Dental Pulp Chamber Research Articles

Systemic Statin Use and Pulp Chamber Calcification: A Pilot Retrospective Case Control Study Using Cone-Beam Computed Tomography.

This pilot retrospective case-control study questioned whether systemic statin use causes pulp calcification using cone-beam computed tomography images from the patients prescribed oral statins and comparing those of healthy individuals. CBCT scans of 54 patients, including 27 age- and sex-matched patients for the study and control groups, were analysed using Mimics Innovation Suite software. The study included patients using statins regularly for at least 1 year. Only intact teeth with opposing teeth were selected for the study group and matched with the control group. Dental crown and pulp chamber volumes were calculated and proportioned. The data were analysed with chi-square and Shapiro-Wilk tests to assess normal distribution, followed by Mann-Whitney U test if necessary. Statistical analysis showed no difference between the study and control groups (p = 0.505). Statin use duration did not cause statistically significant difference in terms of the reduction of pulp chamber volume (p = 0.141). Within the limitations of the study, systemic statin use did not cause dental pulp calcification. The results suggest, oral administration of the statin drugs is not an unfavourable condition for dental practice. Further studies with larger numbers of patients are needed to support this conclusion.

Cone-Beam Computed Tomography for the Evaluation of Dental Pulp Chamber Volume: Implications for Clinics and Teaching.

The dental pulp chamber volume is a fundamental measurement in the field of endodontics, but also in forensic sciences, teaching and training, or tissue engineering. This study evaluates the precision of cone-beam computed tomography (CBCT) in comparison with computed micro-tomography (micro-CT) in evaluating the pulp chamber volume of the upper central incisors ex vivo. The intra-operator and inter-operator errors were evaluated, and the results for the two techniques were compared with those of a T-test for paired samples. The intra-operator and inter-operator errors were >0.05, indicating adequate reproducibility in each operator and no significant differences between their measurements. On the other hand, no significant differences between the two measurement techniques were found. The present results demonstrate that CBCT is a precise, feasible, and reproducible technique for the evaluation of the dental pulp chamber volume ex vivo. The results provided with this method are useful for different medical domains but also for the teaching and training of undergraduate and postgraduate students. Furthermore, the findings of this study carry significant clinical implications, as the accurate assessment of the pulp chamber volume is critical in the diagnosis and treatment of various endodontic conditions. The ability of CBCT to provide reliable 3D dental anatomy measurements can enhance the planning of endodontic treatments by allowing for a better understanding of the internal tooth morphology. Additionally, the precision and reproducibility of CBCT in assessing the pulp chamber volume can contribute to improved clinical outcomes and reduced complications during endodontic procedures. These findings further support the increasingly vital role of CBCT in modern clinical practice and underscore its value as an indispensable tool in the field of dentistry.

Contribution of Anatomy in Forensic Age Evaluation: A Systematic Review.

The field of forensic anthropology is characterised by its ongoing development and growth. Forensic anatomy is a burgeoning discipline that focuses on the analysis and identification of both preserved and unpreserved human body parts, both in deceased individuals and the living. This subject plays a crucial role in establishing the four key factors of forensic anthropology, namely sex, age, race, and height. The objective of this research endeavour was to evaluate the significance of anatomical information in the process of forensic age estimation. The researchers established the inclusion criteria in accordance with the globally recognised Population, Intervention, Comparison, Outcome(PICOS) framework, as advised by the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) recommendations. The research included many methodologies in order to ascertain the age. Upon conducting a comprehensive review of the existing literature pertaining to anatomical knowledge in the field of forensic age estimate, we have identified many notable applications. These include the utilisation of various anatomical features such as the dental pulp chamber, fingerprints, acetabulum, sternal end of the fourth rib, as well as hand and wrist bones for the purpose of age estimation. It is important for anatomists and other forensic scientists to engage in collaborative efforts to facilitate the exchange of ideas and ensure thorough investigations. This cooperation is particularly crucial in areas where anatomical sciences play a significant role in forensic science and investigation. Nevertheless, in order to mitigate the potential for estimating error, it is still advisable to use a multi-factorial evaluation approach that involves examining many body areas.

Constitutive expression of spliced X-box binding protein 1 inhibits dentin formation in mice.

Upon endoplasmic reticulum (ER) stress, inositol-requiring enzyme 1 (IRE1) is activated, which subsequently converts an unspliced X-box binding protein 1 (XBP1U) mRNA to a spliced mRNA that encodes a potent XBP1S transcription factor. XBP1S is essential for relieving ER stress and secretory cell differentiation. We previously established Twist2-Cre;Xbp1 CS/+ mice that constitutively expressed XBP1S in the Twist2-expressing cells as well as in the cells derived from the Twist2-expressing cells. In this study, we analyzed the dental phenotype of Twist2-Cre;Xbp1 CS/+ mice. We first generated a mutant Xbp1s minigene that corresponds to the recombinant Xbp1 Δ26 allele (the Xbp1 CS allele that has undergone Cre-mediated recombination) and confirmed that the Xbp1s minigene expressed XBP1S that does not require IRE1α activation in vitro. Consistently, immunohistochemistry showed that XBP1S was constitutively expressed in the odontoblasts and other dental pulp cells in Twist2-Cre;Xbp1 CS/+ mice. Plain X-ray radiography and µCT analysis revealed that constitutive expression of XBP1S altered the dental pulp chamber roof- and floor-dentin formation, resulting in a significant reduction in dentin/cementum formation in Twist2-Cre;Xbp1 CS/+ mice, compared to age-matched Xbp1 CS/+ control mice. However, there is no significant difference in the density of dentin/cementum between these two groups of mice. Histologically, persistent expression of XBP1S caused a morphological change in odontoblasts in Twist2-Cre;Xbp1 CS/+ mice. Nevertheless, in situ hybridization and immunohistochemistry analyses showed that continuous expression of XBP1S had no apparent effects on the expression of the Dspp and Dmp1 genes. In conclusion, these results support that sustained production of XBP1S adversely affected odontoblast function and dentin formation.

Hydrogen Peroxide Diffusion through Dental Tissues-In Vitro Study.

Whitening products commonly utilize hydrogen peroxide (HP) as an active principle, which can penetrate dental tissues with potential side effects due to its low molecular weight. This study aimed to evaluate the HP diffusion of two in-office whitening products, namely 6% VivaStyle Paint On Plus (VS) and Opalescence Boost 40% (OP), in different tooth types. Additionally, the influence of the area of exposure, dental tissue thickness and pulp chamber volume was assessed. Each group consisted of eighteen intact anterior (A), premolar (PM) and molar (M) human teeth, and a positive pulpal pressure model was employed. The samples were analyzed using spectrophotometry, and results were expressed as the mean and 95% confidence interval. Statistical tests and linear regression models were appropriately applied at α = 5%. The total HP (µg) retrieved was as follows: VS-A, 1.333 [1.214, 1.452]; OP-A, 1.538 [1.457, 1.620]; VS-PM, 1.208 [1.123, 1.291]; OP-PM, 3.628 [3.401, 3.855]; VS-M, 2.560 [2.297, 2.823]; and OP-M, 4.197 [3.997, 4.396], with statistically significant differences in diffusion kinetics between whitening products for PM and M. Several HP concentrations attained a minimum cytotoxicity value of 2.22 µg/mL. The regression model shows that OP exposed the pulp chamber to 1.421 µg of HP more than that of VS. Different whitening products can cause cytotoxic HP concentrations in the pulp chamber, with a higher risk observed in molars.

Emerging Roles of YAP/TAZ in Tooth and Surrounding: from Development to Regeneration.

Yes associated protein (YAP) and transcriptional coactivator with PDZ-binding motif (TAZ) are ubiquitous transcriptional co-activators that control organ development, homeostasis, and tissue regeneration. Current in vivo evidence suggests that YAP/TAZ regulates enamel knot formation during murine tooth development, and is indispensable for dental progenitor cell renewal to support constant incisor growth. Being a critical sensor for cellular mechano-transduction, YAP/TAZ lays at the center of the complex molecular network that integrates mechanical cues from the dental pulp chamber and surrounding periodontal tissue into biochemical signals, dictating in vitro cell proliferation, differentiation, stemness maintenance, and migration of dental stem cells. Moreover, YAP/TAZ-mediated cell-microenvironment interactions also display essential regulatory roles during biomaterial-guided dental tissue repair and engineering in some animal models. Here, we review recent advances in YAP/TAZ functions in tooth development, dental pulp, and periodontal physiology, as well as dental tissue regeneration. We also highlight several promising strategies that harness YAP/TAZ activation for promoting dental tissue regeneration.

The Wisdom in Teeth: Neuronal Differentiation of Dental Pulp Cells.

Mesenchymal stem/stromal cells (MSCs) are found in almost all postnatal organs. Under appropriate environmental cues, multipotency enables MSCs to serve as progenitors for several lineage-specific, differentiated cell types. In vitro expansion and differentiation of MSCs give the opportunity to obtain hardly available somatic cells, such as neurons. The neurogenic potential of MSCs makes them a promising, autologous source to restore damaged tissue and as such, they have received much attention in the field of regenerative medicine. Several stem cell pool candidates have been studied thus far, but only a few of them showed neurogenic differentiation potential. Due to their embryonic ontology, stem cells residing in the stroma of the dental pulp chamber are an exciting source for in vitro neural cell differentiation. In this study, we review the key properties of dental pulp stem cells (DPSCs), with a particular focus on their neurogenic potential. Moreover, we summarize the various presently available methods used for neural differentiation of human DPSCs also emphasizing the difficulties in reproducibly high production of such cells. We postulate that because DPSCs are stem cells with very close ontology to neurogenic lineages, they may serve as excellent targets for neuronal differentiation in vitro and even for direct reprogramming.

Three-Dimensional Analysis of the Pulp Chamber and Coronal Tooth of Primary Molars: An In Vitro Study.

The anatomical variability of primary molars promotes research to improve clinical restorative activity, forensic identification, and industrial development. The objective was to study the morphology of the pulp chamber and its three-dimensional relationship with the external morphology of the tooth. A total of 151 deciduous molars were collected and subjected to three-dimensional imaging analysis in order to determine dental crown (CV) and pulp chamber (PV) volumes, their ratio (VR), pulp chamber roof (PCR) and pulp chamber floor (PCF) area, the furcation length (FD), and morphological peculiarities. The data were compared using the Kruskal–Wallis test in SPSS 26 software. The statistical analysis determined statistically significant differences between the four groups of molars for all variables. Great anatomical variability was observed, especially in the maxillary first molar, the volumes were higher in the maxillary second molar and the highest risk of furcal perforation was seen in the mandibular first molar. Additionally, mandibular second molars with six pulp horns, and some different density images compatible with accessory canals and pulp stones were found. Based on the findings of this investigation, we confirm the great anatomical variability that exists between the maxillary and mandibular molars.

Effects of Anti–Receptor Activator of Nuclear Factor Kappa B Ligand Antibody and Zoledronic Acid on Periapical Lesion Development in Mice

IntroductionAntiresorptive drugs are widely used to treat osteoporosis and other systemic bone diseases, although their efficacy for local bone resorption after localized inflammation has not been fully elucidated. We examined the effects of an anti–receptor activator of nuclear factor kappa B ligand (RANKL) antibody and the bisphosphonate zoledronic acid (ZOL) on periapical lesion (PL) development in mice. MethodsDental pulp of lower first molars in mice was removed, with the exposed dental pulp chambers left open to the oral environment to induce apical periodontitis. An anti-RANKL antibody or ZOL was intraperitoneally injected once per week until postoperative day 21, and then micro–computed tomographic imaging and histologic analyses were performed. ResultsPL enlargement was inhibited by both the anti-RANKL antibody and ZOL in a dose-dependent manner, and a reduction of inflammatory cell infiltration in apical tissues inhibited periapical bone resorption. The anti-RANKL antibody decreased the number of osteoclasts in periapical tissues, whereas ZOL suppressed periapical bone resorption with osteoclast numbers maintained. Although the administration of each of the antiresorptive drugs increased femoral bone mass, femoral bone mineral density in the PL group was lower compared with the sham-operated group. ConclusionsThese results suggest that an antiresorptive drug administered systemically is distributed to areas of local inflammation in the jaw can prevent PL development.

Radiographic Images of the Narrowing of Dental Pulp Chambers in Patients Receiving Long-term Steroid Therapy

Radiographic Images of the Narrowing of Dental Pulp Chambers in Patients Receiving Long-term Steroid Therapy

The dental pulp chamber evaluation by using cone-beam computed tomography

Background: Cone-beam computed tomographic (CBCT) imaging is a valuable tool in dental practice. It is widely used in endodontic treatment for the root canal morphology examination. Therefore, the purpose of this study was to use CBCT to calculate the volume of the pulp chamber at different tooth groups. Material and methods: This study conforms to protocols approved and in accordance with the ethics committee’s requirements, informed consent was obtained from each patient. Morphologic measurements of 120 maxillary and 120 mandibular molars (from 40 patients, aged 18–45 years) were included in this study. CBCT images were taken using a Kodak 9500 (Dental Systems, Carestream Health) operated at 90 kVp with a voxel size of 300 mm and a field of view of 90-50 mm. All scans were taken following the manufacturer’s recommendation protocol. According to the examination requirements, C-shaped roots, single-rooted molars, crowned teeth, and teeth with caries and/or restorations violating the pulp chamber were excluded. All measurements were taken on the coronal plane view. Results: In the present study, we used CBCT imaging to gather information regarding pulp chamber volume. With the scanned 3-dimensional images, we were able to clinically determine the pulp chamber parameters using a standardized and defined spatial approach. Conclusions: The data we collected here serve as a proof of principle for the analysis of dental landmarks before collecting stem cells. In this particular study, existing CBCT scans were used to provide useful information that can be used as a guide to determine the volume of the pulp chamber

Heat stress defense in dental pulp stem cells

Many dental treatment procedures can generate excessive heat within a short period of time. The heat on the tooth surface can be transduced to the dental pulp chamber and lead to alteration of dental pulp stem cells, vascular structures and sensory neurons within the chamber. It has been reported that elevation of temperature by 5.5°C in the pulp chamber caused 15% irreversible pulpal damage. However, the molecular changes of dental pulp cells post heat stress has not been characterized. In this study, we explored the changes in human dental pulp Stem Cells from the Exfoliated Deciduous teeth (SHED) after exposure to heat stress.Lysate of SHED cells were collected after exposure to heat stress at 42°C or normal culture temperature at 37°C. Western blots were used to examine the expression of Heat Shock Proteins (HSP). Human cytokine arrays (AAH‐CYT‐5, RayBiotech) were used to evaluate the changes in the expression of 80 human cytokines in SHED cells after heat stress.Short term exposure to heat stress induced phosphorylation of HSP27. Prolonged exposure further increased expression of other heat shock proteins, which have been reported to play an important role in cell protection under stress. In addition, transient exposure to heat stress caused differential expression of cytokines in SHED cells, suppressing expression of cytokines as IL‐1a and IL‐10, while increasing expression of glial cell derived neurotrophic factor (GDNF) and insulin‐like growth factor ‐1 (IGF‐1).The results showed HSPs and multiple cytokines play important role in detecting and responding to heat stress on the SHED cells. These factors could potentially protect SHED cells from thermal damage and promote stem cell regeneration post thermal damage.Support or Funding InformationFaculty Scholarly/Artistic Activities Award DDNA12‐013 University of the Pacific.

Mutant Dentin Sialophosphoprotein Causes Dentinogenesis Imperfecta

Dentin sialophosphoprotein (DSPP) is an extracellular matrix protein highly expressed by odontoblasts in teeth. DSPP mutations in humans may cause dentinogenesis imperfecta (DGI), an autosomal dominant dentin disorder. We recently generated a mouse model (named “DsppP19L/+ mice”) that expressed a mutant DSPP in which the proline residue at position 19 was replaced by a leucine residue. We found that the DsppP19L/+ and DsppP19L/P19L mice at a younger age displayed a tooth phenotype resembling human DGI type III characterized by enlarged dental pulp chambers, while the teeth of older DsppP19L/+ and DsppP19L/P19L mice had smaller dental pulp chambers mimicking DGI type II. The teeth of DsppP19L/+ and DsppP19L/P19L mice had a narrower pulp chamber roof predentin layer, thinner pulp chamber roof dentin, and thicker pulp chamber floor dentin. In addition, these mice also had increased enamel attrition, accompanied by excessive deposition of peritubular dentin. Immunohistochemistry, in situ hybridization, and real-time polymerase chain reaction analyses showed that the odontoblasts in both DsppP19L/+ and DsppP19L/P19L mice had reduced DSPP expression, compared to the wild-type mice. We also observed that the levels of DSPP expression were much higher in the roof-forming odontoblasts than in the floor-forming odontoblasts in the wild-type mice and mutant mice. Moreover, immunohistochemistry showed that while the immunostaining signals of dentin sialoprotein (N-terminal fragment of DSPP) were decreased in the dentin matrix, they were remarkably increased in the odontoblasts of the DsppP19L/+ and DsppP19L/P19L mice. Consistently, our in vitro studies showed that the secretion of the mutant DSPP was impaired and accumulated within endoplasmic reticulum. These findings suggest that the dental phenotypes of the mutant mice were associated with the intracellular retention of the mutant DSPP in the odontoblasts of the DSPP-mutant mice.

Visualization of the pulp chamber roof and residual dentin thickness by spectral-domain optical coherence tomography in vitro.

The aim of this study was to evaluate the ability of spectral-domain optical coherence tomography (SD-OCT) to display the roof of the pulp chamber and to estimate the residual dentin thickness (RDT) of the pulp complex. The roots of 20 extracted human molars were embedded in epoxy resin, and crowns were longitudinally sectioned in the mesial-distal direction, exposing the pulp chamber. The coronal part of the crown was removed up to an RDT to the pulp chamber roof of 2mm. Samples were imaged by SD-OCT from coronal view and by light microscopy (LM) in the sagittal plane. Using a microtome, dentin was subsequently removed in four levels from the occlusal aspect in steps of 250μm. At each level, RDT was documented and measured by both methods. The data were compared (Spearman's rho correlation coefficient, Wilcoxon signed-rank test). Using OCT, the roof of the pulp chamber was first displayed at a maximum RDT of 1.94mm. The minimal RDT that could be imaged by OCT was 0.06mm. Values from both methods were strongly correlated (r, 0.83-0.95; pi ≤ 0.05) and differed significantly for large RDTs (dentin levels 1, 2; pi< 0.05) but not for small RDTs (levels 3, 4; pi ≥ 0.226). The roof of the dental pulp chamber could be already visualized by SD-OCT with a RDT of 1.94mm. Therefore, the method could be a useful diagnostic tool during the preparation of deep dentin cavities and might help to preserve the integrity of the pulp chamber.

Thermal Scanning of Dental Pulp Chamber by Thermocouple System and Infrared Camera during Photo Curing of Resin Composites.

Introduction:Due to thermal hazard during composite restorations, this study was designed to scan the pulp temperature by thermocouple and infrared camera during photo polymerizing different composites. Methods and Materials:A mesio-occlso-distal (MOD) cavity was prepared in an extracted tooth and the K-type thermocouple was fixed in its pulp chamber. Subsequently, 1 mm increment of each composites were inserted (four composite types were incorporated) and photo polymerized employing either LED or QTH systems for 60 sec while the temperature was recorded with 10 sec intervals. Ultimately, the same tooth was hemisected bucco-lingually and the amalgam was removed. The same composite curing procedure was repeated while the thermogram was recorded using an infrared camera. Thereafter, the data was analyzed by repeated measured ANOVA followed by Tukey’s HSD Post Hoc test for multiple comparisons (α=0.05). Results:The pulp temperature was significantly increased (repeated measures) during photo polymerization (P=0.000) while there was no significant difference among the results recorded by thermocouple comparing to infrared camera (P>0.05). Moreover, different composite materials and LCUs lead to similar outcomes (P>0.05). Conclusion:Although various composites have significant different chemical compositions, they lead to similar pulp thermal changes. Moreover, both the infrared camera and the thermocouple would record parallel results of dental pulp temperature.

Al-Awadi-Raas-Rothschild syndrome with dental anomalies and a novel WNT7A mutation

Al-Awadi-Raas-Rothschild syndrome (AARRS; OMIM 276820) is a very rare autosomal recessive limb malformation syndrome caused by WNT7A mutations. AARRS is characterized by various degrees of limb aplasia and hypoplasia. Normal intelligence and malformations of urogenital system are frequent findings. Complete loss of WNT7A function has been shown to cause AARRS, however, its partial loss leads to the milder malformation, Fuhrmann syndrome. An Indian boy affected with AARRS is reported. A novel homozygous base substitution mutation c.550A > C (p.Asn184Asp) is identified in the patient. Parents were heterozygous for the mutation. In addition to the typical features of AARRS, the patient had agenesis of the mandibular left deciduous lateral incisor. The heterozygous parents had microdontia of the maxillary left permanent third molar and taurodontism (enlarged dental pulp chamber at the expense of root) in a number of their permanent molars. Whole exome sequencing of the patient and his parents ruled out mutations in 11 known hypodontia-associated genes including WNT10A, MSX1, EDA, EDAR, EDARADD, PAX9, AXIN2, GREM2, NEMO, KRT17, and TFAP2B. In situ hybridization during tooth development showed Wnt7a expression in wild-type tooth epithelium at E14.5. All lines of evidence suggest that WNT7A has important role in tooth development and its mutation may lead to tooth agenesis, microdontia, and taurodontism. Oral examination of patients with AARRS and Fuhrmann syndromes is highly recommended.

Interface Between MTA and Dental Bonding Agents: Scanning Electron Microscope Evaluation.

Aims and Objectives:Nowadays, the material that offers the best sealing characteristic in the field of endodontic treatment is the mineral trioxide aggregate (MTA), nevertheless, this material necessities an adhesive bonding agent to perfectly join to the dental surface. The aim of this study was to analyze using a scanning electron microscope (SEM) the possible microgap between the adhesive, MTA, and the dental surface.Material and Methods:Fourteen extracted molars were divided into two groups – group A was prepared with MTA-component adhesive and group B was prepared with MTA and composite dual etching. The observations were carried out with a SEM Phenom G2 Pro mode S.E.I. JMP® software was used for statistical analysis, and a t-test was used for evaluating the difference between the two groups.Results:The gap of the areas at higher magnification (1000×) with a size greater than 5 microns in width and 20 microns in length were considered significant, and only group A recorded significant data.Conclusions:The SEM analysis performed in the group A with interposition of adhesive and flow between the dental pulp chamber and MTA demonstrates the presence of a marginal gap of considerable amplitude in the all of the samples investigated.

Oral Symptoms and Bone Observations in Odonto-Hypophosphatasia

Hypophosphatasia (HPP) is an inherited skeletal disease caused by mutation of the gene encoding tissue non-specific alkaline phosphatase (TNSALP). Odonto-HPP is well known as the mildest of HPP. The manifestations involve only the teeth, such as premature primary teeth exfoliation caused by reduction of alveolar bone, enlarged dental pulp chamber, and dental defects. We report a case of a 9-years-old boy who developed HPP. He was observed from the primary dentition to the mixed dentition period. At initial presentation at our hospital, he had multiple premature exfoliation of primary teeth and reduction of the alveolar bone. HPP was suspected due to the low level of ALP activity in serum, his oral manifestation, and dental history. He was referred to a physician for the final diagnosis. Therefore his compound heterozygote mutations, c.1559 delT (T/delT) and c.407G > A (G/A), were found in TNSALP and he diagnosed with odonto-HPP. Even though these mutations were reported as being involved in odonto-HPP, his mineral densities tended to be lower than that of his age. It is therefore necessary to investigate the bone mineralization level in odonto-HPP without other bone symptoms. Moreover, ongoing enzyme-replacement therapy in odonto-HPP might improve dental abnormality and bone disorders.

Prevalence of and relationship between pulp and renal stones: A radiographic study

AimThe aim of the present study was to determine the prevalence of and the relationship between pulp and renal in affected patients and in healthy adults. Materials and methodsA total of 240 patients participated in the study. Group A consisted of 120 patients who had renal calculi and Group B had 120 randomly selected controls for the study. The periapical radiographs for all patients were evaluated for the presence or absence of the narrowing of dental pulp chambers and pulp canals. The radiographs were also evaluated to determine the presence or absence of pulp stones. The results were compared and analyzed using the Chi-square test (p<0.001). ResultsA total of 164 patients had pulp narrowing and 112 patients had pulp stones, which included 55 controls and 57 renal calculi patients. There was no statistical correlation between pulp narrowing and renal stones (p>0.001) and also between pulp stones and renal stones (p>0.001). ConclusionHowever, there was no significant correlation between the presence of pulp stones and renal stones, and the incidental findings of pulp stones on periapical radiographs can provide useful information in the early diagnosis of the systemic calcifications.

Evaluation of DNA typing as a positive identification method for soft and hard tissues immersed in strong acids.

Identification of human remains can be hindered by several factors (e.g., traumatic mutilation, carbonization or decomposition). Moreover, in some criminal cases, offenders may purposely adopt various expedients to thwart the victim's identification, including the dissolution of body tissues by the use of corrosive reagents, as repeatedly reported in the past for Mafia-related murders. By means of an animal model, namely porcine samples, we evaluated standard DNA typing as a method for identifying soft (muscle) and hard (bone and teeth) tissues immersed in strong acids (hydrochloric, nitric and sulfuric acid) or in mixtures of acids (aqua regia). Samples were tested at different time intervals, ranging between 2 and 6h (soft tissues) and 2-28 days (hard tissues). It was shown that, in every type of acid, complete degradation of the DNA extracted from soft tissues preceded tissue dissolution and could be observed within 4h of immersion. Conversely, high molecular weight DNA amenable to STR analysis could be isolated from hard tissues as long as cortical bone fragments were still present (28 days for sulfuric acid, 7 days for nitric acid, 2 days for hydrochloric acid and aqua regia), or the integrity of the dental pulp chamber was preserved (7 days, in sulfuric acid only). The results indicate that DNA profiling of acid-treated body parts (in particular, cortical bone) is still feasible at advanced stages of corrosion, even when the morphological methods used in forensic anthropology and odontology can no longer be applied for identification purposes.