title: Heparin-protamine does not aggravate local LPS-provoked leukocytic inflammation in vivo.
abstract: OBJECTIVE
  Secondary complications involving inflammation limit postoperative results in cardiac surgery. Because heparin-protamine can elicit inflammatory reactions, this study evaluates in vivo whether treatment with heparin-protamine aggravates local endotoxin-induced injury.
  
  
  METHODS
  Mice received intravenous injections of either heparin-protamine, protamine alone or PBS for controls, before local air pouch challenge with LPS. Leukocytes recruited within the air pouches were collected and analyzed by flow cytometry.
  
  
  RESULTS
  LPS provoked a local leukocytic infiltration in a dose- and time-dependent manner with significantly elevated numbers of 1.75 +/- 0.29 x 10 (6) cells after four hours compared to non-LPS-stimulated controls (0.55 +/- 0.08 x 10 (6) cells). Recruited cells comprised of 74 +/- 4 % PMNLs and 26 +/- 4 % MNLs. The largest fraction of MNLs was positive for the T cell-specific marker CD90.2 (59 +/- 6 %). B cells were only rarely observed (4 +/- 1 %). In non-LPS-challenged air pouches, heparin-protamine provoked a leukocytic infiltration, which was comparable to that observed after LPS (1.51 +/- 0.22 x 10 (6) cells). However, neither heparin-protamine nor protamine alone aggravated the LPS-mediated leukocyte recruitment (2.25 +/- 0.25 x 10 (6) and 1.77 +/- 0.23 x 10 (6) cells). Neither treatment influenced the distribution of leukocyte subpopulations compared to PBS-treated controls. Furthermore, surface expression of CD11a and CD11b on blood leukocytes did not differ between the groups, indicating that protamine does not increase the activation of circulating leukocytes during LPS-induced local inflammation.
  
  
  CONCLUSIONS
  Our data indicate that heparin-protamine, although pro-inflammatory in nature, does not aggravate local inflammation provoked by LPS. Thus, enhanced inflammation during the perioperative course of cardiac surgery patients seems not to be attributable to the intraoperative use of heparin-protamine.
authors.affiliation: Department of Thoracic and Cardiovascular Surgery, University of Saarland, Homburg/Saar, Germany. reneschramm@hotmail.com
authors.forename: R
authors.initials: R
authors.lastname: Schramm
authors.forename: R M
authors.initials: RM
authors.lastname: Nickels
authors.forename: Y
authors.initials: Y
authors.lastname: Harder
authors.forename: F
authors.initials: F
authors.lastname: Langer
authors.forename: M D
authors.initials: MD
authors.lastname: Menger
authors.forename: H-J
authors.initials: HJ
authors.lastname: Schäfers
mesh_terms: D000818:Animals; D000925:Anticoagulants / Q000494:pharmacology*; D005434:Flow Cytometry; D006493:Heparin / Q000494:pharmacology*; D006494:Heparin Antagonists / Q000494:pharmacology*; D007249:Inflammation; D007962:Leukocytes / Q000187:drug effects / Q000276:immunology / Q000473:pathology*; D008070:Lipopolysaccharides / Q000494:pharmacology; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D011479:Protamines / Q000494:pharmacology*
chemical_list: D000925:Anticoagulants; D006494:Heparin Antagonists; D008070:Lipopolysaccharides; D011479:Protamines; D006493:Heparin