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 Table of Contents  
REVIEW ARTICLE
Year : 2021  |  Volume : 7  |  Issue : 2  |  Page : 54-60

An emergence of dental tissues in the forensic medicine for the postmortem interval estimation: A scoping review


1 Senior Lecturer, Department of Oral Medicine and Radiology, AIMST University, Bedong, Kedah, Malaysia
2 Associate Professor, Department of Maxillo-facial Prosthodontics, Faculty of Dentistry, AIMST Dental Institute, AIMST University, Bedong, Kedah, Malaysia

Date of Submission08-Apr-2020
Date of Decision09-Jul-2020
Date of Acceptance15-Apr-2021
Date of Web Publication26-Jul-2021

Correspondence Address:
Sridevi Ugrappa
Faculty of Dentistry, AIMST Dental Institute, AIMST University, Semeling 08100, Bedong, Kedah
Malaysia
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Source of Support: None, Conflict of Interest: None


DOI: 10.4103/jfsm.jfsm_20_20

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  Abstract 


It is observed that there is not enough literature to support the usefulness of dental tissue in forensic medicine for the estimation of postmortem interval (PMI). Therefore, the authors of this article have systematically reviewed the available literature to understand the most used dental tissues, the methods employed to evaluate these tissues, and their reliability in PMI estimation. A MEDLINE–PubMed search engine was used to review the literature using the keywords “teeth,” “dental tissues,” “post-mortem interval,” and “forensic dentistry” from 2001 until 2020. The most used tissue was dental pulp, but other cellular- and molecular-level dental tissues such as enamel, dentin, cementum, periodontal ligament, odontoblast, and mitochondrial and nuclear DNA and RNA were also used. Based on this observation, the authors have provided the categorical classification to evaluate these dental tissues into morphological, histopathological, and molecular-level examination to understand the reliability of these dental tissues. The molecular examinations to evaluate these tissues were widely employed and emerging in the estimation of PMI as this method could provide a reliable outcome when compared with others. Since the literature reviewed was not enough to answer our questions, however, to conclude, the dental pulp is the widely used tissue so far. Further, more studies are required to authenticate the role of other dental tissues to predict the PMI at an early stage with cost-effective and most convenient techniques.

Keywords: Dental tissues, forensic medicine, forensic odontologist, postmortem interval, time since death


How to cite this article:
Ugrappa S, Jain A. An emergence of dental tissues in the forensic medicine for the postmortem interval estimation: A scoping review. J Forensic Sci Med 2021;7:54-60

How to cite this URL:
Ugrappa S, Jain A. An emergence of dental tissues in the forensic medicine for the postmortem interval estimation: A scoping review. J Forensic Sci Med [serial online] 2021 [cited 2022 May 16];7:54-60. Available from: https://www.jfsmonline.com/text.asp?2021/7/2/54/322338




  Introduction Top


Forensic odontology is deemed as one of the prominent and emerging branches in forensic sciences for decades. It is the branch of forensic medicine which in the interest of justice deals with the proper handling and examination of dental evidence and with proper evaluation and presentation of the dental findings.[1],[2] This specialization plays an important role in the identification of a person by bite mark analysis,[3] DNA/RNA analysis,[4] and sex determination by odontometric measures, rugoscopy, cheiloscopy,[5] etc., The person identification in mass disaster victims such as terrorist attacks and earthquakes and from decomposed, charred bodies such as drowned and deep buried corpses[6] can also be determined from the dental tissues including the estimation of postmortem interval (PMI).

The PMI aka time since death (TSD) is an important element of the forensic investigation and is a critical step in most homicide and unwitnessed death (including hospital deaths). It is the interval between death and time of examination of the body or corpse discovery.[7],[8] After death, many changes begin to take place in the human body due to physicochemical and biochemical processes.[9] These changes progress in an orderly manner until the body disintegrates. Autolytic enzymes initiate the irreversible process of decomposition in the corpse, and simultaneously, putrefying bacteria proceeds with degenerative changes in the body like adipocere and mummification, depending upon the environmental conditions, which can be evaluated by morphological and histopathological changes.[10],[11] The importance of body tissues changes in the deceased person, such as eyes (fall in intraocular pressure, flaccid eyeball, etc.), skin (pallor mortis), muscles (rigor mortis), and liver (livor mortis) have been studied.[12],[13] The measurement of these changes in the deceased along with the time is used to estimate the PMI in medicolegal cases.

The dental tissues have been evaluated in the estimation of PMI. A study was conducted to explain the differences in morphological age-related changes in the teeth in terms of color, translucency, attrition, cemental apposition, secondary dentin, etc., by a computer-assisted image analysis. It was observed that these morphological changes were lower in freshly extracted teeth when compared with the teeth from human skeletal remains, and they concluded that the PMI affects the age-related morphological changes in the teeth.[14] These changes must be taken into consideration while estimating the age of the deceased.

Many factors influence the changes in the tooth after death. Enamel being the hardest tissue in the human body, the morphological changes such as abrasions, cracking as shell appearance, erosions, and mottling are seen which are caused by decomposers. Leaching of environmental factors such as soil and water into the dentinal tissues causes discoloration and crackling of the tissues as well.[15] Soft tissue changes in the labial mucosa and gingival tissues were also studied.[16] Pink tooth phenomenon[17] is also noted because of the leaching of the pulpal hemorrhage into the dentin due to an increase in intracranial blood pressure and imbibition of hemoglobin and its degradation products into the dentinal tubules.

These changes, along with other molecular changes in the dental tissues, may contribute to the estimation of PMI. The present scoping review provides an insight to understand the most used dental tissue, the methods employed to extract these tissues, and their reliability in PMI estimation.


  Methodology Top


A MEDLINE–PubMed search engine was used to review the literature using the keywords “tooth” OR “teeth” AND, “postmortem” AND “interval” AND “forensic dentistry” OR “forensic” AND “dentistry” OR “forensic dentistry” OR “dental” AND “tissues” OR “tissues” from 2001 to 2020. The inclusion criteria comprised the search strategy limited to case reports, comparative study, clinical study, and clinical trial. The articles are published in a language other than English, commentaries, and letters to the editor, and irrelevant articles or articles with inadequate information not included in the review. The study design of the methodology is shown in [Figure 1]. A total of 47,240 articles were displayed during the search, but only 89 articles were collected as these were related to our study, out of which 18 articles were evaluated and reviewed, which met the inclusion criterion of the study.
Figure 1: Flowchart of the review

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  Results Top


It has been observed that the dental tissues are an emerging aid in the estimation of PMI. The most used tissue is found to be dental pulp, but other cellular- and molecular-level dental tissues such as enamel, dentin, cementum, periodontal ligament, odontoblast, mitochondrial DNA (mtDNA) and RNA, and nuclear DNA and RNA have been used in various studies in the estimation of PMI. Based on this observation, the authors have provided the categorical classification to evaluate these dental tissues into morphological, histopathological, and molecular-level examination to understand the reliability of these dental tissues. The molecular examinations to evaluate these tissues are widely employed and emerging in the estimation of PMI [Table 1].
Table 1: Categorical classification of various methods employed to evaluate dental tissues for the estimation of postmortem interval

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  Discussion Top


The determination of the PMI is one of the most challenging problems faced by forensic practitioners. The categorical classification is given for the various methods employed to evaluate the dental tissues for the estimation of PMI. After the literature search, it has been noticed that various dental tissues and methods are used to determine the PMI, which are subsequently discussed [Figure 2].
Figure 2: (a) Morphological changes seen in the decomposed tooth as deposition and cracking of the tooth surface due to environmental factors such as soil decomposers and diatoms, (b) schematic representation of the tooth pulp, for evaluation of morphological changes. (c) Ground section of the tooth for changes in enamel, dentin, and cementum to view histopathological changes, (d) molecular (DNA) contribution of the tooth for postmortem interval estimation

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Morphological evaluation in the tooth structure

Typical morphological features of the tooth are useful in age estimation and person identification, and few studies are conducted to explain the changes in these morphological features as an aid in PMI estimation.

The significance of postmortem tooth loss in PMI estimation was suggested.[18] An animal study was conducted to quantify the decomposition of the periodontal ligament in the exfoliation of incisor teeth using accumulated degree days (ADD) techniques with the help of thermochrons as an estimator of the PMI.[19] The daily average temperature was recorded on an hourly basis using thermochrons for a duration of 6 months. Six teeth were exfoliated during 6 months period, and the average ADD was calculated. This technique could be a potential aid in establishing the length of PMI. However, the reliability of this technique is questionable as contributing factors such as root morphology, oral environment, and malaligned teeth may give false-negative results.

The role of glycoproteins on the surface of the tooth and other body remains could be an indicator for the estimation of PMI. The glycoprotein adhesions deposits by Balanus improvisus on the human skeletal and dental remains were in the fluvial environment for up to the range of 375–410 days. Further studies should be carried out on their predictability of life cycle and decay rate of adhesions to consider as a tool for PMI estimation.[20]

The importance of colorimetric analysis in the morphological changes in the pulp has been mentioned. The morphological changes in the color of pulp were observed, and the quantitative measure was done up to 84 days using a simple colorimetric assay. It was concluded that this assay can be used to estimate the PMI.[21]

Mehendiratta et al.[22] and Bhuyan et al.[10] have observed the gradual morphological changes in the dental pulp as soft to firm at the initial stage, followed by jelly-like structure, semi-viscid fluid and friable at the later stages, till the pulp ceases to exist. The coastal environment has an influence on the dental pulp for these changes to occur and can be appreciated up to 6 days as observed by Mehendiratta et al. However, the similar changes in the dental pulp can be appreciated by up to 2 years if studied in normal environmental conditions as studied by Bhuyan et al.[10]

Diatoms on the surface of the tooth and human remains may be a valuable tool in the estimation of PMI. A study was conducted to evaluate the diatoms attached to the surface of dental enamel to determine PMI for underwater corpses by the Electron Probe X-ray Micro Analyzer.[23] It was observed that as the immersion time increased, the quantity of seawater constituents such as O, Si, Mg, K, Al, and S detected on the surface of dental enamel increased, while the quantity of the main dental components (Ca and P) that were detected gradually decreased. This method was unique for estimating the PMI as other conventional methods depending on the degree of decomposition of the corpse. The authors of this study claimed that this method is a breakthrough technique for evaluating the TSD more objectively, compared to the conventional method of determination based on the degree of decomposition of the corpse; therefore, the concurrent use of this method may increase the accuracy of PMI.

An animal study was conducted to explain the importance of morphological changes such as the alteration in the mineral density and the surface abrasion of the tooth in the estimation of PMI. The micro-computed tomographic scan analysis results showed that there were decrease in the mineral density of the enamel tissues and increase in surface abrasion of the hard tissues, but the thickness of the enamel and restorative cement remained the same.[24] Thus, the evaluation of these parameters can be considered in PMI determination.

The changes in mechanical properties of the restorative materials in a marine environment were evaluated to observe the significant changes in the microhardness and surface roughness of restorative materials, and the results showed that there was a decrease in microhardness and increase in surface roughness of restorative material, and these findings could be helpful in the PMI estimation.[25]

Histopathological evaluation in the tooth structure

Several studies have evaluated the dental tissue in the estimation of the PMI by employing histopathological methods. A study was conducted to explain the microscopic changes of odontoblastic cells in PMI estimation.[26] The odontoblastic cells are fixed postmitotic cells and are the most characteristic of the dentin–pulp complex. The tooth was extracted during the autopsy procedure every day and stained with hematoxylin and eosin after decalcification. Densities of odontoblasts in the dental pulp were counted using light microscopy. They have observed that the reduced number of odontoblasts and histological appearance of these cells may be one of the additional parameters in the estimation of PMI in the early postmortem period for up to 5 days. Since the sample size of the study was less, it could not be a reliable tool in the estimation of PMI.

The histopathological features in the dental pulp were seen as nuclear degenerative changes and a smaller number of fibroblasts at the end of 48 h and process of karyolysis at 72 h. At 120 h, pulp showed predominantly vacuolated and no evidence of any nuclear debris. Pulp completely putrefied at the end of 144 h.[22] Various studies[27],[28] have been conducted to determine the effect of environmental factors such as average daily rainfall precipitation, temperature, soil humidity, soil temperature, and soil pH, on the rate of putrefaction of the dental pulp and hence has to be considered during PMI. Few studies were also evaluated the histopathological postmortem changes in the labial mucosa[29] and human gingival tissues[16] to correlate these changes with PMI estimation. The loss of structural changes was noticed from epithelium to cytoplasmic to nuclear changes as the period of the PMI increased. With this, they concluded that decomposition at the cellular level started within 24 h of death and other features will start decomposing subsequently and gingival tissues can be used in PMI estimation in the early postmortem period. However, the climatic and seasonal variations must be considered while taking these tissues for PMI estimation, as these tissues get fibrosed at a later stage after 24 h. Another study was conducted to estimate the PMI from antimortem gingival tissue in histological changes over a time interval of 15 min to 4 h and to correlate these histological findings at an early stage of duration. There was a positive correlation between the time interval and the appearance of the histological changes.[30] However, certain environmental factors and size of samples were not considered in this study, which also plays a crucial role in the autolysis of the tissues.

One study has evaluated the histological examination of dental tissues for the estimation of PMI.[31] The dentin and pulp were destroyed in a relatively short period. The structural changes in the predentin layer were lost at the period of 16 months, but these changes were seen in the cementum at the same time. It was concluded that the structural changes of cementum were less affected than dentin in the buried teeth.

Later, a study was conducted to explain the qualitative and quantitative method of retrieval of dental pulp tissue for the estimation of PMI. This new methodology has overcome the limitation of the previous studies in which changes in the dental pulp were seen up to 6 days. The histological analysis was performed over a period interval of 24 h, 1 month, 3 months, and 6 months. This study has explained the postmortem changes such as a decrease in the percentage of a number of nuclei and an increase in the collagen fibers as PMI increases up to 6 months as a PMI indicator.[32]

In 2020, an observational study of microbiological characterization in tooth pulp was done by Bhuyan et al.,[10] and they mentioned that microbiological assessment for staphylococci and streptococci by Gram staining was seen up to 2 years. A study by Hyde et al.[33] noticed a change in aerobic species to anaerobic flora in the prebloat mouth swab. However, there is a lack of conclusive results in the literature, and furthermore, studies are required.

Molecular evaluation in the tooth structure

During postmortem autolysis, cellular organelles and nuclear DNA break down into their constituent parts. The importance of DNA degradation of pulp tissue was investigated in PMI estimation by using flow cytometry. The pulp was removed during the interval from 24 to 144 h postextraction. In this study, pulp tissue was found to be unreliable to determine the early PMI but might be of greater value in the later stages.[34]

Another study explained the correlation between DNA degradation of costicartilage and dental pulp cells with the PMI by using image cytometry from 0 to 15 days. The DNA content of both the tissues was degraded with the prolongation of the PMI.[35]

A study was conducted to assess the quantity, quality, and stability of the dental DNA using real-time polymerase chain reaction (RT-PCR), and it was observed that the stability of the dental DNA decreased over time, differently affecting the quality and quantity of DNA in a time-dependent process over the first 18 months postextraction, and concluded that these results have potential use in PMI estimation.[36] Conversely, Raimann et al. concluded that there is no significant difference in the amount of DNA obtained over a period of time duration.[37]

Later, in 2013, an animal study was conducted on the tooth pulp of pigs to examine the time-dependent differences in RNA decay rate to extend the time frame over which early PMI estimates can be made.[21] Comparing the decay rates of a large, labile segment of β-actin RNA and a smaller, more stable, nonoverlapping segment of the same RNA from tooth pulp, the researchers were able to estimate PMI values of pigs buried within a shallow grave for up to 84 days.

Different tissues have differential survival rates of nuclear and mitochondrial DNA (mtDNA). The advanced genetic analysis to examine the DNA content and rate of DNA degradation from the human molar teeth for estimation of PMI was performed.[31] The DNA was extracted from coronal dentin, root dentin, cementum, and pulp of 114 teeth. The RT quantification assays showed nuclear DNA and mtDNA degraded exponentially with different rates, depending on PMI and soil temperature. It was confirmed by the short tandem repeats-based genotyping method that cementum from teeth buried up to 16 months can provide a reliable amount of source of nuclear DNA, without the need for specialized equipment or large-volume demineralization steps.

The confined nature of dental pulp tissue makes it an ideal candidate for PMI estimation, as the impact of environmental factors is reduced. Using this concept, a study was conducted to analyze the rate of RNA degradation in human dental pulp.[38] The RNA integrity analysis determined the time of PMI with a high confidence level in the first 21 days. Apart from this, the crude estimation of incubation time of teeth between 20 and 42 days was also performed with the help of PCR-based technique. These results showed that this method might be a promising new tool for PMI estimation despite the limitations like of inclusion of healthy teeth and did not assess the effect of environmental factors.

The dental DNA-based identification has a crucial effect on the elapsed PMI estimation. A study was conducted to evaluate the effect of antemortem and postmortem factors on dental DNA-based.[39] The amount of DNA concentration was evaluated in six different postmortem conditions using RT-PCR. It was found that yield of DNA was best in the first 10 days, and later, it decreased dramatically in the following time period. Further, it was noted that teeth extracted from burnt and fresh corpses yielded the highest amount of DNA, while skeletonized exhumed corpses resulted in the lowest DNA amount. Indeed, dry and indoor conditions demonstrated better results than those in water, outdoors, or buried in the ground. On the other hand, antemortem factors including sex, age, tooth type, and tooth root portions did not reveal a significant effect on dental DNA yield. It was suggested that antemortem factors are considerably more subjected to individual variation while postmortem factors including PMI, postmortem conditions, and the relevant surrounding environments have a considerable effect on the dental DNA amount yielded.

Another study was conducted to evaluate the DNA concentration using a field-forward rapid DNA identification system.[40] It was observed that using this technology DNA concentration can be used from human remains, tooth, and buccal tissue in 2 h, 12 months, and 3–11 days, respectively, for PMI estimation.


  Conclusion Top


The PMI plays a major role in forensic science for the identification of a suspect. Currently, a few studies have been conducted to demonstrate the importance of dental tissues in the estimation of PMI, in which most of the studies are related to dental pulpal tissue and molecular structure of tooth DNA by PCR method was used. However, still, there is a deficit in satisfactory conclusive results to state the standardized technique for using a particular dental tissue in PMI estimation. Further, more studies are required to authenticate the role of other dental tissues to predict the PMI at an early stage with cost-effective and most convenient techniques.

Financial support and sponsorship

Nil.

Conflicts of interest

There are no conflicts of interest.



 
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