title: Lack of plasminogen does not alter the early inflammatory response following a tympanic membrane perforation: a study in plasminogen-deficient mice.
abstract: CONCLUSIONS
  The results of the present study show that the early inflammatory response in plasminogen (plg)-deficient mice is not altered compared to that in wild-type (wt) mice. Therefore the chronicity of the perforation in the long-term healing experiment cannot be explained by an impairment of the early inflammatory response, but rather by an impairment in activation of the inflammatory cells. These findings give further insight into the mechanisms resulting in a clinically seen chronic tympanic membrane (TM) perforation and thus possible therapeutic strategies to replace today's conventional surgical treatment of these perforations.
  
  
  OBJECTIVE
  Plg has been shown to play an essential role in the healing of TM perforations. In plg-deficient mice a completely arrested healing reaction was seen, resulting in a chronic TM perforation. The mechanisms involved seem to be an abundant neutrophil recruitment, an accumulation of macrophages, an arrested keratinocyte migration, and a massive deposition of fibrin along the TM tissue. However, the exact functional role of plg in the early inflammatory response during healing of TM perforation remains unclear. This study aimed to evaluate the early inflammatory response, mainly the occurrence of macrophages and neutrophils, during the first 48 h following a perforation in the pars tensa (PT) of the TM, in mice lacking the plasminogen gene compared to the corresponding response in wt mice.
  
  
  METHODS
  The TMs were perforated in 45 plg-deficient and 39 wt mice. Otomicroscopic evaluation was performed at 3, 6, 9, 12, 18, 24, and 48 h after the perforation was made. Mice were harvested at all time points and prepared for morphology including immunohistochemistry (IHC). IHC was performed with antibodies targeting macrophages, neutrophils, T and B cells, cytokeratin, and fibrin(ogen). Morphometry was performed regarding the volume percentage of TM tissue occupied by the different inflammatory cells.
  
  
  RESULTS
  Perforation of the TM resulted in early otomicroscopic changes of the pars flaccida (PF) in both genotypes. Infiltration of inflammatory cells to PF and the presence of edema occurred as early as 6 h after the perforation was made, in both plg-deficient and wt mice. Morphometry did not reveal any significant differences between the genotypes concerning the occurrence of inflammatory cells. In contrast to the PF, the PT showed only sparse reactions during the experimental period. Furthermore, the migration pattern of keratinocytes did not differ between the genotypes throughout the experimental period.
authors.affiliation: Department of Clinical Sciences, Otorhinolaryngology, Umeå University, Umeå, Sweden. annika.hansson@ent.umu.se
authors.forename: Annika Hansson
authors.initials: AH
authors.lastname: Prestwich
authors.forename: Jinan
authors.initials: J
authors.lastname: Li
authors.forename: P O
authors.initials: PO
authors.lastname: Eriksson
authors.forename: Tor
authors.initials: T
authors.lastname: Ny
authors.forename: Diana
authors.initials: D
authors.lastname: Berggren
authors.forename: Sten
authors.initials: S
authors.lastname: Hellström
mesh_terms: D000818:Animals; D005838:Genotype; D007249:Inflammation; D008214:Lymphocytes / Q000502:physiology*; D008264:Macrophages / Q000502:physiology*; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D008811:Mice, Inbred DBA; D008853:Microscopy; D020556:Neutrophil Infiltration; D009504:Neutrophils / Q000502:physiology*; D010958:Plasminogen / Q000172:deficiency*; D014432:Tympanic Membrane / Q000473:pathology; D018058:Tympanic Membrane Perforation / Q000276:immunology / Q000473:pathology / Q000503:physiopathology*
chemical_list: D010958:Plasminogen