Proteomic signatures of equine dental tooth tissues in ageing and disease.

Trace elements play a key role in dental tissue development, as dental hard tissues accumulate both essential and toxic trace elements during mineralization. Characterization of the spatial accumulation pattern of trace elements may provide insight into exposure to toxic elements over time and to the nature of disease processes affecting the hard dental tissues. Here, we present the first report of the use of laser ablation-inductively coupled plasma-mass spectrometry (LA-ICP-MS) to map the microspatial distribution of multiple trace elements, essential and toxic, across feline dental hard tissues. Eleven teeth were extracted from 8 cats. Nine teeth were from 7 cats diagnosed with idiopathic tooth resorption on intraoral radiographs prior to extraction. Two teeth were included from a cadaver that had no signs of tooth resorption on intraoral radiographs. The normal dental tissue was analyzed from each sample using LA-ICP-MS to map the microspatial distribution of essential and toxic trace elements across feline enamel, dentin, and cementum. Results showed a higher accumulation of barium and strontium in coronal dentin as compared to root dentin. The timing of the accumulation mirrors nursing timelines seen in teeth from human and non-human primates, consistent with barium and strontium being sourced from maternal milk. Results also showed a higher uptake of lead in the coronal dentin, suggesting this lead exposure was likely passed from mother to offspring. This work characterizes a baseline for elemental distribution in feline teeth linked to early life exposure to toxic elements such as lead and provides a framework for future studies investigating long-term environmental exposures to trace elements, essential and toxic, and their involvement in feline systemic and dental diseases.

BackgroundPost-mortem toxicology constantly deals with the research of reliable alternative matrices to be applied in case of highly damaged corpses (such us carbonized, skeletonized, human remains, etc.). Teeth represent a promising alternative matrix since dental tissues are endowed by different features, resistance and stability after death. ScopeSince scant literature reported on the pharmacokinetics and mechanism of incorporation of xenobiotics into dental tissues, this pilot research aims to investigate whether in the pulp can be detected the same substances found in blood in drug related death cases. Secondly, the study is addressed to disclose the possible deposit of drugs in dental hard tissues (dentine and/or enamel), thus contributing to reconstruct the drug abuse history (timing, e.g.). Materials and methodsThe study experimented with a novel method to separately analyse dental enamel, dentin, and pulp, applied to 10 teeth collected during autopsies of drug-related deaths along with blood and hair samples for classic toxicological analyses. Each tooth was prepared by “pulverization technique” and then analysed by gas chromatography paired with mass spectrometry (GC-MS) and ultra high performance liquid chromatography coupled to high resolution mass spectrometry (UHPLC/HR-MS) for searching cocaine, opiates, and metabolites. The results were then compared with those obtained from blood and hair samples. ResultsPreliminary results demonstrated that teeth differ from any other classic matrix (blood and hairs) since the qualitative correspondence of the detected substances between pulp and blood as well as dental hard tissues and hair suggests that they can be useful in post-mortem evaluation as a unique matrix for both acute and chronic assumptions of drugs. The mechanism of accumulation of substances in mineralized dental tissues emerged the most significant result, being influenced by the type of molecule and the method of assumption. The main limitation of this study is the limited availability of the sample and the absence of anamnestic information of the time, rates and method of drug assumption during life. Further research is necessary to systematically investigate the distribution of different substances within the different tissues of the tooth.

Objective: To describe how diverse dental tissues (enamel, dentin and cement) and four dental materials (amalgam, composite, glass ionomer and zinc oxide modified cement) react after high temperature exposure. Dental tissue and dental materials can be used to identify human remains.Methods: An experimental in vitro study was carried out to determine physical macroscopic and microscopic changes in dental tissues and four dental materials, using 199 extracted teeth after high temperature exposition. Fire exposure at diverse temperatures ranking (200ºC, 400ºC, 600ºC, 800ºC, 1000ºC, 1200ºC) were performed in dental materials and tissues to determine their physical stability.Results: Dental tissues and the four dental materials evaluated had high resistance to high temperatures without apparent variation in their physical structure. These features in both dental tissues and dental materials could be useful in forensic identification. However, small changes occurred after temperaure exposure such as dimensional expansion, presence of fissures, cracks, fractures and color change on dental tissues and dental materials.Conclusions: This study determines that hard dental tissues and dental materials were able to resist high temperatures without high physical variants that may facilitate forensic identification of human remains after fire exposure.

Localization of Type V Collagen with Monoclonal Antibodies in Developing Dental and Peridental Tissues of the Rat and Hamster

The great interest in the field of laser applications in dentistry provokes the question, if all these new techniques may really fulfill advantages, which are expected after initial in-vitro studies. Whereas laser surgery of soft oral tissues has been developed to a standard method, laser treatment of dental hard tissues and the bone are attended with many unsolved problems. Different laser types, especially pulsed lasers in a wide spectrum of wavelengths have been proofed for dental use. Today neither the excimer lasers, emitting in the far uv-range from 193 to 351 nm, nor the mid-infrared lasers like Nd:YAG (1,064 μm), Ho:YAG (2,1 μm) and Er:YAG (2,96 μm) or the C0₂-laser (10,6 μm) show mechanism of interaction more carefully and faster than a preparation of teeth with diamond drillers. The laser type with the most precise and considerate treatment effects in the moment is the short pulsed (15 ns) ArF-excimer laser with a wavelength of 193 nm. However this laser type has not yet the effectivity of mechanical instruments and it needs a mirror system to deliver the radiation. Histological results point out, that this laser shows no significant pathological alterations in the adjacent tissues. Another interesting excimer laser, filled with XeCI and emitting at a wavelength of 308 nm has the advantage to be good to deliver through quartz fibers. A little more thermal influence is to be seen according to the longer wavelength. Yet the energy density, necessary to cut dental hard tissues will not be reached with the laser systems available now. Both the pulsed Er:YAG- (2,94 μm, pulse duration 250 s) and the Ho:YAG -laser (2,1 μm, pulse duration 250 μs) have an effective coupling of the laser energy to hydrogeneous tissues, but they do not work sufficient on healthy enamel and dentine. The influence to adjacent healthy tissue is not tolerable, especially in regard of the thermal damage dentine and pulp tissues. Moreover, like the 193 nm ArF-excimer laser radiation the Er:YAG-laser radiation could also only be delivered via mirror systems, while the radiation of the Ho:YAG-Laser can be well transmitted through quartz fibers. The energy of the well known and in other medical disciplines often used Nd:YAG - laser (1,064 μm, pulse duration 150 us) laser can be transmitted through fiber systems without problems, but this laser has not the effectivity to work sufficient on healthy hard dental tissues due to the high transmission in mineralized dental tissues. The thermal injuries of this laser type are not tolerable. The short pulsed TEA-C0₂-laser (9,6 and 10,6 μm, pulse duration 200-300 ns), which has an excellent coupling not only to the hydrogeneous tissues but also to the mineralized tissues could be an alternative system to prepare dental tissues. The greatest disadvantage of this system is the noneffective delivery of the light energy through flexible fiber systems, which are still in development. Another good chance perhaps will have the q-switched Neodym, Erbium and Holmium:- YAG lasers with pulse durations of about some hundred ns. Both, possible thermal influences and possible disruptive effects should be small enough to let the adjacent tissues undamaged.

BackgroundThe development of ultrasound for use in dental tissues is hampered by the complex, multilayered nature of the teeth. The purpose of this preliminary study was to obtain the phase and group velocities associated with several directions of ultrasonic wave propagation in relation to the tooth structure, which would then lead to the determination of the elastic constants in dental hard tissue. Knowledge of these elastic constants can be used to feed back into numerical models (such as finite element) in order to simulate/predict ultrasonic wave propagation and behavior in the teeth. This will help to optimize ultrasonic protocols as potential noninvasive therapeutic tools for novel dental regenerative therapies.MethodsAn extracted human second molar was used to determine time-of-flight information from A-scan signatures obtained at various angles of inclination and rotation using a scanning acoustic microscope at 10 MHz. Phase and group velocities and associated slowness curves were calculated in order to determine the independent elastic constants in the human teeth.ResultsResults show that as the tooth was inclined at three azimuthal angles (Θin = 0°, 15°, and 30°) and rotated from Φin = 0° to 360° in order to cover the whole perimeter of the tooth, slowness curves constructed from the computed phase and group velocities versus angle of rotation confirm the inhomogeneous and anisotropic nature of the tooth as indicated by the nonuniform appearance of uneven circular shape patterns of the measurements when compared to those produced in a control isotropic fused quartz sample.ConclusionsThis study demonstrates that phase and group velocities of ultrasound as determined by acoustic microscopy change and are dependent on the direction of the tooth structure. Thus, these results confirm that the tooth is indeed a multilayered anisotropic structure underscoring that there is no single elastic constant sufficient to represent the complex structure of the tooth. The findings underline the importance to take into account these crucial characteristics in order to develop and optimize therapeutic as well as diagnostic applications of ultrasound in dental tissue repair, and further studies are warranted to analyze ultrasound transmission at various frequencies and intensities in different teeth to develop appropriate models for ultrasound biophysical behavior in dental tissues.

As people live longer, there is an increasing need for hard tissue regeneration and whole-tooth regeneration. Despite the advancements in the field of medicine, the field of regenerative dentistry is still challenging due to the complexity of dental hard tissues. Cross-disciplinary collaboration among material scientists, cellular biologists, and odontologists aimed at developing strategies and uncovering solutions related to dental tissue regeneration. Methodology. A search of the literature was done for pertinent research. Consistent with the Preferred Reporting Items for Systematic Review and Meta-Analysis (PRISMA) 2020 Statement, the electronic databases looked at were PubMed, Science Direct, Scopus, and Google Scholar, with the keyword search "hard dental tissue regeneration." Database analysis yielded a total of 476 articles. 222 duplicate articles have been removed in total. Articles that have no connection to the directed regeneration of hard dental tissue were disregarded. The review concluded with the inclusion of four studies that were relevant to our research objective. Current molecular signaling network investigations and novel viewpoints on cellular heterogeneity have made advancements in understanding of the kinetics of dental hard tissue regeneration possible. Here, we outline the fundamentals of stem hard dental tissue maintenance, regeneration, and repair, as well as recent advancements in the field of hard tissue regeneration. These intriguing findings help establish a framework that will eventually enable basic research findings to be utilized towards oral health-improving medicines.

Extracellular matrix (ECM) homeostasis is essential for normal tissue function, and its disruption by iatrogenic injury, trauma, or disease results in fibrosis. Skin ECM homeostasis is maintained by a complex process that involves an integration of cytokine and environmental mediators. However, it is unclear, in both normal and disease states, how these multifactorial processes converge to shift ECM homeostasis towards accumulation or degradation. Here we show a consistent downregulation in fatty acid oxidation (FAO) and upregulation of glycolysis in fibrotic skin and in normal skin with abundant ECM. Perturbation of glycolysis and FAO pathway enzymes reveals their reciprocal effects in ECM upregulation and downregulation, respectively. Increasing peroxisome proliferator-activated receptor (PPAR) signalling, an inducer of the FAO pathway, generates a catabolic fibroblast phenotype characterised by inhibition of ECM transcription and enhanced ECM internalization and lysosomal degradation. In contrast, suppression of glycolysis inhibits ECM gene transcription and protein levels, independently of an intact FAO pathway or PPAR signalling. Moreover, we show that CD36, a multifunctional fatty acid transporter, connects the metabolic state of fibroblasts with their capacity for ECM regulation, as internalization and degradation of collagen-1 is abrogated in fibroblasts lacking CD36. Finally, restoring FAO and upregulating CD36 reduces ECM accumulation in murine skin fibrosis. These findings indicate that metabolic perturbation of ECM homeostasis may have broad implications for therapies aimed at ECM regulation, such as fibrosis, regenerative medicine, and ageing.

Dental disease in hill sheep: An abattoir survey

The goal of the present study correlates dental hard tissue mineralization, mucosal pathologies in the oral cavity and different degrees of intestinal dysbiosis. the study examined two groups: the study group (Group I) included 229 children and adolescents aged 1-17 (mean age 5±1years) with oral pathologies (caries, acute or chronic candidiasis) and confirmed dysbiosis of varying severity and stages as well. Group II (the Control Group) was composed of 50 patients aged 1 - 16 (mean age 5±1years) with oral pathologies but with no detected changes in gastrointestinal (GI) flora. Dental caries were examined by DMFT-index; the extent of dental hard tissue mineralization by vital staining (2% methylene blue) and cases of oral candidiasis was investigated by taking cultures from mucosal plaques. on the basis of the research outcomes the correlation between the different degrees of GI dysbiosis and dental hard tissue mineralization with pathologic expressions in the oral cavity was found. Group I was divided into two subgroups: in the first subgroup that suffered from mild dysbiosis (I and II degree) moderate dental caries was revealed, whereas in the second subgroup with III and IV degree of dysbiosis-high levels of dental caries was detected. In Group II (no GI flora disturbances), the dental hard tissue demineralization indicator was minimal; in children aged 1-3 years the incidence and prevalence of caries were low and increased with age, reaching higher values during puberty (11-16 years). It may be concluded that dysbiosis of GI microflora influences on a degree of dental hard tissue demineralization, which in turn may predispose to the formation of dental caries.

To introduce a methodology designed to simultaneously visualize dental ultrastructures, including cellular and soft tissue components, by utilizing phosphotungstic acid (PTA) as a contrast-enhancement agent. Sound third molars were collected from healthy human adults and fixed in 4% buffered paraformaldehyde. To evaluate the impact of PTA in concentrations of 0.3%, 0.7% and 1% on dental soft and hard tissues for CT imaging, cementum and dentine-pulp sections were cut, dehydrated and stained with immersion periods of 12, 24h, 2days or 5days. The samples were scanned in a high-resolution nano-CT device using pixel sizes down to 0.5µm to examine both the cementum and pulpal regions. Dental cementum and periodontium as well as odontoblasts and predentine were made visible through PTA staining in high-resolution three-dimensional nano-CT scans. Different segments of the tooth required different staining protocols. The thickness of the cementum could be computed over the length of the tooth once it was made visible by the PTA-enhanced contrast, and the attached soft tissue components of the interior of the tooth could be shown on the dentine-pulp interface in greater detail. Three-dimensional illustrations allowed a histology-like visualization of the sections in all orientations with a single scan and easy sample preparation. The segmentation of the sigmoidal dentinal tubules and the surrounding dentine allowed a three-dimensional investigation and quantitative of the dentine composition, such as the tubular lumen or the ratio of the tubular lumen area to the dentinal surface. The staining protocol made it possible to visualize hard tissues along with cellular layers and soft tissues in teeth using a laboratory-based nano-CT technique. The protocol depended on both tissue type and size. This methodology offers enhanced possibilities for the concomitant visualization of soft and hard dental tissues.

The estimation of time of death or the determination of the postmortem interval (PMI) is one of the most important issues in forensic medicine and odontology. However, evaluation of bone and dental hard tissues in PMI could be challenging due to the lack of objective methods with high accuracy. In this respect, micro-CT analysis which has not been used for postmortem evaluation would be beneficial in evaluating hard tissues such as bones and teeth. Therefore, the aim of the present study was to determine the alterations in the hard dental tissue, mineral density of enamel, and the surface abrasion of hard dental tissues of rats in the PMI period with a relatively novel method, micro-CT. The present study included 60 female Wistar rats which were divided into six study groups. The rats were sacrificed at the baseline and were left into nature putrefaction process. The study groups were created based on the PMI period as week-0, week-1, week-2, week-4, week-8, and week-12, which included 10 rats in each group. All hemi-mandibles were collected in the determined timelines and the micro-CT analysis was carried out on each group. Mineral density of enamel and the surface abrasion of hard dental tissues were determined. The enamel and cement thickness remained the same in the examined PMI periods. Mineral density of the enamel tissues were also similar until the 8th week but the decrease was significant at 12th week (2.313gHApcm3). Surface abrasion of the dental tissues on weeks 4, 8, and 12 were 0.006, 0.024, and 0.024mm, respectively. The present study indicated that surface abrasion and enamel mineral density evaluation via micro-CT can be considered as objective and precise parameters in PMI evaluation in forensic medicine and odontology.

Currently, the issue of increased dentin sensitivity after operations for hard dental tissue preparation during the therapeutic treatment of the carious process, as well as during orthopedic treatment of various types with non-removable dentures, has retained its relevance. Pain management is the prevention of postoperative complication development and ensures high-quality treatment results. Aim. The purpose of the literature review in this paper was to draw specialists’ attention to the use of low-solvent adhesive systems of various types to manage postoperative sensitivity of hard dental tissues. Materials and methods. More than 30 literature sources focused on studying action mechanisms of known generations of dental adhesive systems and their effects on changes in postoperative sensitivity of dental hard tissues as a result of a high-quality protective film formation on the dentin surface have been analyzed. Results. The use of various adhesive systems to reduce postoperative sensitivity of soft tissues with the formation of a high-quality protective film on the dentin surface that seals the dentinal tubules is of great importance for obtaining qualitative treatment results and increasing its effectiveness. However, various reactive substances (solvents, acetone), which are components of most adhesives, have a strong impact on their quality and significantly reduce the indications for their use. Conclusions. Thus, studying the action mechanisms of adhesives on dental tissues, searching for the formulation of new adhesive systems with a lower solvent content would expand the indications for their use as therapeutic and prophylactic drugs to reduce pain sensitivity of hard dental tissues after preparation.

Most autoimmune diseases are multifactorial diseases and are caused by the immunological reaction against a number of autoantigens. Key for understanding autoimmune pathologies is the knowledge of the targeted autoantigens, both initially and during disease progression. We present an approach for autoantigen identification based on isolation of intact autoantibody-antigen complexes from body fluids. After organic precipitation of high molecular weight proteins and free immunoglobulins, released autoantigens were identified by quantitative label-free liquid chromatography mass spectrometry. We confirmed feasibility of target enrichment and identification from highly complex body fluid proteomes by spiking of a predefined antibody-antigen complex at low level of abundance. As a proof of principle, we studied the blinding disease autoimmune uveitis, which is caused by autoreactive T-cells attacking the inner eye and is accompanied by autoantibodies. We identified three novel autoantigens in the spontaneous animal model equine recurrent uveitis (secreted acidic phosphoprotein osteopontin, extracellular matrix protein 1, and metalloproteinase inhibitor 2) and confirmed the presence of the corresponding autoantibodies in 15-25% of patient samples by enzyme-linked immunosorbent assay. Thus, this workflow led to the identification of novel autoantigens in autoimmune uveitis and may provide a versatile and useful tool to identify autoantigens in other autoimmune diseases in the future.

To evaluate the correlation between the volume of dental hard tissues removed and the fracture resistance of mandibular molar teeth with ultraconservative (UltraAC) or traditional (TradAC) access cavity preparations. Sixty recently extracted and intact mandibular first molars were scanned in a microcomputed tomographic (micro-CT) device, anatomically matched and assigned at random to 2 groups (n=30), according to the access cavity type: UltraAC or TradAC. After access preparation, mesial and distal canals were prepared using Reciproc instruments and a new scan was taken. The volumes of pulp chamber and dental hard tissues in each specimen were measured before and after the experimental procedures, and the percentage volume reduction of hard tissues calculated for the entire tooth and for the crown separately. Teeth were then root filled, restored and subjected to fracture resistance tests in an universal testing machine. The force required to fracture was recorded in Newtons (N). The adherence of the variables to a Gaussian curve was verified using a Shapiro-Wilk test. Skewed data were analysed with nonparametric Wilcoxon signed rank or Spearman correlation tests, whilst the normally distributed data were analysed with a dependent samples Student t-test. Level of significance was set at 5%. Pre-operatively, no difference was observed between groups regarding the volume of pulp canal space or dental hard tissues (P>0.05). After access preparation, volumes of the removed hard tissues from the entire tooth and from the crown were significantly greater in teeth prepared with TradAC than with UltraAC (P<0.05). No significant difference was observed in the fracture resistance values between TradAC (902.9±347.8N) and UltraAC (948.7±405.7N) groups (P=0.975). The Spearman test was unable to identify a correlation between fracture resistance and percentage volume of removed dental hard tissues in the entire root (P=0.525, r=-0.084) or only in the crown (P=0.152, r=-0.187). The volume of hard dental tissues removed, although greater in teeth with TradAC compared to UltraAC, did not correlate with the fracture resistance results, indicating that a minimally invasive access cavity did not increase the resistance of restored mandibular first molars to fracture.