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An Official Publication of the Indian Association of Oral and Maxillofacial Pathologists

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Year : 2020  |  Volume : 24  |  Issue : 3  |  Page : 568-571

Estimation of postmortem death interval from autopsied tongue tissue: A cross-sectional study

1 Department of Oral Pathology and Microbiology, Sree Balaji Dental College and Hospital, Bharath Institute of Higher Education and Research, Chennai, Tamil Nadu, India
2 Department of Conservative and Endodontics, Sree Balaji Dental College and Hospital, Chennai, Tamil Nadu, India
3 Department of Forensic Medicine, Government Thiruvarur Medical College, Thiruvarur, Tamil Nadu, India
4 Department of Oral Pathology and Microbiology, Sathyabama Dental College and Hospital, Chennai, Tamil Nadu, India
5 Department of Oral Pathology and Microbiology, V. S. Dental College and Hospital, Bengaluru, Karnataka, India

Date of Submission24-Nov-2020
Date of Acceptance25-Nov-2020
Date of Web Publication09-Jan-2021

Correspondence Address:
S Rajkumari
Research Scholar, Bharath Institute of Higher Education and Rresearch, Chennai, Tamilnadu
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Source of Support: None, Conflict of Interest: None

DOI: 10.4103/jomfp.jomfp_479_20

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Context: Estimation of time since death is the preliminary step in any postmortem examination. Although there are various physiological methods to conclude the postmortem, interval histological changes can be applied to obtain precision. However, the utility of oral tissues for such an event is still evolving.
Aims: The present study was conducted to assess the efficacy of postmortem histological changes that occur in tongue to conclude the postmortem interval (PMI).
Materials and Methods: After obtaining institutional human ethical committee, tongue tissue was collected for during routine autopsy procedure. The study comprised twelve autopsied tongue tissues. The tissue specimens were subjected to routine laboratory tissue processing procedure and the histological changes were evaluated.
Conclusion: This is the first study of this kind in the scientific literature to explore the tongue tissue to estimate the PMI. There were definite changes in the epithelium and the connective tissue of the tongue, and these features were highly remarkable at various postmortem time intervals.

Keywords: Death interval, histology, oral autopsy, postmortem, time since death, tongue

How to cite this article:
Rajkumari S, Mensudar R, Naveen N, Thayumanavan B, Thammaiah S. Estimation of postmortem death interval from autopsied tongue tissue: A cross-sectional study. J Oral Maxillofac Pathol 2020;24:568-71

How to cite this URL:
Rajkumari S, Mensudar R, Naveen N, Thayumanavan B, Thammaiah S. Estimation of postmortem death interval from autopsied tongue tissue: A cross-sectional study. J Oral Maxillofac Pathol [serial online] 2020 [cited 2021 Jan 21];24:568-71. Available from: https://www.jomfp.in/text.asp?2020/24/3/568/306665

   Introduction Top

Forensic odontology usually involves personal identification and crime investigation through comparative and reconstructive dental identification mainly by analyzing antemortem and postmortem dental records, dental profiling, bitemark, cheiloscopy, rugoscopy, dental DNA analysis, facial reconstruction and superimposition. Most of these procedures involve hard tissue component of the facial and dental apparatus, while the soft tissue analysis is rarely sought (rugoscopy). Histological analysis of the oral hard tissues is proved and well established in age estimation;[1] however, microscopic analysis of oral soft tissues in at postmortem examination is at its primitive stage. The most important medicolegal issue in any postmortem examination is to determine time elapsed since death which is also referred as postmortem interval(PMI),[2],[3] and still there are many researches in this regard to get appropriate time of death.[4] Although the physical signs such as algor mortis, rigor mortis, livor mortis, muscle excitability, gastric content, retinal, iris changes and biochemical changes can elucidate the time since death,[4],[5] microscopic changes can be considered as an appropriate method.[6],[7],[8] Hence, in this study, we made an attempt to observe and correlate the various changes in the histology of the tongue with the PMI.

   Materials and Methods Top

The study was conducted after obtaining human ethical clearance from Chengalpattu Medical College and Hospital. The study sample included twelve tongue tissue samples collected during routine autopsy procedure at the mortuary in Chengalpattu Government Hospital, Tamil Nadu. The relatives and the investigating police officer were informed about the study details, and written consent was obtained from them. Demographic data and postmortem number of the corpse were collected from the relatives, and there was no gender discrimination. Inclusion criteria considered for the study are as follows: corpse in which death occurred due to road traffic accident without injury to the face and jaws specifically related to tongue. Exclusion criteria comprised all other cases reported to the department of forensic medicine for autopsy, including road traffic accident with injury to the face and jaws related to the study site, pathological lesion involving the study site and frozen corpse. All the twelve specimens were collected at varied time intervals of death.

The specimens were immediately washed under running water and transferred to a container with 10% formalin for fixation. Routine tissue processing was done and stained with hematoxylin and eosin. The microscopic examination of the tissue changes was carried out by two individual observers, and they were blinded to avoid any bias. The histopathological changes at architectural and cellular level were tabulated and correlated with the time interval of death as reported by Yadhav A et al., 2012.[9]

   Results Top

The observed histological changes in the epithelium and the connective tissue of the tongue are given in [Table 1]. The histological changes could be categorized as architectural changes, nuclear changes and cytoplasmic changes. The architectural changes include epithelial disruption, shredding and separation of epithelium from the connective tissue and are shown in [Figure 1]. Nuclear changes and cytoplasmic changes include pyknosis, vacuolation, karyolysis, karyorrhexis, cytoplasmic vacuolation, cytoplasmic eosinophilia and homogenization and are given in [Figure 2] and [Figure 3]. Early stages (0–8 hrs) of PMI show minimal changes at the cellular level; the nuclear changes and the cellular changes are more evident at 8–16 hrs; while cytoplasmic and nuclear alterations, architectural changes are exceedingly conspicuous at 16–24 hrs. The interobserver variability was calculated for various histological changes and is given in [Table 2].
Table 1: Histopathological changes in tongue at different postmortem interval

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Figure 1: Hematoxylin and eosin-stained tongue tissue section (16–24 hrs) at low-power view (×10) shows (a) epithelial shredding and disruption, (b) split between the epithelium and connective tissue, (c) cytoplasmic eosinophilia, homogenization and karyolysis in the superficial layer and (d) homogenization and clumping of collagen in connective tissue

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Figure 2: Hematoxylin and eosin-stained tongue tissue section (16–24 hrs) at high-power view (×40) shows (a) karyolysis, (b) pyknosis and (c) nuclear vacuolation

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Figure 3: Hematoxylin and eosin-stained tongue tissue section (8–16 hrs) at high-power view (×40) shows (a) cytoplasmic vacuolation and (b) ballooning of epithelial cells at the spinous layer

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Table 2: Kappa test for inter-observer variability

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   Discussion Top

Death of a person under suspicious condition demands enormous queries about the nature of the incident such as the cause of death and time of death. Although careful physiological examination can unveil the mystery, the forensic experts at time seek specialist opinion to demystify some unknown fact.

Once the vital functions are halted, hypoxia is preceded by ischemia, leading to downregulation of oxidative respiration, which at end stage causes irreversible cell injury and cell death. The cell death occurs due to mitochondrial dysfunction, followed by rupture of lysosomal membranes, resulting in enzymatic action.[4] These changes are sequential and may be correlated with PMI.

The microscopic changes of tongue tissue observed in our study are consistent with the study by Yadhav et al.,[9],[10] They have reported cytoplasmic eosinophilia, homogenization and mild nuclear changes in the superficial layers of the epithelium at early stages of post-mortem (before 8 hrs). Beyond 8hrs nuclear changes such as pyknosis, vacuolation, karyolysis, karyorrhexis and cytoplasmic vacuolation were evident in the spinous layer and mild nuclear changes in the superficial layers of the epithelium at early stages of postmortem, i.e., before 8 h, whereas nuclear changes such as pyknosis, vacuolation, karyolysis, karyorrhexis and cytoplasmic vacuolation were evident in the spinous layer as the PMI prolongs (8–16 h). Epithelial disruption, shredding, separation of epithelium from the connective tissue and clumping of collagen fiber are more evident in the later stages of PMI (16–24 hrs). In addition to these findings, we noticed degradation of muscle tissue in the deeper layers beyond 16 h. In the post-mortem study on gingival tissue by Pradeep et al.,[11] cytoplasmic eosinophilia and homogenization that was noticed within 10 hrs, faded as the time elapsed similar to our study. Whereas cytoplasmic vacuolation present at all stages was more appreciated beyond 20 hrs. They observed nuclear changes at all time intervals in the spinous layer, but intensity varied as the time elapsed. They considered PMI interval of 10 hrs, which contradicted our study (8 hrs time interval), but the findings in both were parallel. Mahalakshmi et al., 2016, in their study on antemortem gingival tissue fixed the tissue at the time range of 15 min, 30 min, 45 min, 1 hrs, 2 hrs and 4 hrs; however, substantial microscopic changes such as homogenization loss of epithelial architecture and nuclear changes were evident at 4th hrs.[12] Muthukrishnan et al. in their study on postmortem gingival specimen considered the time frame of 2 hrs and 4 hrs. In their study, they observed cytoplasmic homogenization and other cytoplasmic and nuclear changes within 2 hrs, but there was no significant difference at 4th hr.[13]

The interobserver variability test shows that there was 100% (k = 1.000) agreement between the observers with regard to nuclear and connective tissue changes, while 80% (k = 0.824) agreement in case of architectural alterations and cytoplasmic changes brought moderate agreement with k value of 0.556, which shows that there is slight confusion over these. The dispute in the interobserver error can be reduced by involving only one researcher for selected variables, restricting the study to the researcher only and repeated review of the parameter.[14]

The tongue tissue comprising numerous papillae is a specialized mucosa, which shows varied degeneration pattern similar to other types of oral mucosa and can be taken as reliable marker of PMI estimation.

   Conclusion Top

Applicability of microscopic alterations in PMI determination is still at its primitive, and this is the first study in the scientific literature to explore the microscopy of tongue tissue for such an event. This study gives a path to future forensic pathologists to consider microscopic changes in tongue as a factor to determine PMI.

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Conflicts of interest

There are no conflicts of interest.

   References Top

Shafer WG, Hine MK, Levy BM. Forensic Odontology. In: Sivapathasundharam B, editor. Shafer's Textbook of Oral Pathology. 8th ed. India: Elsevier; 2016. p. 716-40.  Back to cited text no. 1
Abdelaleem SA, Younis RH, Ahmed RF. Postmortem interval estimation using myoglobin concentration in different glandular tissues. Int J Forensic Sci Pathol 2016;4:259-65.  Back to cited text no. 2
Brooks JW. Postmortem changes in animal carcasses and estimation of the postmortem interval. Vet Pathol 2016;53:929-40.  Back to cited text no. 3
Vavpotic M, Turk T, Martincicc DS, Balazic J. Characteristics of the number of odontoblasts in human dental pulp post-mortem. Forensic Sci Int 2009;193:122-6.  Back to cited text no. 4
Rutty GN, Essentials of Autopsy Practice – Current Methods and Modern Trends. London: Springer; 2006.  Back to cited text no. 5
Dettmeyer RB. The role of histopathology in forensic practice: An overview. Forensic Sci Med Pathol 2014;10:401-12.  Back to cited text no. 6
Erlandsson M, Munro R. Estimation of the post-mortem interval in beagle dogs. Sci Justice 2007;47:150-4.  Back to cited text no. 7
Bryant BH, Boekelheide K. Time-dependent chanjonges in post-mortem testis histopathology in the rat. Toxicol Pathol 2007;35:665-71.  Back to cited text no. 8
Yadav A, Angadi PV, Hallikerimath S, Kale A, Shetty A. Applicability of histologic post-mortem changes of labial mucosa in estimation of time of death – A preliminary study. Aust J Forensic Sci 2012;44:343-52.  Back to cited text no. 9
Yadav AB, Angadi PV, Kale AD, Yadav SK. Histological assessment of cellular changes in postmortem gingival specimens for estimation of time since death. J Forensic Odontostomatol 2015;33:19-26.  Back to cited text no. 10
Pradeep GL, Uma K, Sharada P, Prakash N. Histological assessment of cellular changes in gingival epithelium in ante-mortem and post-mortem specimens. J Forensic Dent Sci 2009;1:61-5.  Back to cited text no. 11
  [Full text]  
Mahalakshmi V, Gururaj N, Sathya R, Sabarinath TR, Sivapathasundharam B, Kalaiselvan S. Assessment of histological changes in antemortem gingival tissues fixed at various time intervals: A method of estimation of postmortem interval. J Forensic Dent Sci 2016;8:114.  Back to cited text no. 12
[PUBMED]  [Full text]  
Muthukrishnan S, Narasimhan M, Paranthaman SK, Hari T, Viswanathan P, Rajan ST. Estimation of time since death based on light microscopic, electronmicroscopic, and electrolyte analysis in the gingival tissue. J Forensic Dent Sci 2018;10:34-9.  Back to cited text no. 13
[PUBMED]  [Full text]  
Gordon CC, Bradtmiller B. Interobserver error in a large scale anthropometric survey. Am J Hum Biol 1992;4:253-63.  Back to cited text no. 14


  [Figure 1], [Figure 2], [Figure 3]

  [Table 1], [Table 2]


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