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Immunohistochemical analysis on cyclooxygenase-2 for wound age determination

International Journal of Legal Medicine Aims and scope Submit manuscript

Abstract

Immunohistochemical study combined with morphometry was carried out to examine the expression of cyclooxygenase-2 (COX-2) using 60 human skin wounds of different ages: group I, 0–4 h (n = 11); II, 8 h–2 days (n = 21); III, 3–9 days (n = 14); and IV, 12–21 days (n = 14). In wound specimens aged 2 h to 2 days, anti-myeloperoxidase-positive neutrophils observed at the wound site expressed immunopositive reaction to COX-2. In wound specimens of more than 3 days, CD68-positive macrophages as well as neutrophils were positively immunostained with anti-COX-2. In group II, all 21 wound samples had COX-2-positive ratios of >40 %, and 15 out of them showed >50 %. In group III, only three wound samples with the postinfliction intervals of 3 days showed positive ratios of 40–50 % and the remaining 11 cases less than 40 %. In groups I and IV, all 25 wound specimens had COX-2-positive ratio of <40 %. With regard to the practical applicability with forensic safety, these observations suggested that a COX-2-positive ratio of >40 % indicated a wound age of 8 h to 3 days. Moreover, COX-2-positive ratios, considerably exceeding a ratio of 50 %, indicate a wound age of 8 h to 2 days. Collectively, COX-2 would be a useful marker for the determination of early wound age.

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References

  1. Kondo T (2007) Timing of skin wounds. Leg Med (Tokyo) 9:109–114

    Article  Google Scholar 

  2. Kondo T, Ishida Y (2010) Molecular pathology of wound healing. Forensic Sci Int 203:93–98

    Article  PubMed  CAS  Google Scholar 

  3. Cecchi R (2010) Estimating wound age: looking into the future. Int J Legal Med 124:523–536

    Article  PubMed  Google Scholar 

  4. Betz P (1994) Histological and enzyme histochemical parameters for the age estimation of human skin wounds. Int J Legal Med 107:60–68

    Article  PubMed  CAS  Google Scholar 

  5. An JL, Ishida Y, Kimura A, Kondo T (2011) Immunohistochemical examination of intracerebral aquaporin-4 expression and its application for differential diagnosis between freshwater and saltwater drowning. Int J Legal Med 125:59–65

    Article  PubMed  Google Scholar 

  6. An JL, Ishida Y, Kimura A, Kondo T (2010) Forensic application of intrarenal aquaporin-2 expression for differential diagnosis between freshwater and saltwater drowning. Int J Legal Med 124:99–104

    Article  PubMed  Google Scholar 

  7. Nosaka M, Ishida Y, Kimura A, Kondo T (2010) Immunohistochemical detection of MMP-2 and MMP-9 in a stasis-induced deep vein thrombosis model and its application to thrombus age estimation. Int J Legal Med 124:439–444

    Article  PubMed  Google Scholar 

  8. Nosaka M, Ishida Y, Kimura A, Kondo T (2009) Time-dependent appearance of intrathrombus neutrophils and macrophages in a stasis-induced deep vein thrombosis model and its application to thrombus age determination. Int J Legal Med 123:235–240

    Article  PubMed  Google Scholar 

  9. Fracasso T, Pfeiffer H, Michaud K, Köhler H, Sauerland C, Schmeling A (2011) Immunohistochemical expression of fibronectin and C5b-9 in the myocardium in cases of carbon monoxide poisoning. Int J Legal Med 125:377–384

    Article  PubMed  Google Scholar 

  10. Yoshida C, Ishikawa T, Michiue T, Quan L, Maeda H (2011) Postmortem biochemistry and immunohistochemistry of chromogranin A as a stress marker with special regard to fatal hypothermia and hyperthermia. Int J Legal Med 125:11–20

    Article  PubMed  Google Scholar 

  11. Bohnert M, Anderson J, Rothschild MA, Böhm J (2010) Immunohistochemical expression of fibronectin in the lungs of fire victims proves intravital reaction in fatal burns. Int J Legal Med 124:583–588

    Article  PubMed  Google Scholar 

  12. Dressler J, Hanisch U, Kuhlisch E, Geiger KD (2007) Neuronal and glial apoptosis in human traumatic brain injury. Int J Legal Med 121:365–375

    Article  PubMed  CAS  Google Scholar 

  13. Kondo T, Tanaka J, Ishida Y, Mori R, Takayasu T, Ohshima T (2002) Ubiquitin expression in skin wounds and its application to forensic wound age determination. Int J Legal Med 116:267–272

    Article  PubMed  CAS  Google Scholar 

  14. Ishida Y, Kimura A, Takayasu T, Eisenmenger W, Kondo T (2008) Expression of oxygen-regulated protein 150 (ORP150) in skin wound healing and its application for wound age determination. Int J Legal Med 122:409–414

    Article  PubMed  CAS  Google Scholar 

  15. Ishida Y, Kimura A, Takayasu T, Eisenmenger W, Kondo T (2009) Detection of fibrocytes in human skin wounds and its application for wound age determination. Int J Legal Med 123:299–304

    Article  PubMed  Google Scholar 

  16. Kondo T, Ohshima T, Eisenmenger W (1999) Immunohistochemical and morphometrical study on the temporal expression of interleukin-1α (IL-1α) in human skin wounds for forensic wound age determination. Int J Legal Med 112:249–252

    Article  PubMed  CAS  Google Scholar 

  17. Hayashi T, Ishida Y, Kimura A, Takayasu T, Eisenmenger W, Kondo T (2004) Forensic application of VEGF expression to skin wound age determination. Int J Legal Med 118:320–325

    Article  PubMed  Google Scholar 

  18. Kondo T, Ohshima T, Mori R, Guan DW, Ohshima K, Eisenmenger W (2002) Immunohistochemical detection of chemokines in human skin wounds and its application to wound age determination. Int J Legal Med 116:87–91

    Article  PubMed  CAS  Google Scholar 

  19. Ma WX, Yu TS, Fan YY, Zhang ST, Ren P, Wang SB, Zhao R, Pi JB, Guan DW (2011) Time-dependent expression and distribution of monoacylglycerol lipase during the skin-incised wound healing in mice. Int J Legal Med 125:549–558

    Article  PubMed  Google Scholar 

  20. Yu TS, Cheng ZH, Li LQ, Zhao R, Fan YY, Du Y, Ma WX, Guan DW (2010) The cannabinoid receptor type 2 is time-dependently expressed during skeletal muscle wound healing in rats. Int J Legal Med 124:397–404

    Article  PubMed  Google Scholar 

  21. Betz P (1995) Immunohistochemical parameters for the age estimation of human skin wounds. A review. Am J Forensic Med Pathol 16:203–209

    Article  PubMed  CAS  Google Scholar 

  22. Martin P (1997) Wound healing—aiming for perfect skin regeneration. Science 276:75–81

    Article  PubMed  CAS  Google Scholar 

  23. Singer AJ, Clark RA (1999) Cutaneous wound healing. N Engl J Med 341:738–746

    Article  PubMed  CAS  Google Scholar 

  24. Gillitzer R, Goebeler M (2001) Chemokines in cutaneous wound healing. J Leukoc Biol 69:513–521

    PubMed  CAS  Google Scholar 

  25. Martin P, Leibovich SJ (2005) Inflammatory cells during wound repair: the good, the bad and the ugly. Trends Cell Biol 15:599–607

    Article  PubMed  CAS  Google Scholar 

  26. Ishida Y, Gao JL, Murphy PM (2008) Chemokine receptor CX3CR1 mediates skin wound healing by promoting macrophage and fibroblast accumulation and function. J Immunol 180:569–579

    PubMed  CAS  Google Scholar 

  27. Ishida Y, Kondo T, Kimura A, Matsushima K, Mukaida N (2006) Absence of IL-1 receptor antagonist impaired wound healing along with aberrant NF-κB activation and a reciprocal suppression of TGF-β signal pathway. J Immunol 176:5598–5606

    PubMed  CAS  Google Scholar 

  28. Ishida Y, Kondo T, Takayasu T, Iwakura Y, Mukaida N (2004) The essential involvement of cross-talk between IFN-γ and TGF-β in the skin wound-healing process. J Immunol 172:1848-1855

    Google Scholar 

  29. Lin ZQ, Kondo T, Ishida Y, Takayasu T, Mukaida N (2003) Essential involvement of IL-6 in the skin wound-healing process as evidenced by delayed wound healing in IL-6-deficient mice. J Leukoc Biol 73:713–721

    Article  PubMed  CAS  Google Scholar 

  30. Mori R, Kondo T, Ohshima T, Ishida Y, Mukaida N (2002) Accelerated wound healing in tumor necrosis factor receptor p55-deficient mice with reduced leukocyte infiltration. FASEB J 16:963–974

    Article  PubMed  CAS  Google Scholar 

  31. Laulederkind SJ, Thompson-Jaeger S, Goorha S, Chen Q, Fu A, Rho JY, Ballou LR, Raghow R (2002) Both constitutive and inducible prostaglandin H synthase affect dermal wound healing in mice. Lab Invest 82:919–927

    PubMed  CAS  Google Scholar 

  32. Dubois RN, Abramson SB, Crofford L, Gupta RA, Simon LS, Van De Putte LB, Lipsky PE (1998) Cyclooxygenase in biology and disease. FASEB J 12:1063–1073

    PubMed  CAS  Google Scholar 

  33. Warner TD, Mitchell JA (2004) Cyclooxygenases: new forms, new inhibitors, and lessons from the clinic. FASEB J 18:790–804

    Article  PubMed  CAS  Google Scholar 

  34. Foitzik T, Hotz HG, Hotz B, Wittig F, Buhr HJ (2003) Selective inhibition of cyclooxygenase-2 (COX-2) reduces prostaglandin E2 production and attenuates systemic disease sequelae in experimental pancreatitis. Hepatogastroenterology 50:1159–1162

    PubMed  CAS  Google Scholar 

  35. Hayashi T, Ishida Y, Kimura A, Iwakura Y, Mukaida N, Kondo T (2007) IFN-γ protects cerulein-induced acute pancreatitis by repressing NF-κB activation. J Immunol 178:7385–7394

    PubMed  CAS  Google Scholar 

  36. King SJ, Werrett DJ, Harrison DT (1989) The evaluation of an ELISA for semen-specific 19-OH prostaglandin F1 alpha/F2 alpha using ex-casework swabs and stains. Forensic Sci Int 42:103–123

    Article  PubMed  CAS  Google Scholar 

  37. King SJ, Kelly RW, Sutton JG (1989) The development of an enzyme-linked immunosorbent assay for 19-OH PG F1 alpha/F2 alpha. Forensic Sci Int 40:211–216

    Article  PubMed  CAS  Google Scholar 

  38. King SJ, Sutton JG, Trewsdale LA (1989) Radioimmunoassay detection limits for 19-OH F1 alpha/F2 alpha prostaglandin in normal, infertile and vasectomized semen stains. Analysis of saliva, sweat and urine for possible non-specific or matrix effects. Forensic Sci Int 40:221–229

    Article  PubMed  CAS  Google Scholar 

  39. King SJ, Sutton JG (1989) A survey of the concentration of 19-OH F1 alpha/F2 alpha prostaglandins in the semen of fertile, infertile and vasectomized men and their stability in both liquid semen and semen stains. Forensic Sci Int 40:217–220

    Article  PubMed  CAS  Google Scholar 

  40. Sutton JG, King SJ, Taylor M (1987) The application of a simple RIA technique for the detection of 19-OH F1 alpha/F2 alpha prostaglandin, a specific semen marker, in semen contaminated vaginal swabs: time since intercourse studies. Forensic Sci Int 34:143–153

    Article  PubMed  CAS  Google Scholar 

  41. Sutton JG, Kelly RW, Morris BA (1987) Evaluation of the 190H analogues of prostaglandins E1, E2, F1 alpha and F2 alpha as specific markers for the identification of human semen in body fluid mixtures. Forensic Sci Int 33:103–116

    Article  PubMed  CAS  Google Scholar 

  42. He L, Zhu J (1996) Distinguishing antemortem from postmortem injuries by LTB4 quantification. Forensic Sci Int 31:11–16

    Article  Google Scholar 

  43. Hernández-Cueto C, Vieira DN, Girela E, Marques E, Calvo MD, Villalobos M, Oliveira de Sà F, Villanueva E (1994) Prostaglandin F2a (PGF2a): an inadequate marker of the vitality of wounds? Int J Legal Med 106:312–314

    Article  PubMed  Google Scholar 

  44. Bai R, Wan L, Shi M (2008) The time-dependent expressions of IL-1beta, COX-2, MCP-1 mRNA in skin wounds of rabbits. Forensic Sci Int 175:193–197

    Article  PubMed  CAS  Google Scholar 

  45. Kondo T, Ohshima T (1996) The dynamics of inflammatory cytokines in the healing process of mouse skin wound: a preliminary study for possible wound age determination. Int J Legal Med 108:231–236

    Article  PubMed  CAS  Google Scholar 

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Acknowledgments

This study was financially supported by Grants-in-Aid for Scientific Research (A), Young Scientists (A), and Exploratory Research from the Ministry of Education, Science, Sports and Culture of Japan. We sincerely thank Ms. Mariko Kawaguchi for her excellent assistance in preparing this manuscript.

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Correspondence to Toshikazu Kondo.

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Ishida, Y., Kimura, A., Nosaka, M. et al. Immunohistochemical analysis on cyclooxygenase-2 for wound age determination. Int J Legal Med 126, 435–440 (2012). https://doi.org/10.1007/s00414-012-0685-7

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  • DOI: https://doi.org/10.1007/s00414-012-0685-7

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