Effect of Heat on the Fluorescence Properties of Tooth‐Colored Restorative Materials and Their Forensic Implications

During antemortem and postmortem comparison of dental records of carbonized victims, restorations may be part of such records. The objective of this study was to evaluate the effect of heat on the fluorescence behavior of contemporary tooth‐colored restorative materials and natural tooth structure when subjected to range of temperatures, using illumination with 405 nm wavelength light. A total of 132 human extracted teeth restored with tooth‐colored restorative materials were exposed to heat (200, 500, 900, 1200°C) in an oven for 30 min. Samples were imaged before and after heat treatment. All tooth‐colored restorative materials underwent changes in color and in fluorescence properties, at each of the temperatures used. Resin‐based restorative materials still fluoresced at 200°C, and at 500°C underwent major color changes due to volatilization of resin. Materials containing inorganic fluorophores still fluoresced at 900°C, while at 1200°C, none of the materials tested in this study showed any fluorescence.     

Detection and classification of composite resins in incinerated teeth for forensic purposes

The great demand for esthetic restorations has resulted in placement of large numbers of composite resin fillings. The popularity of these materials is reflected in the quantity and variety of resin brands currently on the market. The ability to distinguish resin brands can aid in positive identification of burn victims, assuming that appropriate dental records exist. Scanning electron microscopy/energy dispersive X-ray spectroscopy (SEM/EDS) was used to characterize the structure and composition of 10 modern resins. The structure of each resin was unique to manufacturer, and elemental analysis allowed separation into distinct groups. These 10 resins were also placed in extracted teeth and incinerated at 900 degrees C for 30 min, simulating near cremation conditions. The resins were identifiable by SEM/EDS after incineration, and the elemental composition remained almost unchanged. The data produced are immediately useful for resin identification in forensics, and comparative analysis can be readily performed using standard equipment. This work represents the initial stage of database generation.     

Identification of incinerated root canal filling materials after exposure to high heat incineration

With the increase in global terrorism there is a higher probability of having to identify victims of incineration events secondary to incendiary explosive devices. The victims of incineration events challenge forensic odontologists when coronal restorations are no longer present to compile postmortem data. With 40 million root canals being completed annually in the United States, a very large pool of antemortem data is available to the forensic odontologist to make positive identifications. When complete and thorough dental records exist, individuals that have undergone surgical and nonsurgical root canal therapy may have materials present in the canal that may aid in identification. This study provides elemental fingerprints of root canal obturation materials to be utilized as a forensic identification aid. This study used scanning electron microscopy/energy dispersive X-ray spectroscopy (SEM/EDS) to assess the elemental composition of materials before and after high temperature incineration. Sixteen endodontic materials were analyzed pre-incineration and placed in extracted teeth. The filled teeth were subjected to incineration at 900 degrees C for 30 min to simulate incineration events or cremation. Incinerated materials were radiographed and re-analyzed to determine if they retained their original elemental composition. Endodontic sealers, gutta percha, root-end filling materials, silver points, and separated files were distinguishable in the canal and traceable after incineration. The authors present a fingerprint of the endodontic obturation materials that are capable of withstanding high heat incineration to be used as an aid for postmortem identification. This work represents the initial stage of database generation for root canal filling materials for use as an aid in forensic identification.     

Determining Volumetric Shrinkage Trends of Burnt Bone Using Micro-CT

Understanding the degree and pattern of shrinkage undergone by bone when subjected to heating is crucial to accurately deduce a biological profile from incinerated remains. X-ray microtomography (micro-CT) enables a nondestructive insight into hard tissue structural changes, while allowing for an accurate documentation of volumetric and trabecular shrinkage. Sheep ribs were experimentally burned at temperatures between 400 and 1000°C in 100°C increments and their volumetric shrinkage was calculated. Observed shrinkage ranged from 14.0% at 400°C to 45.5% at 1000°C. Bones burned at temperatures up to 600°C showed no significant difference, whereas the 700 and 800°C samples exhibited higher shrinkage. Bones burnt at 900 and 1000°C showed significantly higher shrinkage than the other temperature groups. Findings signify the potential of micro-CT in research on the effects of factors such as diagenesis or burning on the bone density, morphology and microarchitecture.     

Thermal Modifications of Root Transparency and Implications for Aging: A Pilot Study

Root transparency has proven to be related to age and has been considered by different odontological methods for age estimation. Very little is known concerning possible variations of root transparency with heat, although the applicability of the method to burnt remains depends on the possible modifications of this specific variable. This pilot study presents the results of an experiment performed on 105 teeth obtained from dental patients and autopsy material, heated in an industrial oven at 50°C, 100°C, 150°C and 200°C. Root transparency was measured before and after the charring experiment. The heating process proved to radically modify root transparency, which decreased in 20% of samples at 50°C, in 34.6% at 100°C, in 50% at 150°C, in 77% at 200°C. The overall correlation index (CI) between decrease in root transparency and increase in temperature amounted to 0.96. These results show that heat may modify root transparency and suggest caution in using methods based on root transparency for age estimation.     

X‐ray Scattering Evaluation of Ultrastructural Changes in Human Dental Tissues with Thermal Treatment

Micro‐ and ultrastructural analysis of burned skeletal remains is crucial for obtaining a reliable estimation of cremation temperature. Earlier studies mainly focused on heat‐induced changes in bone tissue, while this study extends this research to human dental tissues using a novel quantitative analytical approach. Twelve tooth sections were burned at 400–900°C (30‐min exposure, increments of 100°C). Subsequent combined small‐ and wide‐angle X‐ray scattering (SAXS/WAXS) experiments were performed at the Diamond Light Source synchrotron facility, where 28 scattering patterns were collected within each tooth section. In comparison with the control sample, an increase in mean crystal thickness was found in burned dentine (2.8‐fold) and enamel (1.4‐fold), however at a smaller rate than reported earlier for bone tissue (5–10.7‐fold). The results provide a structural reference for traditional X‐ray scattering methods and emphasize the need to investigate bone and dental tissues separately to obtain a reliable estimation of cremation temperature.     

The Stability of 4-Chloromethcathinone in Blood and Vitreous Humor

We present results of our study on the stability of 4-chloromethcathinone (4-CMC) in authentic postmortem peripheral blood and vitreous humor samples. The stability of 4-CMC was determined in postmortem blood samples (for a period of 90 days) and vitreous humor (30 days) at three different temperatures: −15°C, +4°C, and + 23°C. The analyses were carried out using ultra-high-performance liquid chromatography coupled with triple-quadrupole tandem mass spectrometry (UHPLC-QqQ-MS/MS). In both materials, the lowest 4-CMC stability was demonstrated at room temperature. The blood samples stored in a freezer (−15°C) showed stability for the entire study period (90 days), while in the case of the vitreous humor sample stored at the same temperature the concentration of the substance decreased by 53% after 30 days. The study carried out in authentic postmortem blood and vitreous humor samples confirms the previous reports of 4-CMC instability in biological material. Authors suggest that the biological material should be stored frozen until analyses are carried out as soon as possible after collection of the material.     

Scanning Electron Microscopy–Energy‐Dispersive X‐Ray (SEM/EDX): A Rapid Diagnostic Tool to Aid the Identification of Burnt Bone and Contested Cremains

This study investigates the use of Scanning electron microscopy–energy‐dispersive X‐ray (SEM‐EDX) as a diagnostic tool for the determination of the osseous origin of samples subjected to different temperatures. Sheep (Ovis aries) ribs of two experimental groups (fleshed and defleshed) were burned at temperatures of between 100°C and 1100°C in 100°C increments and subsequently analyzed with the SEM‐EDX to determine the atomic percentage of present elements. Three‐factor ANOVA analysis showed that neither the exposure temperature, nor whether the burning occurred with or without soft tissue present had any significant influence on the bone's overall elemental makeup (p > 0.05). The Ca/P ratio remained in the osseous typical range of between 1.6 and 2.58 in all analyzed samples. This demonstrates that even faced with high temperatures, the overall gross elemental content and atomic percentage of elements in bone remain stable, creating a unique “fingerprint” for osseous material, even after exposure to extreme conditions.     

The Effect of Soft Tissue on Temperature Estimation from Burnt Bone Using Fourier Transform Infrared Spectroscopy

This study investigated the effect of soft tissue and different exposure times on the prediction of burning temperatures of bone when using Fourier transform infrared spectroscopy (FTIR). Ovis aries rib bones were burnt at different temperatures and for varying time intervals. Results of a linear regression analysis indicated that burn temperatures can be predicted with a standard error of ±70°C from defleshed bone spectra. Exposure time does not have a significant impact on prediction accuracy. The presence of soft tissue has a significant impact on heat‐induced changes of the bone matrix in low (<300°C) as well as high temperatures (>800°C), slowing down combustion in the former and accelerating it in the latter (p < 0.05). At medium temperatures, no significant difference was noted. These results provide forensic investigators a new perspective with which to interpret the results of crystallinity measures derived from burnt bone.     

Variation in Developmental Time for Geographically Distinct Populations of the Common Green Bottle Fly,Lucilia sericata (Meigen)*

Abstract: Time between death and discovery of remains, or postmortem interval (PMI), can be assessed using blow fly maggot age. Forensic entomologists rely on published, often nonlocal, species‐specific developmental tables to determine maggot age. In a series of common garden experiments, we investigated the developmental rate variation between populations of Lucilia sericata collected from Sacramento, CA, San Diego, CA, and Easton, MA at 16°C, 26°C, and 36°C. For the 16°C trial the time measurement started at egg hatch, while for the higher temperatures the experiment began at oviposition; the wandering stage signified the endpoint for all experiments. The distribution of developmental times differed significantly (ANOVA, p < 0.001) between the three populations within each temperature treatment. We discovered that regional variation of developmental times within a blow fly species exists. This study demonstrates the importance of assembling local population‐specific developmental tables when estimating larval age to determine PMI.     

Comparative Analysis of the Effects of Heat on the PCR‐Amplification of Various Sized DNA Fragments Extracted from Sus Scrofa Molars*

Abstract: This study examined the effects of heat on the amplification of DNA from the dental pulp of Sus scrofa molars and investigated the protection afforded to the pulp tissue by the dental enamel, alveolar process, and soft tissue of the head. Segments of defleshed maxilla and mandible encasing the first molar (n = 60) were subject to a range of temperatures for 15 min. Dental pulps were retrieved. Amplifications using three‐primer and four‐primer multiplexes showed no degradation of the largest fragment following exposure to 450°C. Amplifications in the three‐primer multiplex (283 bp) were successful following exposure to 525°C in maxillary samples only. This study revealed the enamel density of maxillary molars to be greater than mandibular molars in Sus scrofa. Following incineration of intact heads for 15 min (n = 10) and 1 h (n = 4) at an average temperature of 625°C, amplifications of the largest fragment (450 bp) were successful from both maxillary and mandibular teeth.     

Estimating the Temperature of Heat‐exposed Bone via Machine Learning Analysis of SCI Color Values: A Pilot Study

Determining maximum heating temperatures of burnt bones is a long‐standing problem in forensic science and archaeology. In this pilot study, controlled experiments were used to heat 14 fleshed and defleshed pig vertebrae (wet bones) and archaeological human vertebrae (dry bones) to temperatures of 400, 600, 800, and 1000°C. Specular component included (SCI) color values were recorded from the bone surfaces with a Konica‐Minolta cm‐2600d spectrophotometer. These color values were regressed onto heating temperature, using both a traditional linear model and the k‐nearest neighbor (k‐NN) machine‐learning algorithm. Mean absolute errors (MAE) were computed for 1000 rounds of temperature prediction. With the k‐NN approach, the median MAE prediction errors were 41.6°C for the entire sample, and 20.9°C for the subsample of wet bones. These results indicate that spectrophotometric color measurements combined with machine learning methods can be a viable tool for estimating bone heating temperature.     

Color Stability of Dental Restorative Materials Submitted to Heat Sources,for Forensic Purposes

During postmortem examination of the dental arches of carbonized victims, dental restorative materials may be found. The aim of this study was to evaluate the effect of heat source action on the color stability of composite resin (CR) and glass ionomer cement (GIC) restorations, to discriminate between them and compare with antemortem dental data. Sixty bovine teeth (30 CR and 30 GIC) were prepared (6 × 6 × 2 mm) and separated into groups (n = 10). The color readouts were taken by spectrophotometer, before and after heat action (100°C, 200°C, 300°C), in an oven for 15 min. There were color alterations for all coordinates (ΔE, ΔL*, Δa* eΔb*) for both materials. GIC presented greater change. The authors concluded that it is possible to distinguish between the materials by the color changes analyzed by instrumental method, helping victim identification.     

Comparison of new Ampac bags and FireDebrisPAK® bags as packaging for fire debris analysis

Abstract: The FireDebrisPAK^® bags that were produced by Kapak were considered to be one of the best containers for fire debris. Kapak bags were discontinued; however, from July 2010, Ampac is offering a new packaging material. The aim of the presented research was to compare the properties (durability, background interferences, and permeability) of Kapak bags and packaging material offered by Ampac. The analysis was conducted by passive adsorption from the headspace with subsequent thermal desorption and analysis by GC‐MS. The results proved that the properties of the compared materials are similar. Their greatest advantage is that they are impermeable for components of flammable liquids, so there is no danger of losing analytes or cross‐contamination. Their one significant drawback is that they should not be exposed to temperatures above 80°C. At this temperature, they become soft, their inner layer is compromised (becomes sticky), and they emit some volatile organic compounds. Among them, there are compounds that constitute the components of some of flammable liquids.     

The Effect of Temperature and Exposure Time on Stability of Cholesterol and Squalene in Latent Fingermarks Deposited on PVDF Membrane

Cholesterol and squalene are fatty materials of latent fingermarks that can be utilized for dating methodologies and visualization techniques. Previous studies have suggested these compounds undergo degradation in fingermarks as a function of time (days) and light at ambient temperature. However, studies assessing how their composition changes at low and high temperatures over short periods of time (hours) have not been published previously. Here, we performed quantitative analysis of cholesterol and squalene in natural fingermark residue using PVDF membrane, after exposure to a range of temperatures (−20 to 100°C) for 4 and 8 h. We found that levels of both fatty materials remained constant at −20 to 60°C, but both showed significant reduction at 100°C, over short exposure times. These results indicate that cholesterol and squalene are detectable at −20 to 60°C, whereas at 100°C or higher, both are lost due to rapid thermal degradation.     

Quantifying the Temperature of Maggot Masses and its Relationship to Decomposition

Numerous Calliphoridae species have been observed to form larval aggregations during the feeding stage of development, resulting in localized increases in temperature. This study investigates the relationship between maggot numbers in a mass and heat generation. Single‐species aggregations (Lucilia sericata) of various sizes (50–2500 individuals) were reared in the laboratory at a constant ambient temperature of 22°C. Internal mass temperatures were recorded every 5 min throughout the feeding stage of development. Results showed that mass temperatures increased with mass numbers (p‐value < 0.001), ranging from 2.5 to 14°C above ambient. A minimum mass size of 1200 produced overall temperatures that were significantly warmer than ambient, diverging away from 22°C after c. 26 h. These results indicate that the microclimate of a mass has the potential to differ significantly from ambient, which may be influencing larval development rates and should therefore be factored into mPMI estimates to increase accuracy.     

Viability of Human Articular Chondrocytes Harvested Postmortem: Changes with Time and Temperature of In Vitro Culture Conditions

Different studies of long‐term chondrocytes viability have shown a gradual reduction as a function of time and ambient temperature. The aim of our in vitro study was to establish chondrocyte postmortem viability curves for 4°C, 11°C, 23°C, 35°C during 63 days after the donors' death. Osteochondral cylinders were procured from the knees of 16 male donors (20–47 years), stored in preservation media that was not changed, and analyzed in 3‐day intervals using a confocal laser scanning microscope. A significant influence of time on viability was found from Day 9 (p = 0.0029) and onwards (p < 0.0001). The lowest overall chondrocyte viability was at 35°C, followed by 4°C (p < 0.0001). The conditions used in this in vitro analysis suggest that similar viabilities may occur while in situ in the decedent. Further studies of chondrocyte viability from individuals with known postmortem intervals may show premise to help evaluate time since death in the late postmortem interval.     

Pig Organ Energy Loss Comparison Experiments Using BBs

Torso models for ballistics research require that the mechanical properties of simulant materials must match the heterogeneous nature of tissues/organs within the human thorax/abdomen. A series of energy loss experiments were conducted on fresh porcine organs/tissues at room temperature and 37°C, using steel 4.5 mm BBs fired from a Daisy^® brand air rifle. They were compared to FBI and NATO specification ordnance gelatin and a candidate surrogate material called Simulant “A”. Two CED M2 chronographs measured BB velocity. The resulting energy loss was established using KE = 1/2 mv² before and after target perforation. The combined results at room temperature and 37°C were as follows: FBI specification gelatin was similar (p > 0.05) to heart and lung, spleen was similar to NATO specification gelatin, Simulant “A” was similar to hindquarter muscle, and hindquarter muscle, kidney, and spleen were similar to each other regarding energy retardation. These results can be used as a basis for the development of simulant materials to create an anatomically correct heterogeneous model.     

Developmental Times of Chrysomya megacephala (Fabricius) (Diptera: Calliphoridae) at Constant Temperatures and Applications in Forensic Entomology

The characteristic life stages of infesting blowflies (Calliphoridae) such as Chrysomya megacephala (Fabricius) are powerful evidence for estimating the death time of a corpse, but an established reference of developmental times for local blowfly species is required. We determined the developmental rates of C. megacephala from southwest China at seven constant temperatures (16–34°C). Isomegalen and isomorphen diagrams were constructed based on the larval length and time for each developmental event (first ecdysis, second ecdysis, wandering, pupariation, and eclosion), at each temperature. A thermal summation model was constructed by estimating the developmental threshold temperature D₀ and the thermal summation constant K. The thermal summation model indicated that, for complete development from egg hatching to eclosion, D₀ = 9.07 ± 0.54°C and K = 3991.07 ± 187.26 h °C. This reference can increase the accuracy of estimations of postmortem intervals in China by predicting the growth of C. megacephala.