title: Polymicrobial sepsis induces organ changes due to granulocyte adhesion in a murine two hit model of trauma.
abstract: BACKGROUND
  Polytrauma patients, who develop organ dysfunction, have often undergone multiple subsequent insults ("hits"). The sequence of organs that show a dysfunction mostly is lung, liver, kidney and heart. The aim of the present study was to investigate whether a second hit after trauma induces organ changes. Furthermore, it was of interest to identify possible pathogenic mediators such as polymorphonuclear granulocytes (PMN) and cytokines. For this purpose, a two hit model of systemic damage in mice was developed. Sepsis was induced by caecal ligation and puncture (CLP), which was preceded 48 hours by a femur fracture, the most common fracture of long bones in trauma patients. This fracture was combined with a haemorrhagic shock.
  
  
  METHODS
  In both mouse groups studied, a standardized femur fracture was produced using a blunt guillotine device with a weight of 500 g. This was followed by a haemorrhagic shock with substitution of ringer's lactate after 1 hour. In the study group, CLP was induced by puncturing the caecum using a 21G needle. As a control, sham animals underwent a laparotomy without CLP. Both groups were sacrificed after 48 or 96 hours. Clinical parameters were investigated on a daily basis to evaluate the animals' status. Lung, liver and kidney morphology was studied by light microscopy. PMN adhesion was determined by counting the number of adherent PMN per 100 microm of endothelium. Serum levels of TNF-alpha were measured after 48 and 96 hours.
  
  
  RESULTS
  In the group submitted to laparotomy, all animals survived. The induction of polymicrobial sepsis by CLP resulted in an 85% (34/40) mortality within 96 hours after surgery (p < 0.05). The induction of a polymicrobial sepsis resulted in a significantly steady worsening of the clinical situation compared to the sham animals (p < 0.05). Lung morphology demonstrated significant changes at the end of the experimental period after 96 h in the two hit group. The alveolar septa were thickened and in all lungs haemorrhagic foci were observed. The number of PMN adhering to the pulmonary endothelium significantly increased at 96 hours. Some of the liver specimens in the two hit group showed focal hydropic degeneration and PMN infiltration. No kidney pathology was observed. This result coincided with an increase in TNF-alpha serum levels.
  
  
  CONCLUSIONS
  A new rodent model mimicking the situation in the polytraumatized patient was developed. Although the animals showed minimal organ manifestation, a high percentage died probably due to cytokinemia. Furthermore, the increased TNF-alpha levels may lead to increased adhesion of PMN in the lung venules. This adhesion developed four days after the second hit. This might be the initial step for the development of extensive lung lesions in later phases. This model represents the SIRS more than MODS. This is a model for devolopment of posttraumatic disease due to cytokinemia and less for chronic multiple organ dysfunction and failure.
authors.affiliation: Departments of 'Trauma Surgery, Hannover Medical School, Hannover, Germany. Griensven.Martijn.van@MH-Hannover.de
authors.forename: Martijn
authors.initials: M
authors.lastname: van Griensven
authors.forename: Meryem
authors.initials: M
authors.lastname: Kuzu
authors.forename: Maike
authors.initials: M
authors.lastname: Breddin
authors.forename: Frederic
authors.initials: F
authors.lastname: Böttcher
authors.forename: Christian
authors.initials: C
authors.lastname: Krettek
authors.forename: Hans-Christoph
authors.initials: HC
authors.lastname: Pape
authors.forename: Thomas
authors.initials: T
authors.lastname: Tschernig
mesh_terms: D000818:Animals; D002448:Cell Adhesion / Q000502:physiology; D005264:Femoral Fractures / Q000150:complications* / Q000382:microbiology / Q000503:physiopathology; D007668:Kidney / Q000473:pathology; D008099:Liver / Q000473:pathology; D008168:Lung / Q000473:pathology*; D008297:Male; D051379:Mice; D008815:Mice, Inbred Strains; D009504:Neutrophils / Q000166:cytology*; D018805:Sepsis / Q000150:complications / Q000382:microbiology* / Q000503:physiopathology