pathogenic genes, types of mutation, hereditary pattern, X chromosome inactivation, incomplete penetrance, and other mechanisms.The gene-specific enamel phenotypes could be an important indicator for diagnosing nonsyndromic and syndromic AI., (© 2023 Wiley Periodicals LLC.)

Proteomics has become an attractive method to study human and animal material, biological profile, and origin as an alternative to DNA analysis. It is limited by DNA amplification in ancient samples and its contamination, high cost, and limited preservation of nuclear DNA. Currently, three approaches are available to estimate sex-osteology, genomics, or proteomics, but little is known about the relative reliability of these methods in applied settings. Proteomics provides a new, seemingly simple, and relatively non-expensive way of sex estimation without the risk of contamination. Proteins can be preserved in hard teeth tissue (enamel) for tens of thousands of years. It uses two sexually distinct forms of the protein amelogenin in tooth enamel detectable by liquid chromatography-mass spectrometry; the protein amelogenin Y isoform is present in enamel dental tissue only in males, while amelogenin isoform X can be found in both sexes. From the point of view of archaeological, anthropological, and forensic research and applications, the reduced destruction of the methods used is essential, as well as the minimum requirements for sample size., (© 2023 Wiley-VCH GmbH.)

Objective: Dentin, enamel and the transition zone, called the dentin-enamel junction (DEJ), have an organization and properties that play a critical role in tooth resilience and in stopping the propagation of cracks. Understanding their chemical and micro-biomechanical properties is then of foremost importance. The aim of this study is to apply Brillouin microscopy on a complex biological structure, that is, the DEJ, and to compare these results with those obtained with Raman microscopy., Design: Both techniques allow noncontact measurements at the microscopic scale. Brillouin microscopy is based on the interaction between acoustic phonons and laser photons and gives a relation between the frequency shift of the scattered light and the stiffness of the sample. Raman spectra contain peaks related to specific chemical bonds., Results: Comparison of the Brillouin and Raman cartographies reveals correlations between mechanical and chemical properties. Indeed, the shapes of the phosphate content and stiffness curves are similar. The two spectroscopies give compatible values for the mean distance between two tubules, i.e., 4-6 µm. Moreover, for the first time, the daily cross striations of enamel could be studied, indicating a relationship between the variation in the phosphate concentration and the variation in the rigidity within the enamel prisms., Conclusions: We demonstrate here the possibility of using Brillouin scattering microscopy to both study complex biological materials such as the enamel-dentin junction and visualize secondary structures. Correlations between the chemical composition and mechanical properties could help in better understanding the tissue histology., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier Ltd. All rights reserved.)

Nitrogen isotopes are widely used to study the trophic position of animals in modern food webs; however, their application in the fossil record is severely limited by degradation of organic material during fossilization. In this study, we show that the nitrogen isotope composition of organic matter preserved in mammalian tooth enamel (δ 15 N enamel ) records diet and trophic position. The δ 15 N enamel of modern African mammals shows a 3.7‰ increase between herbivores and carnivores as expected from trophic enrichment, and there is a strong positive correlation between δ 15 N enamel and δ 15 N bone-collagen values from the same individuals. Additionally, δ 15 N enamel values of Late Pleistocene fossil teeth preserve diet and trophic level information, despite complete diagenetic loss of collagen in the same specimens. We demonstrate that δ 15 N enamel represents a powerful geochemical proxy for diet that is applicable to fossils and can help delineate major dietary transitions in ancient vertebrate lineages., (© 2023. The Author(s).)

Purpose: To evaluate bonding quality for orthodontic bracket bonding with different component combinations of self-etch primers in vitro., Methods: Metallic brackets were bonded to bovine lower incisors and assigned to groups. Group 1: comparison of self-etch (Transbond™ Plus, 3M™ Unitek, Neuss, Germany, n = 30; BrackFix® primer SE, VOCO®, Cuxhaven, Germany, n = 20) and etch-and-rinse bonding systems (Transbond™ XT, n = 20; BrackFix®, n = 20); group 2: comparison of different self-etch primer (Transbond™ Plus; BrackFix® primer SE) and adhesive (Transbond™ XT, n = 20; BrackFix®, n = 20) product combinations; group 3: testing cyclic fatigue bond strength of self-etch bonding systems (Transbond™ Plus, n = 20; BrackFix® primer SE, n = 20). All teeth were tested for shear bond strength according to the DIN-13990 standard, the adhesive remnant index (ARI) and enamel fractures were determined microscopically (10 × magnification)., Results: The mean shear bond strength of the self-etch (Transbond™ Plus: 16.38 ± 3.68 MPa; BrackFix® primer SE: 16.24 ± 1.73 MPa) and etch-and-rinse bonding systems (Transbond™ XT: 18.45 ± 2.56 MPa; BrackFix®: 17 ± 5.2 MPa) were of a clinically adequate order of magnitude (≥ 6-10 MPa) and were not statistically different. The component combination BrackFix® primer SE/Transbond™ XT adhesive led to a significantly lower shear bond strength (11.99 ± 3.68 MPa). There were no significant differences between static and fatigue shear bond strengths of self-etch bonding systems. Mean ARI scores mostly ranged between 4 and 5. The combination of the self-etch primer Transbond™ Plus with the BrackFix® adhesive led to a significantly increased enamel fracture rate., Conclusions: Based on the present findings bond strength of self-etch primers was equal to etch-and-rinse primers for bracket bonding. Combining different self-etch bonding systems might alter the clinical performance., (© 2021. Springer Medizin Verlag GmbH, ein Teil von Springer Nature.)

The purpose of this study was to examine the relationship between the bond strength of stainless steel with two types of resin cements (MMA- and composite-based) on bovine enamel depending on the directionality of the applied force. The specimens were either placed in water or subjected to thermal cycles (TC), and the shear or tensile bond strengths (SBS or TBS) were determined. The SBS showed significantly greater than the TBS for both types of cement, and the SBS and TBS for composite-based cement had larger than MMA-based one. No significant difference in SBS was observed in the cements even after being subjected to TC. Cohesive failures of the cement and bovine enamel in the composite-based group, while adhesive failures were observed in MMA-based one. Consequently, the direction of the force at both cements affected the retention of stainless steel, and MMA-based cement was preferred when prioritizing less enamel damages.

This study evaluated the etching pattern, surface microhardness, and bond strength for enamel and dentin submitted to treatment with phosphoric, glycolic, and ferulic acids. Enamel and dentin blocks were treated with phosphoric, glycolic, and ferulic acid to evaluate the surface and adhesive interface by scanning electron microscopy (2000×). Surface microhardness (Knoop) was evaluated before and after etching, and microtensile bond strength was evaluated after application of a two-step adhesive system (Adper Single Bond 2, 3M ESPE) at 24 hours and 12 months storage time points. Analysis of variance (ANOVA) and Tukey's test showed a decrease in the microhardness values for both substrates after application of each acid (p<0.0001). The reduction percentage was significantly higher for enamel treated with phosphoric acid (59.9%) and glycolic acid (65.1%) than for ferulic acid (16.5%) (p<0.0001), and higher for dentin that received phosphoric acid (38.3%) versus glycolic acid (27.8%) and ferulic acid (21.9%) (p<0.0001). Phosphoric and glycolic acids led to homogeneous enamel demineralization, and promoted the opening of dentinal tubules, whereas ferulic acid led to enamel surface demineralization and partially removed the smear layer. The adhesive-enamel interface showed micromechanical embedding of the adhesive in the interprismatic spaces when phosphoric and glycolic acids were applied. Ferulic acid showed no tag formation. Microtensile bond strength at both time points, and for both substrates, was lower with ferulic acid (p=0.0003/E; p=0.0011/D; Kruskal Wallis and Dunn). The bond strength for enamel and dentin decreased when using phosphoric and glycolic acids at the 12-month time evaluation (p<0.05). Glycolic acid showed an etching pattern and microhardness similar to that of phosphoric acid. Ferulic acid was not effective in etching the enamel or dentin, and it did not provide satisfactory bond strength to dental substrates., (© Operative Dentistry, 2023.)