Extracting the truth through chemical analyses: Early life histories of Victorian-era dental patients in Aotearoa New Zealand.

OBJECTIVES: There are few bioarcheological analyses of life experiences in colonial period Aotearoa New Zealand, despite this being a time of major adaptation and social change. In our study, early life histories are constructed from multi-isotope and enamel peptide analysis of permanent first molars associated with Victorian era dental practices operating between AD 1881 and 1905 in Invercargill. Chemical analyses of the teeth provide insight into the childhood feeding practices, diet, and mobility of the people who had their teeth extracted. MATERIALS AND METHODS: Four permanent left mandibular first molars were analyzed from a cache of teeth discovered at the Leviathan Gift Depot site during excavations in 2019. The methods used were: (1) enamel peptide analysis to assess chromosomal sex; (2) bulk (δ13 Ccarbonate ) and incremental (δ13 Ccollagen and δ15 N) isotope analysis of dentin to assess childhood diet; and (3) strontium (87 Sr/86 Sr) and oxygen (δ18 O) isotope analysis of enamel to assess childhood residency. Two modern permanent first molars from known individuals were analyzed as controls. RESULTS: The archaeological teeth were from three chromosomal males and one female. The protein and whole diets were predominately based on C3 -plants and domestic animal products (meat and milk). A breastfeeding signal was only identified in one historic male. All individuals likely had childhood residences in Aotearoa. DISCUSSION: Unlike most bioarcheological studies that rely on the remains of the dead, the teeth analysed in this study were extracted from living people. We suggest that the dental patients were likely second or third generation colonists to Aotearoa, with fairly similar childhood diets. They were potentially lower-class individuals either living in, or passing through, the growing colonial center of Invercargill.

Micro-punches versus micro-slices for serial sampling of human dentine: Striking a balance between improved temporal resolution and measuring additional isotope systems.

RATIONALE: The last decade has seen a dramatic increase in the application of serial sampling of human dentine in archaeology. Rapid development in the field has provided many improvements in the methodology, in terms of both time resolution as well as the ability to integrate more isotope systems in the analysis. This study provides a comparison of two common sampling approaches, allowing researchers to select the most suitable approach for addressing specific research questions. METHODS: Two common approaches for sequential sampling of human dentine (micro-punches and micro-slices) are compared in terms of viability and efficacy. Using archaeological deciduous second molars and permanent first molars, this study demonstrates how the two approaches capture aspects of the weaning process in different ways. In addition, different aspects related to the extraction protocols, such as the thickness of the central slide and the solubilisation step, are also evaluated. RESULTS: While both approaches show similar intra-tooth isotopic patterns, the micro-punches approach is preferable for research that requires a very fine temporal resolution, while the micro-slices approach is best for research where δ34 S values are needed, or when the samples are poorly preserved. In addition, the solubilisation step has a large effect on collagen yield, and, to a lesser extent, on isotopic compositions. Therefore, it is important to ensure that only samples that have undergone the same pre-treatment protocol are directly compared. CONCLUSIONS: We present the pros and cons of the two micro-sampling approaches and offer possible mitigation strategies to address some of the most important issues related to each approach.

Studies of Peculiar Mg-Containing and Oscillating Bioapatites in Sheep and Horse Teeth.

New types of biological apatites have been discovered in molar sheep and horse teeth and are divided in two types. In the first and more general type, the release of Mg ions is parallel to the changes in composition of apatite leading to a final stoichiometric ratio of Ca to P ions, going from dentin depth towards the boundary of enamel with air. Inside dentin, another apatite sub-types were discovered with alternating layers of Mg-rich and C-rich apatites. The approximate formal stoichiometric relationships for these peculiar types of bioapatites are suggested. We identified two kinds of ion-exchanges responsible for formation of peculiar apatites. Various combinations of main and minor elements lead to new versions of biological apatites.

Novel Approach to Tooth Chemistry. Quantification of the Dental-Enamel Junction.

The dentin-enamel junction (DEJ) is known for its special role in teeth. Several techniques were applied for the investigation of the DEJ in human sound molar teeth. The electron (EPMA) and proton (PIXE) microprobes gave consistent indications about the variability of elemental concentrations on this boundary. The locally increased and oscillating concentrations of Mg and Na were observed in the junction, in the layer adhering to the enamel and covering roughly half of the DEJ width. The chemical results were compared with the optical profiles of the junction. Our chemical and optical results were next compared with the micromechanical results (hardness, elastic modulus, friction coefficient) available in the world literature. A strong correlation of both result sets was proven, which testifies to the self-affinity of the junction structures for different locations and even for different kinds of teeth and techniques applied for studies. Energetic changes in tooth strictly connected with crystallographic transformations were calculated, and the minimum energetic status was discovered for DEJ zone. Modeling of both walls of the DEJ from optical data was demonstrated. Comparing the DEJ in human teeth with the same structure found in dinosaur, shark, and alligator teeth evidences the universality of dentin enamel junction in animal world. The paper makes a contribution to better understanding the joining of the different hard tissues.

[Determination of Allyl Isothiocyanate in Mustard and Horseradish Extracts by Single Reference GC and HPLC Based on Relative Molar Sensitivities].

The main component of the Mustard and Horseradish extracts, which are used as natural food additives in Japan, is allyl isothiocyanate (AITC). The determination of AITC using GC-FID is the official method employed in the quality control assessments for these products. In this method, a commercially available AITC reagent is used as a calibrant. However, 1H-quantitative NMR (qNMR) analysis revealed that the AITC reagents contain impurity. Therefore, we examined the GC-FID and HPLC-refractive index detector (LC-RID) method based on relative molar sensitivities (RMSs) to high-purity single reference (SR). The RMSs of AITC/SR under the GC-FID and LC-RID conditions were accurately determined using qNMR. The AITC in two types of food additives was quantified using qNMR, SR GC-FID, and SR LC-RID methods. Both SR GC-FID and SR LC-RID showed good agreement within 2% with the AITC content determined by direct qNMR.

Sensing platform for pico-molar level detection of ethyl parathion using Au-Ag nanoclusters based enzymatic strategy.

This work demonstrates a simple, cost effective and ultrasensitive detection of ethyl parathion, an organophosphorus (OPs) pesticide, using enzyme based fluorometric sensing strategy by employing bimetallic BSA@AuAg nanoclusters (NC). The sensing assay is based on the "quenched off" state of bimetallic NC with the addition of Cu2+ ions that can be "switched on" due to generation of thiocholine (TCh), a catalytic product of enzymatic reaction of acetylthiocholine (ATCh) using acetylcholinesterase (AChE) enzyme. The generated TCh preferably seize Cu2+ ions from BSA@AuAg NC-Cu2+ ensemble and recovered the fluorescence of BSA@AuAg NC. The presence of ethyl parathion can be monitored optically due to its inhibitory action towards AChE enzyme leading to suppression of thiocholine (TCh) formation and subsequently decreases TCh-Cu2+ interaction that ultimately retrieved quenched off state of bimetallic NC. The synthesized biosensor is appropriate for the ultrasensitive sensing of ethyl parathion in pM range, exhibiting 2.40 pM as lowest limit of detection (LOD) which is the least known so far. Further, the real sample analysis adds on for the appropriateness of the synthesized nanoprobe by depicting excellent reproducibility and robustness. The designed assay proved its specificity towards pesticides in general and ethyl parathion in particular when employed with other commonly used non-OPs pesticides.

Physiochemical characteristics: A robust tool to overcome teeth heterogeneity on predicting laser ablation profile.

To avoid excessive tissue removal and collateral damage, the high-power density laser is apt for dental surgery also need to have high precision. For high-precision dental surgery with minimal tissue damage, the present work frames a method to predict laser ablation profile based on surface morphology and chemical composition of dentin. The surface morphology and chemical composition were studied on different dentin samples using scanning electron microscope (SEM) and Energy Dispersive X-ray Analysis (EDAX), respectively. The key laser ablation parameters (ω0 , Deff , and Fth ) were determined by laser irradiation study using 800 nm, Ti:Sapphire femtosecond laser at processing condition of 100 fs, 10 kHz and 10 mm/s. The dentin samples show a strong linear correlation between physiochemical characteristics and laser ablation parameters. The surface morphology exhibits a negative linear correlation with threshold fluence, whereas the converse is true for chemical composition. The laser ablation parameters of a random dentin sample are derived from the knowledge of linearity data. From the obtained laser ablation parameters, the complete theoretical ablation profile is constructed and validated with experimental ablation profile. Even though the surface morphology of dentin shows high linearity, the concentration of Ca and P can be used as the most feasible probe in clinical settings. Furthermore, the laser ablation rate and ablation efficiency are predicted by the method to optimize the laser processing condition for any specific teeth. The versatility of the method overcomes the problem of heterogeneity on various teeth and simplifies the method of finding optimal laser processing condition for immaculate laser surgery.

Early specialized maritime and maize economies on the north coast of Peru.

We assess diet and economies of middle Holocene (∼7,500 to 4,000 calibrated [cal] B.P.) humans at coexisting mound sites (Huaca Prieta and Paredones) in north coastal Peru and document regular consumption of maize by ∼6,500 to 6,000 cal B.P. and its earliest use as a staple food in this area of the Andes between 5,000 and 4,500 cal B.P. Stable isotope data from enamel carbonates and dentin collagen (childhood diet) and dental microwear texture analysis (adult diet) demonstrate dietary and economic specialization. Previous studies revealed maize and mixed-food refuse at both sites, but this study documents actual food consumption, showing that these communities situated a few hundred meters apart had significantly distinct diets in childhood and adulthood. Huaca Prieta focused on marine resources, although there are some contributions from terrestrial meat. Paredones individuals primarily consumed maize during childhood (up to 70% of the juvenile diet), as shown by δ13C values, apatite-collagen spacing, and discriminant analysis of δ13Ccoll, δ13Ccarb, and δ15N values. Maize was likely used as a weaning food (e.g., gruel and/or chicha-a maize beverage), hinting at the significant role of breastfeeding mothers, weanling infants, and children in the development of maize as a staple crop. Additionally, dental microwear data show Paredones adult diets are high in abrasives, potentially from maize processing. The distinct foodways at these neighboring sites result from and also reflect their social and political distinctions. These differences in food production, distribution, and consumption generated opportunities for exchange, an interaction that bound them together in mutual benefit.

Osteogenic Potential of Decellularized Tooth Matrix.

We studied the possibility of using decellularized tooth matrix as a scaffold to restore bone tissue. It was found that mesenchymal stem cells underwent spontaneous osteogenic differentiation on the decellularized tooth matrix, which makes it possible to use it as a natural allograft in the treatment of resorption of alveolar bone tissue.

Near-Surface Studies of the Changes to the Structure and Mechanical Properties of Human Enamel under the Action of Fluoride Varnish Containing CPP-ACP Compound.

Changes to the features of the enamel surface submitted to induced demineralisation and subsequent remineralisation were studied. The in vitro examination was conducted on polished slices of human molar teeth, divided in four groups: the untreated control (n = 20), challenged by a demineralisation with orthophosphoric acid (H3PO4) (n = 20), and challenged by a demineralisation following remineralisation with fluoride (F) varnish containing casein phosphopeptides (CPP) and amorphous calcium phosphate (ACP) compounds (n = 20). The specimens' enamel surfaces were subjected to analysis of structure, molecular arrangement, mechanical features, chemical composition, and crystalline organization of apatite crystals. Specimens treated with acid showed a significant decrease in crystallinity, calcium, and phosphorus levels as well as mechanical parameters, with an increase in enamel surface roughness and degree of carbonates when compared to the control group. Treatment with fluoride CPP-ACP varnish provided great improvements in enamel arrangement, as the destroyed hydroxyapatite structure was largely rebuilt and the resulting enamel surface was characterised by greater regularity, higher molecular and structural organisation, and a smoother surface compared to the demineralised one. In conclusion, this in vitro study showed that fluoride CPP-ACP varnish, by improving enamel hardness and initiating the deposition of a new crystal layer, can be an effective remineralising agent for the treatment of damaged enamel.

The petrous bone: Ideal substrate in legal medicine?

Over the last few years, palaeogenomic studies of the petrous bone (the densest part of the temporal bone) have shown that it is a source of DNA in both larger quantities and of better quality than other bones. This dense bone around the otic capsule has therefore been called the choice substrate in palaeogenomics. Because the practice of forensic genetics responds to different imperatives, we implemented a study aimed at (i) understanding how and why the petrous bone is an advantageous substrate in ancient DNA studies and (ii) establishing whether it is advantageous in forensic STR typing. We selected 50 individual skeletal remains and extracted DNA from one tooth and one petrous bone from each. We then amplified 24 STR markers commonly used in forensic identification and compared the quality of that amplification using the RFU intensities of the signal as read on the STR profiles. We also performed histological analyses to compare (i) the microscopic structure of a petrous bone and of a tooth and (ii) the microscopic structure of fresh petrous bone and of an archaeological or forensic sample. We show that the RFU intensities read on STR profiles are systematically higher in experiments using DNA extracted from petrous bones rather than teeth. For this reason, we were more likely to obtain a complete STR profile from petrous bone material, increasing the chance of identification in a forensic setting. Histological analyses revealed peculiar microstructural characteristics (tissue organization), unique to the petrous bone, that might explain the good preservation of DNA in that substrate. Therefore, it appears that despite the necessity of analysing longer fragments in forensic STR typing compared to NGS palaeogenomics, the use of petrous bones in forensic genetics could prove valuable, especially in cases involving infants, toothless individuals or very degraded skeletal remains.

The dental proteome of Homo antecessor.

The phylogenetic relationships between hominins of the Early Pleistocene epoch in Eurasia, such as Homo antecessor, and hominins that appear later in the fossil record during the Middle Pleistocene epoch, such as Homo sapiens, are highly debated1-5. For the oldest remains, the molecular study of these relationships is hindered by the degradation of ancient DNA. However, recent research has demonstrated that the analysis of ancient proteins can address this challenge6-8. Here we present the dental enamel proteomes of H. antecessor from Atapuerca (Spain)9,10 and Homo erectus from Dmanisi (Georgia)1, two key fossil assemblages that have a central role in models of Pleistocene hominin morphology, dispersal and divergence. We provide evidence that H. antecessor is a close sister lineage to subsequent Middle and Late Pleistocene hominins, including modern humans, Neanderthals and Denisovans. This placement implies that the modern-like face of H. antecessor-that is, similar to that of modern humans-may have a considerably deep ancestry in the genus Homo, and that the cranial morphology of Neanderthals represents a derived form. By recovering AMELY-specific peptide sequences, we also conclude that the H. antecessor molar fragment from Atapuerca that we analysed belonged to a male individual. Finally, these H. antecessor and H. erectus fossils preserve evidence of enamel proteome phosphorylation and proteolytic digestion that occurred in vivo during tooth formation. Our results provide important insights into the evolutionary relationships between H. antecessor and other hominin groups, and pave the way for future studies using enamel proteomes to investigate hominin biology across the existence of the genus Homo.

The effect of hydrochloric acid (HCl) on permanent molars: A scanning electron microscope (SEM) and energy dispersive X-ray spectroscopy (EDS) study.

It is well established that acid disposal is a potentially effective method used by criminal syndicates to hinder the identification of victims. This study documents the effects of continuous immersion in hydrochloric acid (HCl, 37%) on molars using macroscopic analysis, scanning electron microscopy (SEM), and energy dispersive X-ray spectroscopy (EDS). The goal of this study is to aid in distinguishing visually unrecognizable fragments of dental remains when drastic changes in morphology have occurred as a result of acid exposure. Macroscopic, SEM, and EDS analysis were conducted on seven maxillary molars before and after HCl treatment. Molars reduced in weight relative to the length of time immersed in HCl and the dissolution time was over 40 hours longer than reported in previous studies, at just over 66 hours. SEM and EDS analysis showed acid-treated teeth exhibited morphological patterns such as cracking and layering visible at high magnification. Calcium/phosphorous ratios fell within the expected range of 1.6-2.5, indicating that HCl-treated teeth are still identifiable as osseous or dental tissue even when not visually identifiable as teeth. This is the first study to present SEM images of molar cementum before and after immersion in HCl and to present EDS results. This information can assist researchers and investigators in determining the presence of dental tissue in a forensic context associated with acid disposal.

Expression, localization and synthesis of small leucine-rich proteoglycans in developing mouse molar tooth germ.

The gene expression and protein synthesis of small leucine-rich proteoglycans (SLRPs), including decorin, biglycan, fibromodulin, and lumican, was analyzed in the context of the hypothesis that they are closely related to tooth formation. In situ hybridization, immunohistochemistry, and organ culture with metabolic labeling of [35S] were carried out in mouse first molar tooth germs of different developmental stages using ICR mice at embryonic day (E) 13.5 to postnatal day (P) 7.0. At the bud and cap stage, decorin mRNA was expressed only in the surrounding mesenchyme, but not within the tooth germ. Biglycan mRNA was then expressed in the condensing mesenchyme and the dental papilla of the tooth germ. At the apposition stage (late bell stage), both decorin and biglycan mRNA were expressed in odontoblasts, resulting in a switch of the pattern of expression within the different stages of odontoblast differentiation. Decorin mRNA was expressed earlier in newly differentiating odontoblasts than biglycan. With odontoblast maturation and dentin formation, decorin mRNA expression was diminished and localized to the newly differentiating odontoblasts at the cervical region. Simultaneously, biglycan mRNA took over and extended its expression throughout the new and mature odontoblasts. Both mRNAs were expressed in the dental pulp underlying the respective odontoblasts. At P7.0, both mRNAs were weakly expressed but maintained their spatial expression patterns. Immunostaining showed that biglycan was localized in the dental papillae and pulp. In addition, all four SLRPs showed clear immunostaining in predentin, although the expressions of fibromodulin and lumican mRNAs were not identified in the tooth germs examined. The organ culture data obtained supported the histological findings that biglycan is more predominant than decorin at the apposition stage. These results were used to identify biglycan as the principal molecule among the SLRPs investigated. Our findings indicate that decorin and biglycan show spatial and temporal differential expressions and play their own tissue-specific roles in tooth development.

Mercury and selenium concentrations in fishes of the Upper Colorado River Basin, southwestern United States: A retrospective assessment.

Mercury (Hg) and selenium (Se) are contaminants of concern for fish in the Upper Colorado River Basin (UCRB). We explored Hg and Se in fish tissues (2,324 individuals) collected over 50 years (1962-2011) from the UCRB. Samples include native and non-native fish collected from lotic waterbodies spanning 7 major tributaries to the Colorado River. There was little variation of total mercury (THg) in fish assemblages basin-wide and only 13% (272/1959) of individual fish samples exceeded the fish health benchmark (0.27 µg THg/g ww). Most THg exceedances were observed in the White-Yampa tributary whereas the San Juan had the lowest mean THg concentration. Risks associated with THg are species specific with exceedances dominated by Colorado Pikeminnow (mean = 0.38 and standard error ± 0.08 µg THg/g ww) and Roundtail Chub (0.24 ± 0.06 µg THg/g ww). For Se, 48% (827/1720) of all individuals exceeded the fish health benchmark (5.1 µg Se/g dw). The Gunnison river had the most individual exceedances of the Se benchmark (74%) whereas the Dirty Devil had the fewest. We identified that species of management concern accumulate THg and Se to levels above risk thresholds and that fishes of the White-Yampa (THg) and Gunnison (Se) rivers are at the greatest risk in the UCRB.

Application of Trace Elemental Profile of Known Teeth for Sex and Age Estimation of Ajnala Skeletal Remains: a Forensic Anthropological Cross-Validation Study.

Teeth store crucial information needed for medical, environmental, genomics, public health, and forensic anthropological research work. The prolonged forensic utility of teeth is ensured by their extended postmortem longevity as they can resist almost all sorts of chemical, biological, and physical degradations or destructions. The main aim of the present investigation was to utilize the discriminant functions and regression equations generated from the elemental profile of known teeth for estimating sex and age of unknown human skeletal remains excavated from an abandoned well at Ajnala (Amritsar, India). The written records mentioned that 282 Indian mutineer soldiers were killed in July 1857, their cadavers were dumped in the disused well, and then a religious structure was built over the periphery of the said well. These human remains, along with some contextual items, were excavated non-scientifically in April 2014. Preliminary results obtained from application of different forensic anthropological techniques like stable isotope, pulp-tooth area ratio, and mtDNA analyses have indicated that these remains belonged to adult males. In present study, the elemental concentrations of 100 mandibular molars of known age and sex were estimated from wavelength-dispersive X-ray fluorescence spectrophotometer (WD-XRF) analyzer. The statistical equations so generated from elemental concentrations of known teeth were applied to estimate the probable age and the sex of unknown mandibular teeth (N = 100) collected from Ajnala skeletal assemblage. The elements Pb and As were detected in ancient teeth only whereas the detection of elements like Ba, Se, and Te was limited to modern teeth samples only. When the statistical equations so generated were applied to elemental concentrations of Ajnala teeth, it was found that 96% teeth belonged to adult males and the remaining ones were classified to be that of females. Though sexual differences were observed in concentrations of majority of elements, statistically significant differences were found in elemental concentrations of very few teeth. Age estimates of unknown teeth were found in the age ranges of 19 to 48 years. Thus, the trace elemental analysis results supported the written records that the victims were adult males. The cross-validated application of elemental profiles of known teeth for establishing the identity of unknown teeth is the first forensic anthropological study reported from India. Though the obtained accuracy levels were not found within acceptable forensic threshold limits, the present study results may guide future researches involving human hard tissues. It may be concluded that trace elemental concentrations of teeth may be influenced by the factors like age and sex of an individual and thus cannot be used for accurate and reliable forensic sex or age estimations. Dental trace elemental composition can be used as a forensic tool only if used in conjunction with other morphological or molecular analysis of the unknown dental remains.

Atomic level observation and structural analysis of phosphoric-acid ester interaction at dentin.

The functional monomer 10-methacryloyloxydecyl dihydrogen phosphate (10-MDP), used in many dental adhesives, self-assembles in nano-layers at adhesive-tooth interfaces. Recently, several states of the POH groups of 10-MDP_Ca salts were suggested, while their actual status has not been elucidated yet. We mechanistically investigated 10-MDP_Ca-salt nano-layering at adhesive-dentin interfaces, correlatively using scanning transmission electron microscopy with energy-dispersive X-ray spectrometry (STEM-EDS), X-ray diffraction (XRD) and solid state nuclear magnetic resonance (NMR). STEM-EDS confirmed the presence of Ca and P in each nano-layer. Both XRD and NMR revealed that the two terminal POH groups of 10-MDP reacted with Ca. This twofold POH interaction of 10-MDP with Ca was stable in water and is therefore expected to contribute to durable nano-layering of 10-MDP_Ca salts in the hybrid and adhesive layer. STATEMENT OF SIGNIFICANCE: The functional monomer 10-methacryloyloxydecyl dihydrogen phosphate (10-MDP), commonly used in dental adhesives with favorable long-term clinical outcome, has been documented to self-assemble into nano-layers at adhesive-tooth interfaces. Characterizing ultra-morphologically (STEM) and chemically (STEM-EDS, XRD, NMR) the mechanisms of interaction of 10-MDP with bulk dentin in a similar manner as what occurs clinically, it was found that the water stable 10-MDP_Ca salts consist of CaRPO4, meaning that the two OH groups of the phosphate group of 10-MDP ionically reacted with Ca. This stable structure is expected to contribute to durable nano-layering of 10-MDP_Ca salts in the hybrid and adhesive layer and hence to clinical longevity of the adhesively bonded tooth restoration.

A Novel Etchant System for Orthodontic Bracket Bonding.

Orthodontic treatment is widely used to correct irregular teeth and/or jaw discrepancies to improve oral function and facial aesthetics. However, it is frequently associated with enamel damage that include chipping, demineralisation, and white spot formation. So far, current bonding systems that can maintain shear bond strengths (SBS) suitable for clinical performance are unable to limit enamel demineralisation, adhesive remnants and damage caused on removal of brackets after treatment. This study reports a novel "safe enamel etch" clinically viable procedure that was accomplished via application of novel etchant pastes developed with ß-tricalcium phosphate and monocalcium phosphate monohydrate powders mixed with citric acid (5 M) or phosphoric acid (37% PA) to yield BCA and BPA etchants respectively. Although enamel etched with clinically used PA gel yielded higher SBS than the BCA/BPA etchants, it exhibited greater adhesive remnants with evidence of enamel damage. In contrast, the experimental etchants resulted in unblemished enamel surfaces with zero or minimal adhesive residue and clinically acceptable SBS. Furthermore, the BPA etchant caused lower enamel decalcification with extensive calcium-phosphate precipitation. The study conclusively showed that BPA facilitated in vitro enamel adhesion without detrimental effects of the aggressive PA gel with potential for remineralisation and saving time at the post-debonding step.

Nanoscale properties and deformation of human enamel and dentin.

This research uses quasi-static nanoindentation and nanoscratching to quantify human tooth deformation as a function of enamel rod and dentin tubule orientations at the nanoscale. Nanoindentation tests were performed on enamel and dentin to determine elastic modulus, hardness, and observe fracture. Additionally, nanoscratch tests were performed to determine pileup geometry and parameters such as recovery, scratch hardness, and scratch roughness. In enamel, it was found that nanofiber orientation gives rise to unique microcrack propagation and nanofiber behavior that affect these properties. For dentin, densification and organic content affect these properties.

Using ImageJ (Fiji) to Analyze and Present X-Ray CT Images of Enamel.

X-ray micro CT has become a popular methodology for the nondestructive analysis of dental tissues and has been used extensively in the amelogenesis field. The aim of this chapter is to introduce ImageJ/Fiji to researchers new to CT scanning and the analysis of CT image data. The program can be applied to analyzing X-ray CT images of enamel but can be extrapolated to other tissues as well.