title: Analysis of morphological characteristics and expression levels of extracellular matrix proteins in skin wounds to determine wound age in living subjects in forensic medicine.
abstract: OBJECTIVE
  Wound age determination in living subjects is important in routine diagnostics in forensic medicine. Macroscopical description of a wound to determine wound age however is inadequate. The aim of this study was to assess whether it would be feasible to determine wound age via analysis of morphological characteristics and extracellular matrix proteins in skin biopsies of living subjects referred to a forensic outpatient clinic.
  
  
  METHODS
  Skin biopsies (n=101), representing the border area of the wound, were taken from skin injuries of known wound age (range: 4.5h-25 days) in living subjects. All biopsies were analyzed for 3 morphological features (ulceration, parakeratosis and hemorrhage) and 3 extracellular matrix markers (collagen III, collagen IV and α-SMA). For quantification, biopsies were subdivided in 4 different timeframes: 0.2-2 days, 2-4 days, 4-10 days and 10-25 days old wounds. Subsequently, a probability scoring system was developed.
  
  
  RESULTS
  For hemorrhage, collagen III, collagen IV and α-SMA expression no relation with wound age was found. Ulceration was only found in wounds of 0.2-2, 2-4 and 4-10 days old, implying that the probability that a wound was more than 10 days old in case of ulceration is equal to 0%. Also parakeratosis was almost exclusively found in wounds of 0.2-2, 2-4 and 4-10 days old, except for one case with a wound age of 15 days old. The probability scoring system of all analyzed markers, as depicted above, however can be used to calculate individual wound age probabilities in biopsies of skin wounds of living subjects.
  
  
  CONCLUSIONS
  We have developed a probability scoring system, analysing morphological characteristics and extracellular matrix proteins in superficial skin biopsies of wounds in living subjects that can be applied in forensic medicine for wound age determination.
authors.affiliation: Department of Pathology, VU Medical Centre, De Boelelaan 1117, 1081 HV Amsterdam, The Netherlands; Department of Pathology, Symbiant, Medisch Centrum Alkmaar, Wilhelminalaan 12, 1815 JD Alkmaar, The Netherlands.
authors.forename: Judith
authors.initials: J
authors.lastname: Fronczek
authors.affiliation: Public Health Service Amsterdam, Department of Forensic Medicine, Nieuwe Achtergracht 100, 1018 WT Amsterdam, The Netherlands.
authors.forename: Ronald
authors.initials: R
authors.lastname: Lulf
authors.affiliation: Department of Pathology, VU Medical Centre, De Boelelaan 1117, 1081 HV Amsterdam, The Netherlands; ICaR-VU, VU Medical Centre, De Boelelaan 1117, 1081 HV Amsterdam, The Netherlands. Electronic address: h.korkmaz@vumc.nl.
authors.forename: H Ibrahim
authors.initials: HI
authors.lastname: Korkmaz
authors.affiliation: Department of Epidemiology and Biostatistics, VU Medical Centre, De Boelelaan 1117, 1081 HV Amsterdam, The Netherlands.
authors.forename: Birgit I
authors.initials: BI
authors.lastname: Witte
authors.affiliation: Department of Pathology, Symbiant, Medisch Centrum Alkmaar, Wilhelminalaan 12, 1815 JD Alkmaar, The Netherlands.
authors.forename: Franklin R W
authors.initials: FR
authors.lastname: van de Goot
authors.affiliation: Department of Pathology, VU Medical Centre, De Boelelaan 1117, 1081 HV Amsterdam, The Netherlands; ICaR-VU, VU Medical Centre, De Boelelaan 1117, 1081 HV Amsterdam, The Netherlands; Department of Pathology, Nederlands Forensisch Instituut (NFI), Laan van Ypenburg 6, 2490 GB The Hague, The Netherlands.
authors.forename: Mark P V
authors.initials: MP
authors.lastname: Begieneman
authors.affiliation: Department of Pathology, VU Medical Centre, De Boelelaan 1117, 1081 HV Amsterdam, The Netherlands; ICaR-VU, VU Medical Centre, De Boelelaan 1117, 1081 HV Amsterdam, The Netherlands.
authors.forename: Paul A J
authors.initials: PA
authors.lastname: Krijnen
authors.affiliation: Department of Pathology, VU Medical Centre, De Boelelaan 1117, 1081 HV Amsterdam, The Netherlands.
authors.forename: Lawrence
authors.initials: L
authors.lastname: Rozendaal
authors.affiliation: Department of Pathology, VU Medical Centre, De Boelelaan 1117, 1081 HV Amsterdam, The Netherlands; ICaR-VU, VU Medical Centre, De Boelelaan 1117, 1081 HV Amsterdam, The Netherlands; Department of Cardiac Surgery, VU Medical Centre, De Boelelaan 1117, 1081 HV Amsterdam, The Netherlands.
authors.forename: Hans W M
authors.initials: HW
authors.lastname: Niessen
authors.affiliation: Public Health Service Amsterdam, Department of Forensic Medicine, Nieuwe Achtergracht 100, 1018 WT Amsterdam, The Netherlands.
authors.forename: Udo J L
authors.initials: UJ
authors.lastname: Reijnders
mesh_terms: D000199:Actins / Q000378:metabolism; D000293:Adolescent; D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D001706:Biopsy; D024061:Collagen Type III / Q000378:metabolism; D024141:Collagen Type IV / Q000378:metabolism; D005260:Female; D049429:Forensic Pathology; D006470:Hemorrhage / Q000378:metabolism / Q000473:pathology; D006801:Humans; D007150:Immunohistochemistry; D007642:Keratosis / Q000378:metabolism / Q000473:pathology; D008297:Male; D008875:Middle Aged; D058628:Myofibroblasts / Q000378:metabolism; D012867:Skin / Q000293:injuries* / Q000378:metabolism* / Q000473:pathology; D012883:Skin Ulcer / Q000378:metabolism / Q000473:pathology; D013997:Time Factors; D014945:Wound Healing / Q000502:physiology*; D055815:Young Adult
chemical_list: C541116:ACTA2 protein, human; D000199:Actins; D024061:Collagen Type III; D024141:Collagen Type IV
keywords: Forensic; Immunohistochemistry; Immunology; Living subjects; Skin wounds; Wound age determination