Analysis of Covarine Particle in Toothpaste Through Microfluidic Simulation, Experimental Validation, and Electrical Impedance Spectroscopy.

Covarine, copper phthalocyanine, a novel tooth whitening ingredient, has been incorporated into various toothpaste formulations using diverse technologies such as larger flakes, two-phase pastes, and microbeads. In this study, we investigated the behavior of covarine microbeads (200 µm) in Colgate advanced white toothpaste when mixed with artificial and real saliva. Our analysis utilized a custom-designed microfluidic mixer with 400 µm wide channels arranged in serpentine patterns, featuring a Y-shaped design for saliva and toothpaste flow. The mixer, fabricated using stereolithography 3D printing technology, incorporated a flexible transparent resin (Formlabs' Flexible 80A resin) and PMMA layers. COMSOL simulations were performed by utilizing parameters extracted from toothpaste and saliva datasheets, supplemented by laboratory measurements, to enhance simulation accuracy. Experimental assessments encompassing the behavior of covarine particles were conducted using an optical profilometer. Viscosity tests and electrical impedance spectroscopy employing recently developed all-carbon electrodes were employed to analyze different toothpaste dilutions. The integration of experimental data from microfluidic chips with computational simulations offers thorough insights into the interactions of covarine particles with saliva and the formation of microfilms on enamel surfaces.

Influence of casein phosphopeptide-amorphous calcium phosphate on the surface topography and composition of nickel-titanium archwires during orthodontic treatment with fixed appliances.

PURPOSE: To investigate the surface topography and nickel content of nickel-titanium (NiTi) archwires exposed to either routine oral hygiene or a prophylactic regimen with casein phosphopeptide-amorphous calcium phosphate (CPP-ACP) during orthodontic treatment. METHODS: This in vivo study involved 40 orthodontic patients with fixed appliances, who were randomly assigned to either a routine oral hygiene group or a CPP-ACP supplementary regimen group. Twenty new NiTi archwires served as controls. All archwires underwent scanning electron microscopy and energy-dispersive spectroscopy to evaluate their surface topography and elemental composition. The nickel content was quantified as a percentage of total weight and the Ni/Ti ratio, and statistical comparisons were made using pairwise tests. RESULTS: Wires exposed to fluoride toothpaste showed signs of pitting corrosion, deep grooves, and corrosion debris. In contrast, wires exposed to supplementary CPP-ACP exhibited smooth surface areas interspersed with microdefects and deposits. Statistically significant differences in nickel content were found between the new and retrieved archwires, as well as between wires exposed to routine oral hygiene and CPP-ACP (P < 0.001). The archwires exposed to CPP-ACP had the lowest nickel content (P < 0.001). CONCLUSION: The use of CPP-ACP holds promise for application as a safe anticariogenic agent with possible protective properties during orthodontic treatment.

Oral micro-electronic platform for temperature and humidity monitoring.

Intraoral theranostics, the integration of diagnostics and therapeutics within the oral cavity, is gaining significant traction. This pioneering approach primarily addresses issues like xerostomia (dry mouth), commonly resulting from cancer treatment, with a specific focus on monitoring temperature and humidity. This paper introduces the innovative Intra-Oral Portable Micro-Electronic (IOPM) fluidic theranostic device platform. It leverages conventional dental spoons by incorporating advanced sensors for precise measurements of oral temperature and humidity. Personalization options include a microfluidic chip and a tooth model, enabling targeted delivery of therapeutic agents to optimize treatment outcomes. The electronic control system simplifies the administration of fluid dosages, intelligently adjusted based on real-time oral cavity temperature and humidity readings. Rigorous experimental evaluations validate the platform's precision in delivering fluid volumes at predefined intervals. This platform represents a transformative advancement for individuals contending with oral health challenges such as xerostomia (dry mouth). Furthermore, it has the potential to elevate oral healthcare standards by providing advanced diagnostics and tailored therapeutic solutions, benefiting both patients and dental professionals alike.

Toothpaste ingestion-evaluating the problem and ensuring safety: systematic review and meta-analysis.

This systematic review and meta-analysis aimed to evaluate the ingestion of toothpaste and its sequelae. The study adhered to the PRISMA guidelines and was registered in the PROSPERO database. A comprehensive search strategy was conducted across multiple databases, resulting in the inclusion of 18 relevant publications. Eligible studies encompassed various designs and included both children and adults as the study population. Data extraction was carried out systematically, and relevant information on study characteristics, interventions, and outcomes were collected. The assessment of bias was performed using the Joanna Briggs Institute's Critical Appraisal Tools showing variations of bias among the included studies. The overall risk of systemic toxicity was found to be low, and no severe or life-threatening events were reported in the included studies. Furthermore, some toothpaste formulations containing higher concentrations of fluoride were associated with an increased risk of dental fluorosis. These findings have several implications for practice and policy. Healthcare providers and dental professionals should emphasize the importance of promoting safe toothpaste use, especially in vulnerable populations such as young children who are more prone to accidental ingestion. Public health campaigns and educational initiatives should aim to raise awareness about appropriate toothpaste usage and the potential risks. In addition, toothpaste manufacturers and regulatory bodies should consider revising guidelines and regulations to ensure the safety of oral care products, including the appropriate concentration of active ingredients. Future research should focus on investigating the long-term effects of toothpaste ingestion, exploring potential interactions between different active ingredients, and evaluating the efficacy of current preventive measures.

The role of cadmium in the pathogenesis of myeloid leukemia in individuals with anemia, deficiencies in vitamin D, zinc, and low calcium dietary intake.

Iron deficiency, vitamin D deficiency and low calcium diet are frequent health problems with severe long- term consequences. Upon absorption from the duodenum, cadmium binds to transferrin, and cells with the highest density of transferrin receptor 1 (TfR1) take up the majority of the circulating cadmium. Nowadays, it is clear that individuals with iron deficiency anemia have increased blood levels of cadmium because of higher absorption rate, mediated by divalent metal transporter 1 (DMT1). However, the transient receptor potential vanilloid receptor 6 (TRPV6), known as a calcium carrier, is able to bind and transport cadmium as well. In the case of low calcium diet or vitamin D deficiency, TRPV6 may be overexpressed in the intestine and kidney tubules and absorbs (re-uptake in the case of renal tubules) cadmium in larger quantities, resulting in an increased cadmium blood levels. We speculate that the final event in the case of low calcium dietary diet and/or vitamin D deficiency is similar to what is observed in the case of iron deficiency, that cells with the highest levels of TfR1 (for example, megakaryocyte/erythrocyte progenitors and pro-erythroblasts) take up most of the circulating cadmium, which is powerful malignancy inductor, leading to appearance of acute myeloid leukemia (AML).

The Participation of Pediatric Dentists in Oral Health Promotion and Education in Serbia.

Objectives: To identify the involvement of Serbian paediatric dentists in oral health promotion and education, and to propose further actions for the improvement of these activities. Methods: This is an analysis of data collected by a cross-sectional, questionnaire-based survey of 445 dentists involved in the provision of dental health services to children at the primary healthcare level. We explored dentists' involvement in oral health education and promotion and the cooperation with other health professionals at the healthcare centre and the community level as well as their attitudes towards the importance of some factors influencing their work. Results: Dentists estimate their cooperation with different services with ratings higher than 3 on the scale of 1 to 5. They reported the highest satisfaction in cooperation with paediatric services for preschool and schoolchildren (4.0±1.0). At the community level, they reported excellent cooperation with kindergartens (4.4±0.8), while collaboration with Roma health mediators (3.14±1.34) and nongovernmental organizations (2.5±1.4) received lower ratings. According to the average rating (4.7±0.7), dentists perceive the motivation of patients and/or their guardians for keeping good oral health as the factor with the highest importance for the quality of interventions they provide. Conclusions: Dentists involved in the provision of dental healthcare for children and adolescents in primary healthcare centres in Serbia participate in different oral healthcare education and promotion activities in the community, and highlight the importance of strengthening cooperation with healthcare and other professionals and services aimed at vulnerable population groups, both within the health sector and nongovernmental organizations.

3D-Printed Microfluidic Chip for Real-Time Glucose Monitoring in Liquid Analytes.

The connection of macrosystems with microsystems for in-line measurements is important in different biotechnological processes as it enables precise and accurate monitoring of process parameters at a small scale, which can provide valuable insights into the process, and ultimately lead to improved process control and optimization. Additionally, it allows continuous monitoring without the need for manual sampling and analysis, leading to more efficient and cost-effective production. In this paper, a 3D printed microfluidic (MF) chip for glucose (Glc) sensing in a liquid analyte is proposed. The chip made in Poly(methyl methacrylate) (PMMA) contains integrated serpentine-based micromixers realized via stereolithography with a slot for USB-like integration of commercial DropSens electrodes. After adjusting the sample's pH in the first micromixer, small volumes of the sample and enzyme are mixed in the second micromixer and lead to a sensing chamber where the Glc concentration is measured via chronoamperometry. The sensing potential was examined for Glc concentrations in acetate buffer in the range of 0.1-100 mg/mL and afterward tested for Glc sensing in a cell culturing medium. The proposed chip showed great potential for connection with macrosystems, such as bioreactors, for direct in-line monitoring of a quality parameter in a liquid sample.

A novel microfluidic compact disc to investigate electrochemical property changes between artificial and real salivary samples mixed with mouthwashes using electrical impedance analysis.

Diagnosing oral diseases at an early stage may lead to better preventive treatments, thus reducing treatment burden and costs. This paper introduces a systematic design of a microfluidic compact disc (CD) consisting of six unique chambers that run simultaneously from sample loading, holding, mixing and analysis. In this study, the electrochemical property changes between real saliva and artificial saliva mixed with three different types of mouthwashes (i.e. chlorhexidine-, fluoride- and essential oil (Listerine)-based mouthwashes) were investigated using electrical impedance analysis. Given the diversity and complexity of patient's salivary samples, we investigated the electrochemical impedance property of healthy real saliva mixed with different types of mouthwashes to understand the different electrochemical property which could be a foundation for diagnosis and monitoring of oral diseases. On the other hand, electrochemical impedance property of artificial saliva, a commonly used moisturizing agent and lubricant for the treatment of xerostomia or dry mouth syndrome was also studied. The findings indicate that artificial saliva and fluoride-based mouthwash showed higher conductance values compared to real saliva and two other different types of mouthwashes. The ability of our new microfluidic CD platform to perform multiplex processes and detection of electrochemical property of different types of saliva and mouthwashes is a fundamental concept for future research on salivary theranostics using point-of-care microfluidic CD platform.

Enamel incremental markings in the deciduous teeth of children from the Early Bronze and modern ages.

OBJECTIVE: The purpose of this study was to determine whether children from Early Bronze Age and modern populations differ in terms of the width of the neonatal line (NNL) and the occurrence of accentuated lines in enamel. DESIGN: The sample (N = 59) consisted of two groups: 29 deciduous teeth removed from the jaws of children (dental age range from 1 to 10 years) whose skeletal remains were found in Early Bronze archaeological graves in Mokrin Serbia, and 30 present-day exfoliated deciduous teeth from 6 to 11 year old children. Mothers, whose children participated in this study, provided information regarding their health during pregnancy. The analysis was carried out on ground sections with a scanning electron microscope. Two clinicians measured the width of the NNL and counted the accentuated lines in the enamel. RESULTS: There was a statistically significant difference between the children from the two groups regarding the width of the NNL. The width of the NNL between children whose mothers were healthy and diagnosed with gestational diabetes was significantly different. Most subjects did not have accentuated lines in the prenatal enamel, regardless of whether they were from the Bronze or Modern age. Accentuated lines were dominantly found in the postnatal enamel of the children from the Early Bronze age. CONCLUSIONS: This study is the first to investigate the width of the NNL in teeth of Maros children and Serbian children from the modern age. The wider NNL of children from the Early Bronze age indicates the possibility that they have experienced more overall stress in perinatal life.

Role of Nanomaterials in the Fabrication of bioNEMS/MEMS for Biomedical Applications and towards Pioneering Food Waste Utilisation.

bioNEMS/MEMS has emerged as an innovative technology for the miniaturisation of biomedical devices with high precision and rapid processing since its first R&D breakthrough in the 1980s. To date, several organic including food waste derived nanomaterials and inorganic nanomaterials (e.g., carbon nanotubes, graphene, silica, gold, and magnetic nanoparticles) have steered the development of high-throughput and sensitive bioNEMS/MEMS-based biosensors, actuator systems, drug delivery systems and implantable/wearable sensors with desirable biomedical properties. Turning food waste into valuable nanomaterials is potential groundbreaking research in this growing field of bioMEMS/NEMS. This review aspires to communicate recent progress in organic and inorganic nanomaterials based bioNEMS/MEMS for biomedical applications, comprehensively discussing nanomaterials criteria and their prospects as ideal tools for biomedical devices. We discuss clinical applications for diagnostic, monitoring, and therapeutic applications as well as the technological potential for cell manipulation (i.e., sorting, separation, and patterning technology). In addition, current in vitro and in vivo assessments of promising nanomaterials-based biomedical devices will be discussed in this review. Finally, this review also looked at the most recent state-of-the-art knowledge on Internet of Things (IoT) applications such as nanosensors, nanoantennas, nanoprocessors, and nanobattery.

Sex and age determination of human mandible using anthropological parameters and TCI and Kvaal methods: study of a Serbian medieval sample.

PURPOSE: The aim of the study was to test the efficacy, reliability, and applicability of the Kvaal and Tooth Coronal Index (TCI) age determination techniques, and then to compare them with each other, as well as with the conventional anthropological age and sex determination techniques. METHODS: The analyzed material originates from the medieval necropolis of the Vinca-Belo brdo site. During the research, 60 periapical (PA) and 30 orthopantomographic (OPT) images were analyzed. On each analyzed tooth, age assessment was performed using both TCI and Kvaal techniques. The obtained values of dental estimated age were compared with age estimated by anthropological analysis, and the deviations between the estimated and chronological age were analyzed in relation to the assessment technique, type of dental radiograph, tooth group, sex, and age. RESULTS: The mean error between TCI and the osteological method was 8.44 (SD = 7.56, Min = 0.169, Max = 36.4) and between Kvaal and the osteological method was 7.71 (SD = 5.57, Min = 0.133, Max = 26.7). The average value of age recorded by TCI method was 32.5 years and by Kvaal method was 34.7 years. There was no statistically significant difference based on the two radiographic methods, gender, individual teeth, or tooth group pairs. There was a statistically significant positive correlation between age and the error present. CONCLUSION: Gender determination based only on the mandible has a high correlation with the anthropological gender determination. The Kvaal method and the TCI method have proven their efficiency, reliability, and applicability. Significant correlation has been observed between dental and anthropological age and sex determination methods.

Oral Health in 12- and 15-Year-Old Children in Serbia: A National Pathfinder Study.

The aim of the paper is to present the oral health profile of 12- and 15-year-old schoolchildren in Serbia. Basic Methods for Oral Health Surveys of the WHO were implemented to record: Decayed, Missing, and Filled Teeth/Surfaces Index (DMFT/DMFS), gingival bleeding, enamel fluorosis and other structural anomalies, dental erosion, dental trauma, and oral mucosal lesions. In addition, Silness and Löe plaque index and orthodontic status were assessed. A total of 36% of 12-year-olds and 22% of 15-year-olds in Serbia were caries-free. The mean DMFT was 2.32 ± 2.69 for 12-year-olds and 4.09 ± 3.81 for 15-year-olds. DMFT was made up largely by the decayed component. Gingival bleeding was present in 26% of examined 12-year-old and 18% of 15-year-old children. Dental plaque was observed in 63% of both 12- and 15-year-olds. Fluorosis, structural anomalies, dental erosion, dental trauma, and oral mucosal lesion were rarely detected. Low prevalence of malocclusions was found. Oral disease is still a common public health problem among schoolchildren in Serbia. A significant increase in the prevalence of caries disease between 12- and 15-year-old groups implies that preventive care for adolescents requires special attention. Corrective actions and reforms to the current school-based oral health prevention program are needed to further improve oral health in Serbian children.

Determination of salivary concentrations of leptin and adiponectin, ability to reduce ferric ions and total antioxidant capacity of saliva in patients with severe early childhood caries.

Introduction: One of the most common oral diseases affecting children is early childhood caries (ECC). The link between oxidative stress and ECC has been proven in numerous clinical studies. Technical and biological variability were so high in most of the studies that none of the markers have yet been proven suitable for routine clinical use. This study aimed to evaluate the antioxidant status and the levels of leptin and adiponectin in saliva of children with severe early childhood caries (S-ECC). Methods: Morning unstimulated saliva samples were collected from children (n = 40, 0-6 years old) for the evaluation of oxidative stress which were measured by total antioxidant capacity (TAC), and by the ferric reducing antioxidant power (FRAP) assays, as well as to assess the salivary levels of leptin and adiponectin. FRAP, TAC, leptin and adiponectin concentrations were evaluated in S-ECC group (n = 31) and caries free group CF (n = 9). All results were analyzed based on age and sex. Results: Overall median salivary leptin and adiponectin levels were 5.59 pg/mL and 24.86 ng/mL, respectively. Significantly lower leptin levels were observed in saliva of caries free children (4.66 pg/mL) than in the S-ECC group (6.64 pg/mL, p < 0.01). No significant difference was observed for adiponectin levels (S-ECC and CF, 25.31 and 23.2 ng/mL, respectively, p = 0.961). TAC and FRAP values of saliva had similar values in children with S-ECC and caries free children. TAC and FRAP values also remained stable with the age of the children, without significant differences with respect to sex. Conclusion: The increased concentrations of leptin in saliva of children with S-ECC suggests that leptin may play a role in inflammatory and immune responses in the development of early childhood caries.

The Measurement of Contact Angle, pH, and Conductivity of Artificial Saliva and Mouthwashes on Enamel, Glass-Ionomer, and Composite Dental Materials.

The aim of this study was to tackle the topic of appropriate recommendations for artificial-saliva and mouthwash usage. The contact angle, pH, and conductivity of two artificial saliva solutions, four mouthwashes, and their mixtures on enamel, glass-ionomer, and composite dental materials were measured. The measurements were conducted with a MATLAB algorithm to minimize human error. The obtained values for the contact angle were in the range from 7.98° to 52.6°, and they showed completely nonlinear and nonuniform behavior for all investigated liquids and on all investigated substrates. Results reveal statistically significant differences among all tested liquids on all investigated substrates (p < 0.05). pH values ranged from 1.54 to 7.01. A wide range of conductivity values were observed, from 1205µS/cm in the saliva-stimulating solution to 6679 mS/cm in the artificial saliva. Spearman's test showed a moderate positive correlation between the pH and conductivity of the tested fluids (R = 0.7108). A comparison of the data obtained using Image J software and the MATLAB algorithm showed consistency, not exceeding 5% error. When an experiment uses human material and bioactive materials THAT are used in biomedicine as substrates, an additional definition of protocols is highly recommended for future research on this topic.

Performance Evaluation of Dental Flosses Pre- and Post-Utilization.

Dental floss is an oral hygiene product used to remove food and plaque in places where toothbrushes cannot reach. Even though over the years since its introduction some research in suitable materials has been performed, thread cracking and wear can still compromise efficiency. The aim of this study was to examine the morphological properties of four different commercially available dental floss types before and after use. For that purpose, scanning electron microscopy and optical microscopy were used to assess the flosses before and after use, and tension testing was performed to determine any degradation in the floss performance after utilization. The analyzed floss samples verify the hypothesis that the properties of the floss need to be known in depth, before recommending a specific type to patients for daily use in all clinical indications.

Vascular Complications in Individuals with Type 2 Diabetes Mellitus Additionally Increase the Risk of Femoral Neck Fractures Due to Deteriorated Trabecular Microarchitecture.

Individuals with diabetes mellitus type 2 (T2DM) have an increased risk of hip fracture, especially if vascular complications are present. However, microstructural origins of increased bone fragility in T2DM are still controversial. DXA measurement of the contralateral hip and three-dimensional microCT analyses of femoral neck trabecular microarchitecture were performed in 32 individuals (26 women and 6 men, 78 ± 7 years). The specimens were divided to two groups: T2DM individuals with hip fracture (DMFx, n = 18) and healthy controls (CTL, n = 14). DMFx group consisted of individuals with vascular complications (DMFx_VD, n = 8) and those without vascular complications (DMFx_NVD, n = 10). T-score was significantly lower in DMFx_VD and DMFx_NVD than in controls (p < 0.001). BV/TV, Tb.N, Tb.Sp, SMI, and FD varied among DMFx_NVD, DMFx_VD, and CTL groups (p = 0.023, p = 0.004, p = 0.008, p = 0.001, p = 0.007, respectively). Specifically, BV/TV of DMFx_VD was significantly lower than that of DMFx_NVD group (p = 0.020); DMFx_NVD group had higher Tb.N and lower Tb.Sp compared with DMFx_VD (p = 0.006, p = 0.012, respectively) and CTL (p = 0.026, p = 0.035, respectively). DMFx group and healthy controls showed similar BV/TV, Tb.Th, Tb.N, Tb.Sp, Conn.D, DA, and FD (p = 0.771, p = 0.503, p = 0.285, p = 0.266, p = 0.208, p = 0.235, p = 0.688, respectively), while SMI was significantly higher in controls (p = 0.005). Two distinct phenotypes of bone fragility were identified in T2DM patients: patients with vascular complications showed impaired trabecular microarchitecture, whereas bone fragility in the group without vascular complications was independent on trabecular microarchitecture pattern. Such heterogeneity among T2DM patients may explain contradicting literature data and may set a basis for further studies to evaluate fracture risk related to T2DM.

New perspectives on cytoskeletal dysregulation and mitochondrial mislocalization in amyotrophic lateral sclerosis.

Amyotrophic lateral sclerosis (ALS) is a progressive neurodegenerative disease characterized by selective, early degeneration of motor neurons in the brain and spinal cord. Motor neurons have long axonal projections, which rely on the integrity of neuronal cytoskeleton and mitochondria to regulate energy requirements for maintaining axonal stability, anterograde and retrograde transport, and signaling between neurons. The formation of protein aggregates which contain cytoskeletal proteins, and mitochondrial dysfunction both have devastating effects on the function of neurons and are shared pathological features across several neurodegenerative conditions, including ALS, Alzheimer's disease, Parkinson's disease, Huntington's disease and Charcot-Marie-Tooth disease. Furthermore, it is becoming increasingly clear that cytoskeletal integrity and mitochondrial function are intricately linked. Therefore, dysregulations of the cytoskeletal network and mitochondrial homeostasis and localization, may be common pathways in the initial steps of neurodegeneration. Here we review and discuss known contributors, including variants in genetic loci and aberrant protein activities, which modify cytoskeletal integrity, axonal transport and mitochondrial localization in ALS and have overlapping features with other neurodegenerative diseases. Additionally, we explore some emerging pathways that may contribute to this disruption in ALS.

Meta-Analysis of In-Vitro Bonding of Glass-Ionomer Restorative Materials to Primary Teeth.

Restoration of primary teeth is among the main clinical applications of glass-ionomer cements (GIC). The aim of the study was to review and summarize existing evidence of in vitro bond strength of glass-ionomer (GI) restoratives to enamel and dentin of primary teeth. A literature search was performed in PubMed/Medline, Scopus, Web of Science, Cochrane, and Google Scholar databases to identify studies published until April 2021. The search strategy was: ("glass") and ("ionomer") and ("primary" or "deciduous") and ("bond" or "tensile" or "shear"). Two researchers independently retrieved articles that reported on the bond strength of GIC to primary dentin and/or enamel. The meta-analysis was performed to compare the bond strength values of conventional (C) GIC and resin-modified (RM) GIC to different substrates. From 831 potentially eligible articles, 30 were selected for the full-text examination, and 7 were included in the analysis. Studies were rated at high (3), medium (3), and low (1) risk of bias. RM-GIC showed higher bond strength to primary enamel and dentin compared to the C-GIC. Meta-analysis of in vitro studies, evaluating bonding properties of GI restoratives to primary teeth, suggests the superior performance of RM-GIC. However, there is a lack of studies that examine the properties of novel GI formulations.

Crown formation times of deciduous teeth and age at death in neolithic newborns / Tiempos de formación de la corona de los dientes deciduos y edad al morir en neonatos neolíticos

SUMMARY: The aim of the present study was to investigate the possibility of estimating crown formation times of immature deciduous teeth and age at death in Neolithic newborns. In the Neolithic-Mesolithic transition, the health of the population deteriorated. Leaving the intrauterine environment for the newborn is the first obstacle in the process of adaptation and survival in the outside world. The fetus is protected by the mother's immune system and receives the necessary nutrients through the umbilical cord, but external factors indirectly affect its development. At birth deciduous teeth are not fully formed and are only partially mineralized. Variations in the rhythmic activity of ameloblasts and the secretion of the enamel matrix lead to the formation of incremental lines in the enamel. The sample consisted of unerupted deciduous teeth removed from the baby jaws from Neolithic archaeological graves, LepenskiVir Serbia. The skeletal age of the babies was from 38 to 40 gestational weeks. The daily enamel apposition rate was obtained for each tooth. The age of individuals was estimated using crown formation time. The average value of daily secretion rates for the primary teeth from the Neolithic age was 3.78 µm. There was no statistically significant difference in age at death determined by skeletal age assessment and crown formation time. Three babies were born preterm. The results of the present study show that the calculation of the time required for the formation of deciduous tooth enamel is applicable to archaeological samples of newborns.The age estimation using crown formation time together with the analysis of other anthropological parameters, can contribute to a more accurate determination of neonatal death in anthropological, archaeological and forensic contexts.

RESUMEN: El objetivo del estudio fue investigar la posibilidad de estimar el tiempo de formación de la corona de los dientes deciduos inmaduros y la edad, al momento de la muerte en neonatos neolíticos. Durante la transición Neolítico-Mesolítico, la salud de la población deterioró significativamente. Para el recién nacido dejar el medio intrauterino es el primer obstáculo en el proceso de adaptación y supervivencia en el mundo exterior. El feto está protegido por el sistema inmunológico de la madre y recibe los nutrientes necesarios a través del cordón umbilical, pero factores externos afectan indirectamente su desarrollo. Al nacer, los dientes deciduos no están completamente formados y solo están parcialmente mineralizados. Las variaciones en la actividad rítmica de los ameloblastos y la secreción de la matriz del esmalte conducen a la formación de líneas incrementales en el esmalte. La muestra consistió en dientes sin erupción extraídos de las mandíbulas de neonatos de tumbas arqueológicas neolíticas, LepenskiVir Serbia. La edad esquelética de los bebés fue de 38 a 40 semanas de gestación. Se obtuvo la tasa diaria de aposición de esmalte para cada diente. La edad de los individuos se estimó utilizando el tiempo de formación de la copa. El valor promedio de las tasas de secreción diaria para los dientes temporales del Neolítico fue de 3,78µm. No hubo diferencia estadísticamente significativa en la edad al momento de la muerte determinada por la evaluación de la edad esquelética y el tiempo de formación de la corona. Tres bebés nacieron prematuros. Los resultados del presente estudio muestran que el cálculo del tiempo requerido para la formación del esmalte dental deciduo es aplicable a muestras arqueológicas de recién nacidos. La estimación de la edad utilizando el tiempo de formación de la corona junto con el análisis de otros parámetros antropológicos, puede contribuir a una mayor determinación precisa de la muerte neonatal en contextos antropológicos, arqueológicos y forenses.

Using histological staining techniques to improve visualization and interpretability of tooth cementum annulation analysis / Uso de técnicas de tinción histológica para mejorar la visualización y la interpretación del análisis de la anulación del cemento dental

SUMMARY: The aim of the present study was to evaluate the effect of different staining techniques on applicability and accuracy of tooth cementum annulation (TCA) method. Nine decalcination techniques, 8 dehydration protocols and 8 different techniques were applied in 3 teeth from the persons of a known age. Black and white, and color images of histological sections were captured. An x- ray was taken of each tooth and they were photographed. Researchers were asked to observe both black/white and color images of histological sections. Researchers were divided into two groups. The first group analyzed histological images only, and the second group had photos of teeth and X-rays. In the first group of observers (without X ray) the differences in age estimation between real and observed age were significant for 2 younger patients, but not for the oldest patient, where the observed and real values matched. Of the 6 raters, the assesments of the last 3 (that used x-ray images together with histological sections) did not differ significantly from the real values. Extensive analysis and multiple repetitions performed in the present investigation revealed that the most optimal method of decalcification for TCA method was EDTA II for a period longer than 14 days at a section thickness of 2-3mm, while the most optimal protocol for dehydration was number IV. When it comes to staining, the most optimal staining protocol used for the cemental lines visualization and counting was Crocein Scarlet/Acid Fuchsin staining and Toluidine blue staining used at semithin section. Additional use of preexperimental evaluation employing x-ray of analyzed teeth decreased the errors of age estimation.

RESUMEN: El objetivo del presente estudio fue evaluar el efecto de diferentes técnicas de tinción sobre la aplicación y precisión del método de anulación de cemento dental (TCA). Se usaron nueve técnicas de descalcinación, 8 protocolos de deshidratación y 8 técnicas diferentes en 3 dientes de personas de edad conocida. Se capturaron imágenes en blanco y negro y en color de cortes histológicos. Se tomó una radiografía de cada diente y se fotografiaron. Los investigadores observaron las imágenes en blanco y negro y en color de las secciones histológicas. Los investigadores se dividieron en dos grupos; el primer grupo analizó solo imágenes histológicas y el segundo grupo tenía fotografías de los dientes y las radiografías. En el primer grupo de observadores (sin rayos X) las diferencias en la estimación de la edad entre la edad real y la edad observada fueron significativas para 2 pacientes más jóvenes, pero no para el paciente de mayor edad, donde los valores observados y reales coincidieron. De los 6 evaluadores, las valoraciones de los 3 últimos (que utilizaron imágenes de rayos X junto con cortes histológicos) no difirieron significativamente de los valores reales. El análisis exhaustivo y las múltiples repeticiones realizadas en la presente investigación revelaron que el método de descalcificación más óptimo para el método TCA fue EDTA II durante un período superior a 14 días con un grosor de sección de 2-3 mm, mientras que el protocolo óptimo para la deshidratación fue el número IV. En lo que respecta a la tinción, el protocolo de tinción más óptimo utilizado para la visualización y el recuento de las líneas de cemento fue la tinción con croceína escarlata / fucsina ácida y la tinción con azul de toluidina utilizada en la sección semifina. El uso adicional de la evaluación pre-experimental que emplea los rayos X de los dientes analizados disminuyó los errores de estimación de la edad.