title: In vitro impairment of whole blood coagulation and platelet function by hypertonic saline hydroxyethyl starch.
abstract: BACKGROUND
  Hypertonic saline hydroxyethyl starch (HH) has been recommended for first line treatment of hemorrhagic shock. Its effects on coagulation are unclear. We studied in vitro effects of HH dilution on whole blood coagulation and platelet function. Furthermore 7.2% hypertonic saline, 6% hydroxyethylstarch (as ingredients of HH), and 0.9% saline solution (as control) were tested in comparable dilutions to estimate specific component effects of HH on coagulation.
  
  
  METHODS
  The study was designed as experimental non-randomized comparative in vitro study. Following institutional review board approval and informed consent blood samples were taken from 10 healthy volunteers and diluted in vitro with either HH (HyperHaes, Fresenius Kabi, Germany), hypertonic saline (HT, 7.2% NaCl), hydroxyethylstarch (HS, HAES6%, Fresenius Kabi, Germany) or NaCl 0.9% (ISO) in a proportion of 5%, 10%, 20% and 40%. Coagulation was studied in whole blood by rotation thrombelastometry (ROTEM) after thromboplastin activation without (ExTEM) and with inhibition of thrombocyte function by cytochalasin D (FibTEM), the latter was performed to determine fibrin polymerisation alone. Values are expressed as maximal clot firmness (MCF, [mm]) and clotting time (CT, [s]). Platelet aggregation was determined by impedance aggregrometry (Multiplate) after activation with thrombin receptor-activating peptide 6 (TRAP) and quantified by the area under the aggregation curve (AUC [aggregation units (AU)/min]). Scanning electron microscopy was performed to evaluate HyperHaes induced cell shape changes of thrombocytes.
  
  
  METHODS
  2-way ANOVA for repeated measurements, Bonferroni post hoc test, p < 0.01.
  
  
  RESULTS
  Dilution impaired whole blood coagulation and thrombocyte aggregation in all dilutions in a dose dependent fashion. In contrast to dilution with ISO and HS, respectively, dilution with HH as well as HT almost abolished coagulation (MCFExTEM from 57.3 ± 4.9 mm (native) to 1.7 ± 2.2 mm (HH 40% dilution; p < 0.0001) and to 6.6 ± 3.4 mm (HT 40% dilution; p < 0.0001) and thrombocyte aggregation (AUC from 1067 ± 234 AU/mm (native) to 14.5 ± 12.5 AU/mm (HH 40% dilution; p < 0.0001) and to 20.4 ± 10.4 AU/min (HT 40% dilution; p < 0.0001) without differences between HH and HT (MCF: p = 0.452; AUC: p = 0.449).
  
  
  CONCLUSIONS
  HH impairs platelet function during in vitro dilution already at 5% dilution. Impairment of whole blood coagulation is significant after 10% dilution or more. This effect can be pinpointed to the platelet function impairing hypertonic saline component and to a lesser extend to fibrin polymerization inhibition by the colloid component or dilution effects.Accordingly, repeated administration and overdosage should be avoided.
authors.affiliation: Department of Anaesthesiology and Intensive Care Medicine, Hannover Medical School, Germany. hanke.alexander@mh-hannover.de
authors.forename: Alexander A
authors.initials: AA
authors.lastname: Hanke
authors.forename: Stephanie
authors.initials: S
authors.lastname: Maschler
authors.forename: Herbert
authors.initials: H
authors.lastname: Schöchl
authors.forename: Felix
authors.initials: F
authors.lastname: Flöricke
authors.forename: Klaus
authors.initials: K
authors.lastname: Görlinger
authors.forename: Klaus
authors.initials: K
authors.lastname: Zanger
authors.forename: Peter
authors.initials: P
authors.lastname: Kienbaum
mesh_terms: D000328:Adult; D001777:Blood Coagulation / Q000187:drug effects*; D001780:Blood Coagulation Tests; D001792:Blood Platelets / Q000187:drug effects*; D005260:Female; D006801:Humans; D006895:Hydroxyethyl Starch Derivatives / Q000494:pharmacology*; D008297:Male; D010952:Plasma Substitutes / Q000494:pharmacology; D012462:Saline Solution, Hypertonic / Q000494:pharmacology*
chemical_list: D006895:Hydroxyethyl Starch Derivatives; D010952:Plasma Substitutes; D012462:Saline Solution, Hypertonic