title: A new method to determine wound age in early vital skin injuries: a probability scoring system using expression levels of Fibronectin, CD62p and Factor VIII in wound hemorrhage.
abstract: OBJECTIVE
  In forensic autopsies it is important to determine the age of early vital skin wounds as accurate as possible. In addition to inflammation, coagulation is also induced in vital wounds. Analysis of blood coagulation markers in wound hemorrhage could therefore be an important additional discriminating factor in wound age determination. The aim of this study was to develop a wound age probability scoring system, based on the immunohistochemical expression levels of Fibronectin, CD62p and Factor VIII in wound hemorrhage.
  
  
  METHODS
  Tissue samples of (A) non injured control skin (n=383), and samples of mechanically induced skin injuries of known wound age, (B) injuries inflicted shortly before death (up to a few minutes old) (n=382), and (C) injuries inflicted 15-30 min before death (n=42) were obtained at autopsy in order to validate wound age estimation. Tissue slides were stained for Fibronectin, CD62p and Factor VIII and were subsequently scored for staining intensity (IHC score) in wound hemorrhage (1=minor, 2=moderate, 3=strong positive). Finally, probability scores of these markers were calculated.
  
  
  RESULTS
  In at most 14% of the non-injured control samples, hemorrhage was found, with mean±standard deviation IHC scores of 0.1±0.4, 0.2±0.4 and 0.2±0.5 for Fibronectin, CD62p, and Factor VIII, respectively. Expression of these markers significantly increased to mean IHC scores 1.4±0.8 (Fibronectin), 1.2±0.6 (CD62p), and 1.6±0.8 (Factor VIII) in wounds inflicted shortly before death (few minutes old) and to 2.6±0.5 (Fibronectin), 2.5±0.6 (CD62p), and 2.8±0.4 (Factor VIII) in 15-30 min old wounds. The probabilities that a wound was non vital in case of an IH score 0 were 87%, 88% and 90% for Fibronectin, CD62p, and Factor VIII, respectively. In case of an IHC score 1 or 2, the probabilities that a wound was a few minutes old were 82/90%, 82/83% and 72/93%. Finally, in case of an IHC score 3, the probabilities that a wound was 15-30 min old were 65%, 76% and 55%.
  
  
  CONCLUSIONS
  Based on the expression of Fibronectin, CD62p and Factor VIII in wound hemorrhage, we developed a probability scoring system that can be used in forensic autopsies to improve wound age estimation in early skin injuries.
authors.affiliation: Netherlands Forensic Institute (NFI), The Hague, The Netherlands; Department of Pathology, VU University Medical Centre, 1007 HV Amsterdam, The Netherlands.
authors.forename: Franklin R W
authors.initials: FR
authors.lastname: van de Goot
authors.affiliation: Department of Pathology, VU University Medical Centre, 1007 HV Amsterdam, The Netherlands; ICaR-VU, VU University Medical Centre, Amsterdam, The Netherlands. Electronic address: h.korkmaz@vumc.nl.
authors.forename: H Ibrahim
authors.initials: HI
authors.lastname: Korkmaz
authors.affiliation: Department of Pathology, VU University Medical Centre, 1007 HV Amsterdam, The Netherlands; Symbiant Pathology Expert Centre, Alkmaar, The Netherlands.
authors.forename: Judith
authors.initials: J
authors.lastname: Fronczek
authors.affiliation: Department of Epidemiology and Statistics, VU University Medical Centre, Amsterdam, The Netherlands.
authors.forename: Birgit I
authors.initials: BI
authors.lastname: Witte
authors.affiliation: Netherlands Forensic Institute (NFI), The Hague, The Netherlands.
authors.forename: Rob
authors.initials: R
authors.lastname: Visser
authors.affiliation: Department of Molecular Cell Biology and Immunology, VU University Medical Centre, Amsterdam, The Netherlands; MOVE Research Institute Amsterdam, VU University Medical Centre, Amsterdam, The Netherlands; Association of Dutch Burn Centres (ADBC), Beverwijk, The Netherlands.
authors.forename: Magda M W
authors.initials: MM
authors.lastname: Ulrich
authors.affiliation: Netherlands Forensic Institute (NFI), The Hague, The Netherlands; Department of Pathology, VU University Medical Centre, 1007 HV Amsterdam, The Netherlands; ICaR-VU, VU University Medical Centre, Amsterdam, The Netherlands.
authors.forename: Mark P V
authors.initials: MP
authors.lastname: Begieneman
authors.affiliation: Department of Pathology, VU University Medical Centre, 1007 HV Amsterdam, The Netherlands.
authors.forename: Lawrence
authors.initials: L
authors.lastname: Rozendaal
authors.affiliation: Department of Pathology, VU University Medical Centre, 1007 HV Amsterdam, The Netherlands; ICaR-VU, VU University Medical Centre, Amsterdam, The Netherlands.
authors.forename: Paul A J
authors.initials: PA
authors.lastname: Krijnen
authors.affiliation: Department of Pathology, VU University Medical Centre, 1007 HV Amsterdam, The Netherlands; Department of Cardiac Surgery, VU University Medical Centre, Amsterdam, The Netherlands; ICaR-VU, VU University Medical Centre, Amsterdam, The Netherlands.
authors.forename: Hans W M
authors.initials: HW
authors.lastname: Niessen
mesh_terms: D000293:Adolescent; D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D015415:Biomarkers / Q000378:metabolism; D016022:Case-Control Studies; D002648:Child; D002675:Child, Preschool; D005169:Factor VIII / Q000378:metabolism*; D005353:Fibronectins / Q000378:metabolism*; D049429:Forensic Pathology / Q000379:methods; D006470:Hemorrhage / Q000378:metabolism* / Q000473:pathology; D006801:Humans; D007150:Immunohistochemistry; D007223:Infant; D007231:Infant, Newborn; D008875:Middle Aged; D019007:P-Selectin / Q000378:metabolism*; D012867:Skin / Q000293:injuries / Q000378:metabolism* / Q000473:pathology; D013997:Time Factors; D014949:Wounds, Nonpenetrating / Q000378:metabolism / Q000473:pathology; D055815:Young Adult
chemical_list: D015415:Biomarkers; D005353:Fibronectins; D019007:P-Selectin; C546937:SELP protein, human; D005169:Factor VIII
keywords: Autopsy; Forensic; Immunohistochemistry; Immunology; Skin wounds; Wound age determination