Tooth acellular extrinsic fibre cementum incremental lines in humans are formed by parallel branched Sharpey's fibres and not by its mineral phase.

In humans, the growth pattern of the acellular extrinsic fibre cementum (AEFC) has been useful to estimate the age-at-death. However, the structural organization behind such a pattern remains poorly understood. In this study tooth cementum from seven individuals from a Mexican modern skeletal series were analyzed with the aim of unveiling the AEFC collagenous and mineral structure using multimodal imaging approaches. The organization of collagen fibres was first determined using: light microscopy, transmission electron microscopy (TEM), electron tomography, and plasma FIB scanning electron microscopy (PFIB-SEM) tomography. The mineral properties were then investigated using: synchrotron small-angle X-ray scattering (SAXS) for T-parameter (correlation length between mineral particles); synchrotron X-ray diffraction (XRD) for L-parameter (mineral crystalline domain size estimation), alignment parameter (crystals preferred orientation) and lattice parameters a and c; as well as synchrotron X-ray fluorescence for spatial distribution of calcium, phosphorus and zinc. Results show that Sharpey's fibres branched out fibres that cover and uncover other collagen bundles forming aligned arched structures that are joined by these same fibres but in a parallel fashion. The parallel fibres are not set as a continuum on the same plane and when they are superimposed project the AEFC incremental lines due to the collagen birefringence. The orientation of the apatite crystallites is subject to the arrangement of the collagen fibres, and the obtained parameter values along with the elemental distribution maps, revealed this mineral tissue as relatively homogeneous. Therefore, no intrinsic characteristics of the mineral phase could be associated with the alternating AEFC incremental pattern.

Acellular Extrinsic Fiber Cementum Is Invariably Present in the Superficial Layer of Apical Cementum in Mouse Molar.

The cementum is a highly mineralized tissue that covers the tooth root. The regional differences among the types of cementum, especially in the extrinsic fibers that contribute to tooth support, remain controversial. Therefore, this study used second harmonic generation imaging in conjunction with automated collagen extraction and image analysis algorithms to facilitate the quantitative examination of the fiber characteristics and the changes occurring in these fibers over time. Acellular extrinsic fiber cementum (AEFC) was invariably observed in the superficial layer of the apical cementum in mouse molars, indicating that this region of the cementum plays a crucial role in supporting the tooth. The apical AEFC exhibited continuity and fiber characteristics comparable with the cervical AEFC, suggesting a common cellular origin for their formation. The cellular intrinsic fiber cementum present in the inner layer of the apical cementum showed consistent growth in the apical direction without layering. This study highlights the dynamic nature of the cementum in mouse molars and underscores the requirement for re-examining its structure and roles. The findings of the present study elucidate the morphophysiological features of cementum and have broader implications for the maintenance of periodontal tissue health.

Synthesis, characterization, and evaluation of nano-hydroxyapatite based experimental calcium silicate cement as a root repair material / Síntesis, caracterización y evaluación de cemento experimental de silicato de calcio a base de nanohidroxiapatita como material de reparación radicular

Introduction: This study aimed to prepare a new root repair material including Portland cement, bismuth oxide, and nano-hydroxyapatite and analyze its physicochemical properties and its effects on the proliferation and differentiation of human dental pulp stem cells (hDPSCs). Material and Methods: Bismuth oxide as a radiopaque component and nano-hydroxyapatite particles were added to white Portland cement at 20% and 5% weight ratio, respectively. Characterization of the prepared cement was done using conventional methods. To examine the bioactivity of this new material, atomic absorption spectroscopy (AAS) was used for the investigation of the rate of calcium ions dissolution in simulated body fluid media. The viability of hDPSCs was assessed by an MTT assay after 1, 3 and 7 days. The odontogenic potential of this substance was evaluated by measuring alkaline phosphatase activity and alizarin red S staining. Results: Based on the bioactivity results, the cement presented high bio-activity, corroborating sufficiently with the calcium release patterns. The cell viability was significantly increased in new root repair material containing hydroxyapatite nanoparticles after 3 and 7 days (p<0.05). Conclusion: Moreover, alkaline phosphatase activity increased over 7 days in all experimental groups. The new cement containing nano-hydroxyapatite particles could be a good root repair material.

Objetivo: Este estudio tuvo como objetivo preparar un nuevo material de reparación de raíces que incluye cemento Portland, óxido de bismuto y nano-hidroxiapatita y analizar sus propiedades fisicoquímicas y sus efectos sobre la proliferación y diferenciación de células madre de pulpa dental humana. Material y Métodos: El óxido de bismuto como compo-nente radiopaco y las partículas de nano-hidroxiapatita se agregaron al cemento Portland blanco en una proporción en peso del 20 % y el 5 %, respectivamente. La caracterización del cemento preparado se realizó utilizando métodos con-vencionales. Para examinar la bioactividad de este nuevo material, se utilizó la espectroscopia de absorción atómica para investigar la velocidad de disolución de los iones de calcio en medio fluido corporal simulado. La viabilidad de las células madre de pulpa dental humana se evaluó mediante un ensayo MTT después de 1, 3 y 7 días. El potencial odontogénico de esta sustancia se evaluó midiendo la actividad de la fosfatasa alcalina y la tinción con rojo de alizarina S.Resultados: Con base en los resultados de bioactividad, el cemento presentó alta bioactividad, corroborando suficientemente con los patrones de liberación de calcio. La viabilidad celular aumentó significativamente en el nuevo material de reparación de raíces que contenía nanopartículas de hidroxiapatita después de 3 y 7 días (p<0,05). Conclusión: Además, la actividad de la fosfatasa alcalina aumentó durante 7 días en todos los grupos experimentales. El nuevo cemento que contiene partículas de nanohidroxiapatita podría ser un buen material de reparación radicular.

Immunohistochemical Evaluation of Periodontal Regeneration Using a Porous Collagen Scaffold.

(1) Aim: To immunohistochemically evaluate the effect of a volume-stable collagen scaffold (VCMX) on periodontal regeneration. (2) Methods: In eight beagle dogs, acute two-wall intrabony defects were treated with open flap debridement either with VCMX (test) or without (control). After 12 weeks, eight defects out of four animals were processed for paraffin histology and immunohistochemistry. (3) Results: All defects (four test + four control) revealed periodontal regeneration with cementum and bone formation. VCMX remnants were integrated in bone, periodontal ligament (PDL), and cementum. No differences in immunohistochemical labeling patterns were observed between test and control sites. New bone and cementum were labeled for bone sialoprotein, while the regenerated PDL was labeled for periostin and collagen type 1. Cytokeratin-positive epithelial cell rests of Malassez were detected in 50% of the defects. The regenerated PDL demonstrated a larger blood vessel area at the test (14.48% ± 3.52%) than at control sites (8.04% ± 1.85%, p = 0.0007). The number of blood vessels was higher in the regenerated PDL (test + control) compared to the pristine one (p = 0.012). The cell proliferative index was not statistically significantly different in pristine and regenerated PDL. (4) Conclusions: The data suggest a positive effect of VCMX on angiogenesis and an equally high cell turnover in the regenerated and pristine PDL. This VCMX supported periodontal regeneration in intrabony defects.

Analysis of the nano and microstructures of the cervical cementum and saliva in periodontitis: A pilot study.

OBJECTIVES: During the progression of periodontitis, the structures of the cementum and saliva are altered due to pathological changes in the environment. This study aimed to analyze the nanostructures of the cervical cementum and saliva in patients with periodontitis. METHODS: Patients with periodontitis (n = 10) and periodontally healthy controls (n = 8) were included. Single-rooted teeth with indications for extraction were obtained from individuals. The cervical-thirds of the roots were sectioned transversely to obtain 1 mm thick sections. Unstimulated whole saliva samples were collected from each individual. The nanostructures of the cementum and saliva were analyzed using small and wide-angle X-ray scattering methods. RESULTS: The mean radius and distance values of the cementum nanoparticles in the periodontitis and control groups were 368 Å and 1152 Å, and 377 Å and 1186 Å, respectively. The mean radius and distance values of the saliva nanoparticles in the periodontitis and control groups were 425 Å and 1359 Å, and 468 Å and 1452 Å, respectively. More wide-angle X-ray scattering profile peaks were observed in the cementum of the controls. Similarities were observed between the 3D profiles of the cementum and the saliva nanoparticles. CONCLUSIONS: According to the results of the present study, (i) the cementum and saliva nanoparticles were of similar size in periodontitis and healthy controls, (ii) the cementum was more crystalline according to the (002) crystallographic plane in controls, and (iii) the similarities in the 3D-profile of the cementum and saliva nanoparticles suggest some interactions between them in the sulcus/periodontal pocket at the nanolevel.

Comparative evaluation of different human dental tissues and alveolar bone for DNA quantity and quality for forensic investigation.

In this study, the efficacy of dental tissues (cementum, dentine and pulp) and alveolar bone as a potential source of DNA was tested in terms of the quality and quantity using nuclear and mitochondrial markers for forensic investigation.This study found dentine as the best source of DNA with only 5.36% imbalanced (PHR<0.7) heterozygous loci. Pulp showed the highest quantity of DNA but exhibited 22.3% imbalanced (PHR<0.7) heterozygous loci. Cementum with highest (46.67%) heterozygote imbalance proved to be the last choice as a source of DNA. Alveolar bone exhibited the second-highest total yield of DNA/mg of tissue. All Global Filer™ STR loci were amplified in 70% samples of fresh alveolar bone whereas for 30% samples, only partial profile was generated along with successful sex determination. All the dental tissues and alveolar bone samples amplified non STR markers (D-loop, Cytochrome Oxidase I, SRY, AMEL). Of the alveolar bones from archival samples, one sample exhibited full STR profile whereas other alveolar bone samples gave partial profiles. This study substantiates alveolar bone as an alternate source of nuclear and mitochondrial DNA.

Effect of proximal box elevation on fracture resistance and microleakage of premolars restored with ceramic endocrowns.

BACKGROUND: Restoration of endodontically treated premolar is in high risk for biomechanical failure, and often presents with subgingival margins. Proximal box elevation (PBE) has been used to relocate subgingival cavity outlines. OBJECTIVE: To evaluate the influence of PBE on fracture resistance and gingival microleakage of premolars with endodontic access cavities following ceramic endocrown. METHODS: Eighty sound maxillary premolars with standardized Class II cavities on mesial surfaces were randomly assigned to four groups (n = 20 in each group). Groups E1, E2 and E3, with proximal margins located in dentin/cementum, 2 mm below the cemento-enamel junction (CEJ), simulated subgingival location. Group E4 (supragingival group), with proximal margins located in enamel, 1 mm above the CEJ, was used as the positive control. For margin elevation of the proximal cavities, bulk-fill Smart Dentin Replacement (SDR), a visible light cured resin composite, was applied in group E1, and conventional resin composite (3M Z350 XT, a light-activated composite) was placed in group E2. Group E3 was only treated with a ceramic crown and served as the negative control. In all groups, computer-aided design (CAD) ceramic endocrowns were adhesively inserted, and fracture resistance, failure mode and microleakage were evaluated. RESULTS: A higher fracture resistance value was observed in PBE groups E1 and E2, regardless of the materials used (P = 0.038, and 0.010, respectively, vs E3), and fracture resistance in group E1 was higher than that in group E2. In teeth without PBE, the percentage of catastrophic failures reached 70%. Compared to group E3, a lower frequency distribution of microleakage was detected in supragingival group E4 (P = 0.031). No increased percentage of microleakage was observed in groups treated with PBE. CONCLUSION: For endodontically treated maxillary premolars restored with ceramic endocrowns, PBE increases fracture resistance but not microleakage.

FOS Rearrangement and Expression in Cementoblastoma.

Cementoblastomas are rare odontogenic tumors developing in close proximity to the roots of teeth. Due to their striking morphologic resemblance to osteoblastomas of the peripheral skeleton, we set out to determine whether cementoblastomas harbor the same FOS rearrangements with overexpression of c-FOS as has recently been described for osteoblastomas. In total, 16 cementoblastomas were analyzed for FOS expression by immunohistochemistry and for FOS rearrangements by fluorescence in situ hybridization (FISH). We observed strong and diffuse staining of c-FOS in 71% of cementoblastomas and identified a FOS rearrangement in all cases (n=3) applicable for FISH. In the remaining cases, FISH failed due to decalcification. Cementoblastomas harbor similar FOS rearrangements and show overexpression of c-FOS like osteoblastomas, suggesting that both entities might represent parts of the spectrum of the same disease.

A minimally destructive protocol for DNA extraction from ancient teeth.

Ancient DNA sampling methods-although optimized for efficient DNA extraction-are destructive, relying on drilling or cutting and powdering (parts of) bones and teeth. As the field of ancient DNA has grown, so have concerns about the impact of destructive sampling of the skeletal remains from which ancient DNA is obtained. Due to a particularly high concentration of endogenous DNA, the cementum of tooth roots is often targeted for ancient DNA sampling, but destructive sampling methods of the cementum often result in the loss of at least one entire root. Here, we present a minimally destructive method for extracting ancient DNA from dental cementum present on the surface of tooth roots. This method does not require destructive drilling or grinding, and, following extraction, the tooth remains safe to handle and suitable for most morphological studies, as well as other biochemical studies, such as radiocarbon dating. We extracted and sequenced ancient DNA from 30 teeth (and nine corresponding petrous bones) using this minimally destructive extraction method in addition to a typical tooth sampling method. We find that the minimally destructive method can provide ancient DNA that is of comparable quality to extracts produced from teeth that have undergone destructive sampling processes. Further, we find that a rigorous cleaning of the tooth surface combining diluted bleach and UV light irradiation seems sufficient to minimize external contaminants usually removed through the physical removal of a superficial layer when sampling through regular powdering methods.

Parturitions, menopause and other physiological stressors are recorded in dental cementum microstructure.

The life history pattern of recent humans is uniquely derived in many of its aspects including an extended post-reproductive lifespan combined with short interbirth intervals. A number of theories have been proposed to explain the evolution of this unusual pattern. However most have been difficult to test due to the fragmentary nature of the hominin fossil record and the lack of methods capable of inferring such later life history events. In search of a method we tested the hypothesis that the physiologically impactful events of parturition and menopause are recorded in dental cementum microstructure. We performed histomorphological analyses of 47 teeth from 15 individuals with known life history events and were able to detect reproductive events and menopause in all females. Furthermore, we found that other stressful events such as systemic illnesses and incarceration are also detectable. Finally, through the development of a novel analytical method we were able to time all such events with high accuracy (R-squared = 0.92).

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.

Combined dental wear and cementum analyses in ungulates reveal the seasonality of Neanderthal occupations in Covalejos Cave (Northern Iberia).

We propose for the first time the use of the combination of two high-resolution techniques, dental wear (meso- and microwear) and dental cementum analyses, to gain a better understanding of Neanderthal subsistence strategies and occupational patterns. Dental wear analysis provides information not only on ungulate palaeodiet and palaeoenvironments but also on hunting time and seasons. Dental cementum analysis allows the accurate determination of the age and season at death of a prey. Our study has focused on the Cantabrian region and has applied both methods to investigate the Mousterian faunal assemblages in Covalejos Cave. Identification of the ungulate palaeodiet reveals information on the environmental conditions of the studied region. Moreover, it may facilitate observation on the evolution of both palaeodiet and palaeoenvironment throughout the site sequence. Results show a general stability in the palaeoenvironmental conditions and in the ungulate palaeodiet throughout the Mousterian sequence; this finding may be attributed to the role of the area as a climate refuge, and slight differences in levels 8, 7 and 4 suggest long- or short-term but repeated Neanderthal occupations at different seasons in the annual cycle.

Fracture resistance and the mode of failure produced in metal-free crowns cemented onto zirconia abutments in dental implants.

The purpose of the investigation was to analyze fracture resistance and mode of failure of zirconium oxide (zirconia) abutments placed on dental implants bearing crowns of different esthetic materials: zirconia, lithium disilicate (LDS), and nano-ceramic resin, for replacing single teeth in the anterior sector. Eighty implant-abutment-crown units were divided into four groups: Group T-MC (control): 20 metal-ceramic crowns cemented onto titanium abutments; Group Z-Z: 20 zirconia crowns on zirconia abutments; Group Z-LD: 20 lithium disilicate crowns on zirconia abutments; and Group Z-NCR: 20 nano-ceramic resin crowns on zirconia abutments. Specimens underwent a fatiguing process (dynamic loading and thermocycling), followed by static loading to evaluate mechanical fracture resistance, and the mode of failure produced. Mean fracture resistance values were: Control Group T-MC, 575.85±120.01 N; Group Z-Z 459.64±66.52 N; Group Z-LD, 531.77±34.10 N; and Group Z-NCR, 587.05±59.27 N. In Group T-MC, fracture occurred in the prosthetic fixing screw in 100% of specimens. In Group Z-Z, 80% of fractures occurred in the fixing screw, 15% in the abutment, and 5% in the abutment and crown. In Group Z-LD, 60% of fractures were produced in the fixing screw and 40% in the abutment. In Group Z-NCR, 70% of fractures were produced in the fixing screw and 30% in the abutment. All the abutments and crowns analyzed have the potential to withstand the physiological occlusal forces to which they would be subject in the anterior region. Lithium disilicate and nano-ceramic resin crowns cemented onto zirconia abutments are a good restoration alternative for single implants in the anterior sector.

Age at death estimation by cementochronology: Too precise to be true or too precise to be accurate?

OBJECTIVES: Cementochronology based on annual deposition of acellular cementum is acknowledged for its superior performance for estimating age-at-death but is also disregarded because of its suspicious effectiveness. This article aims to provide a standardized framework for the validation of the technique and to define and test cementochronology's performance and limitations. MATERIALS AND METHODS: To determine the boundaries of the cementum aging technique, we applied a certified protocol on a sample of 200 healthy canines from individuals of known age, sex, postmortem, and inhumation intervals from anthropological and anatomical collections. We scored readability and preservation of cementum and measured the agreement between estimates, i.e., the precision, and assessed the quality of the accordance between estimates and chronological age, i.e., the accuracy. To investigate the applicability on ancient material, 200 additional canines extracted from archeological assemblages were included. Accuracy and precision were analyzed for each age group in considering postmortem intervals and taphonomical conditions. RESULTS: A strong correlation was found between chronological age and estimates (r = .927; p = .000) but results reveal an association between readability of incremental lines and chronological age (p < .05) and a notable difference in both precision and accuracy between individuals under and over 50 years. Results also demonstrate that taphonomy can be a serious obstacle increasing imprecision by a factor of three. DISCUSSION: Improperly adopted, cementochronology can lead to precise but inaccurate estimations. If methodological, physiological, and taphonomical factors are taken into account, then, and only then, cementochronology will serve as a versatile and powerful tool for age-at-death estimation.

An unusual disordered alveolar bone material in the upper furcation region of minipig mandibles: A 3D hierarchical structural study.

Teeth are subjected to compressive loads during mastication. Under small loads the soft tissue periodontal ligament (PDL) deforms most. However when the loads increase and the PDL is highly compressed, the tooth and the alveolar bone supporting the tooth, begin to deform. Here we report on the structure of this alveolar bone in the upper furcation region of the first molars of mature minipigs. Using light microscopy and scanning electron microscopy (SEM) of bone cross-sections, we show that this bone is hypermineralized, containing abundant small pores around 1-5 µm in diameter, lacunae around 10-20 µm as well as larger spaces. This bone does not possess the typical lamellar motif or other repeating structures normally found in cortical or trabecular mammalian bone. We also use high resolution focused ion beam scanning electron microscopy (FIB-SEM) in the serial surface mode to image the 3D organization of the demineralized bone matrix. We show that the upper furcation bone matrix has a disordered isotropic structure composed mainly of individual collagen fibrils with no preferred orientation, as well as highly staining material that is probably proteoglycans. Much larger aligned arrays of collagen fibers - presumably Sharpey's fibers - are embedded in this material. This unusual furcation bone material is similar to the disordered material found in human lamellar bone. In the upper furcation region this disordered bone comprises almost all the volume excluding Sharpey's fibers. We surmise that this most unusual bone type functions to resist the repeating compressive loads incurred by molars during mastication.

Influence of pre-heating and ceramic thickness on physical properties of luting agents.

PURPOSE: The objective of this in vitro study was to evaluate the influence of ceramic thickness and pre-heating of luting agents on their physical properties. MATERIALS AND METHODS: The materials RelyX Arc, RelyX Ultimate, RelyX Veneer, and Filtek Z350 Flow were handled at different temperatures (23°C or 54°C), inserted into matrix, and photoactivated through ceramic disks (0.75 mm or 1.5 mm). The following tests were performed ( n=8): degree of conversion, Knoop Hardness, cross-link density, water sorption, solubility, and ultimate tensile strength. Data were analyzed using three-way analysis of variance and Tukey's test (α=0.05). RESULTS: Regarding ceramic thickness, the thinnest ceramic resulted in higher values of Knoop Hardness ( p=0.027). The lowest temperature (23°C) resulted in a higher solubility ( p=0.0257), and water sorption ( p=0.0229) values. There was also statistical difference among the materials: RelyX Arc showed a higher degree of conversion and ultimate tensile strength, followed by RelyX Veneer, RelyX Ultimate, and Filtek Z350 Flow. For Knoop Hardness and cross-link density tests, RelyX Ultimate showed the highest values, followed by RelyX Arc, RelyX Veneer, and Filtek Z350 Flow. For water sorption and solubility, RelyX Veneer showed the highest values, followed by RelyX Arc, RelyX Ultimate, and Filtek Z350 Flow. CONCLUSION: Pre-heating interfered with water sorption and solubility, whereas ceramic thickness only affected Knoop Hardness; the physical properties of the materials are dependent on their composition.

The mineral content of the hard dental tissue of mesiodens.

OBJECTIVE: Mesiodens is the most common form of supernumerary tooth mainly located between the maxillary central incisors. Its etiology is not completely understood but both genetic and environmental factors are assumed. The degree of mineralization and inorganic element content in hard tooth tissues is poorly understood as well as is the durability and suitability for allo- and auto-transplantation. Therefore aim of this study was to examine the content of inorganic elements. MATERIALS AND METHODS: This study included 26 mesiodens teeth and 26 normal central incisor teeth as controls. All specimens were prepared for SEM/EDS analysis which was aimed at specific sites on the enamel, dentine and cementum in order to evaluate the weight percentage and ratio of important inorganic elements. RESULTS: and Conclusion. The results showed that there was a difference in the weight percentage of selected inorganic elements (calcium, phosphorus, oxygen, carbon, magnesium and sodium) in all three types of dental hard tissues but the differences were mostly expressed in the cementum tissue. The statistical analysis showed that the differences were marginally significant especially for calcium and phosphorus values and ratio in the enamel and dentine. The carbon and magnesium content in all three hard tissues showed the most differences, but overall, the hard tissues mineral content of the mesiodens did not differs significantly from healthy teeth.

Isolation of DNA from small amounts of elephant ivory: Sampling the cementum with total demineralization extraction.

Genotyping ivory samples can determine the geographic origin of poached ivory as well as the legality of ivory being sold in ivory markets. We conducted a series of experiments to determine where the DNA is most concentrated in ivory samples and how best to increase DNA yield from groups of samples likely to vary in DNA concentration. We examined variation in DNA amplification success from: the layer(s) of the tusk (cementum and/or dentine) being extracted, demineralization temperature and time, and the concentration of eluates. Since demineralization of the pulverized sample produces a pellet and supernatant, we also assessed DNA amplification success from the pellet, the supernatant, their combination, as well as variation in the respective amounts used for extraction. Our results show that the outer cementum layer of the tusk contains the highest concentration of DNA and should be separated and used exclusively as the source material of ivory processed for extraction, when available. Utilizing the combined demineralized lysate improves extraction efficiency, as does increasing demineralization time to 3 or more days, conducted at 4°C. The most significant improvements occurred for low template DNA ivory samples followed by medium quality samples. Amplification success of high quality samples was not affected by these changes. Application of this optimized method to 3068 ivory samples resulted in 81.2% of samples being confirmed for both alleles at a minimum of 10 out of 16 microsatellite loci, which is our threshold for inclusion in DNA assignment analyses.

Incremental distribution of strontium and zinc in great ape and fossil hominin cementum using synchrotron X-ray fluorescence mapping.

Cementum and the incremental markings it contains have been widely studied as a means of ageing animals and retrieving information about diet and nutrition. The distribution of trace elements in great ape and fossil hominin cementum has not been studied previously. Synchrotron X-ray fluorescence (SXRF) enables rapid scanning of large tissue areas with high resolution of elemental distributions. First, we used SXRF to map calcium, phosphorus, strontium and zinc distributions in great ape dentine and cementum. At higher resolution, we compared zinc and strontium distributions in cellular and acellular cementum in regions where clear incremental markings were expressed. We then mapped trace element distributions in fossil hominin dentine and cementum from the 1.55-1.65 million year old site of Koobi Fora, Kenya. Zinc, in particular, is a precise marker of cementum increments in great apes, and is retained in fossil hominin cementum, but does not correspond well with the more diffuse fluctuations observed in strontium distribution. Cementum is unusual among mineralized tissues in retaining so much zinc. This is known to reduce the acid solubility of hydroxyapatite and so may confer resistance to resorption by osteoclasts in the dynamic remodelling environment of the periodontal ligament and alveolar bone.

Cementum as a source of DNA in challenging forensic cases.

Each forensic case is characterized by its own uniqueness. Deficient forensic cases require additional sources of human identifiers to assure the identity. We report on two different cases illustrating the role of teeth in answering challenging forensic questions. The first case involves identification of an adipocere male found in a car submersed in water for approximately 2 years. The second scenario, which involves paternity DNA testing of an exhumed body, was performed approximately 2.8 years post-mortem. The difficulty in anticipating the degradation of the DNA is one of the main obstacles. DNA profiling of dental tissues, DNA quantification by using real-time PCR (PowerQuant™ System/Promega) and a histological dental examination have been performed to address the encountered impediments of adverse post-mortem changes. Our results demonstrate that despite the adverse environmental conditions, a successful STR profile of DNA isolated from the root of teeth can be generated with respect to tooth type and apportion. We conclude that cementocytes are a fruitful source of DNA. Cementum resists DNA degradation in comparison to other tissues with respect to the intra- and inter-individual variation of histological and anatomical structures.