Estimation of vitality of skin injuries by immunohistochemistry using TNF-α
https://doi.org/10.31549/2542-1174-2026-10-1-21-31
Abstract
I n t r o d u c t i o n . Estimation of wound vitality is one of the most challenging and urgent tasks in forensic medicine, having undeniable significance. Conventional histological methods, based on the assessment of inflammatory response elements, are often insufficiently sensitive for diagnosing injuries inflicted shortly before death. The development and implementation of highly sensitive molecular morphological methods is an important direction for advancing expert practice.
A i m . To evaluate the diagnostic value of such immunohistochemical marker as tumor necrosis factor-alpha (TNF-α) and the method of quantitative digital analysis for the objective determination of the vitality of skin blunt force injuries.
M a t e r i a l s a n d m e t h o d s . Skin samples from 14 cadavers with documented antemortem injuries (28 sections, 140 fields of view) were analyzed. Samples were taken from the area of external injury, and a control tissue sample was also taken from an undamaged area. Paraffin-embedded sections were stained with monoclonal antibodies to TNF-α. Digital images were analyzed using Fiji software using color deconvolution to isolate DAB staining and subsequently determine the area of marker expression in the epidermis and dermis separately.
R e s u l t s . A statistically significant increase in the area of TNF-α expression was found in the damaged area compared to intact skin in control samples. In the epidermis, the difference in medians was –16,66% (p = 0,0002), and in the dermis –7,93% (p = 0,0072).
C o n c l u s i o n . Immunohistochemical detection of TNF-α in combination with quantitative analysis of digital image is highly informative and objective method for ascertaining the skin wound vitality. This approach allows for the identification of early molecular signs of alteration preceding morphological changes and minimizes the subjective factor during the interpretation of results, which increases the evidentiary value of the forensic medical report.
About the Authors
V. N. Metr
Gomel State Medical University
Belarus
Belarus
Vitaliy N. Metr – Senior Lecturer, Department of Pathological Anatomy
5, Lange str., Gomel, 246000
E. M. Kildyushov
Pirogov Russian National Research Medical University
Russian Federation
Russian Federation
Evgeniy M. Kildyushov – Dr. Sci. (Med.), Professor, Head, Department of Forensic Medicine named after P.A. Minakov, Institute of Human Biology and Pathology
Moscow
S. N. Likhacheva
Department of Forensic Histological Examinations, Directorate General of Forensic Medical Examinations, Central Office of the State Forensic Examination Committee of the Republic of Belarus
Belarus
Belarus
Svetlana N. Likhacheva – Head
Minsk
A. Yu. Krylov
Institute for Advanced Training and Retraining of Personnel, State Forensic Examination Committee of the Republic of Belarus
Belarus
Belarus
Andrey Yu. Krylov – Cand. Sci. (Med.), Associate Professor, Head, Department of Forensic Medicine, Institute for Advanced Training and Retraining of Personnel
Minsk
References
1. Kupriyanov D.D., Fedulova M.V. Detection of some markers of the early phase of the inflammatory and reparative process in mechanical skin injuries. Forensic Medical Expertise. 2024;67(2):17-19. DOI: 10.17116/sudmed20246702117. (In Russ.)
2. Pigolkin Yu.I., Kupriyanov D.D., Kopylov A.V. et al.; ed. by Yu.I. Pigolkin (2018). Forensic Medicine: National Guidelines. Мoscow: GEOTAR-Media, 2018. 576 p. (In Russ.)
3. Surkova E.I. Wound age estimation by immunohistochemical method. Medical Examination Problems. 2014;14(4(56)):12-14. (In Russ.)
4. Fedulova M.V., Kupriyanov D.D. Reliability of the immunohistochemical estimation of vitality and injury age: analysis and perspectives for study. Forensic Medical Expertise. 2020;63(2):52-57. DOI: 10.17116/sudmed20206302152. (In Russ.)
5. Ohshima T. Forensic wound examination. Forensic Sci. Int. 2000;113(1-3):153-164. DOI: 10.1016/s0379-0738(00)00269-3.
6. Cecchi R. Estimating wound age: looking into the future. Int. J. Legal Med. 2010;124(6):523-536. DOI: 10.1007/s00414-010-0505-x.
7. Grellner W., Madea B. Demands on scientific studies: vitality of wounds and wound age estimation. Forensic Sci. Int. 2007;165(2-3):150-154. DOI: 10.1016/j.forsciint.2006.05.029.
8. Wenzlow N., Mills D., Byrd J. et al. Review of the current and potential use of biological and molecular methods for the estimation of the postmortem interval in animals and humans. J. Vet. Diagn. Invest. 2023;35(2):97-108. DOI: 10.1177/10406387231153930.
9. Crowe A.R., Yue W. Semi-quantitative determination of protein expression using immunohistochemistry staining and analysis: an integrated protocol. Bio Protocol. 2019;9(24):e3465. DOI: 10.21769/BioProtoc.3465.
10. Idriss H.T., Naismith J.H. TNF alpha and the TNF receptor superfamily: structure-function relationship(s). Microsc. Res. Tech. 2000;50(3):184-195. DOI: 10.1002/1097-0029(20000801)50:3<184::AIDJEMT2>3.0.CO;2-H.
11. Panwar R., Shekhawat R.S., Shukla K.K. et al. Quantitative estimation of TNF-α and IL-3 by using ELISA from human lung tissue in fatal asphyxial deaths. J. Forensic Leg. Med. 2023;98:102559. DOI: 10.1016/j.jflm.2023.102559.
12. Spiekstra S.W., Toebak M.J., Sampat-Sardjoepersad S. et al. Induction of cytokine (interleukin-1alpha and tumor necrosis factor-alpha) and chemokine (CCL20, CCL27, and CXCL8) alarm signals after allergen and irritant exposure. Exp. Dermatol. 2005;14(2):109-116. DOI: 10.1111/j.0906-6705.2005.00226.x.
13. Zinovkin D.A. The role of tumor inflammatory component of the tumor microenvironment of the endometrioid adenocarcinoma of the corpus uteri in different disease outcomes. Tumors of Female Reproductive System. 2016;12(1):73-78. DOI: 10.17650/1994-4098-2016-12-1-73-78. (In Russ.)
14. Fiji. URL: https://imagej.net/software/fiji/#publication (accessed 14.12.2025).
Review
For citations:
Metr V.N., Kildyushov E.M., Likhacheva S.N., Krylov A.Yu. Estimation of vitality of skin injuries by immunohistochemistry using TNF-α. Journal of Siberian Medical Sciences. 2026;(1):21-31. (In Russ.) https://doi.org/10.31549/2542-1174-2026-10-1-21-31
Views:
30
JATS XML
Email this article 