Hypertonic saline, mannitol and hydroxyethyl starch preconditioning of platelets obtained from septic patients attenuates CD40 ligand expression in vitro

Background: Because platelet CD40 ligand (CD40L) expression plays an important role in inflammatory conditions, reduction of CD40L expression may be beneficial for patients with sepsis. Although hypertonic saline, mannitol, and hydroxyethyl starch (HES) solutions have been shown to modulate inflammatory responses, their effects on platelet CD40L expression are unclear. We assessed the effects of hypertonic saline, mannitol, and HES solutions on platelet CD40L expression. Methods: Platelet-rich plasma samples were obtained from septic patients and diluted to 1%, 2.5%, 5%, or 7.5% (vol/vol) with 7.5% saline, 3% saline, 0.9% saline, 20% mannitol, 10% HES (200/0.5), or Ringer's solution. Twenty-five samples were used per dilution. To determine platelet CD40L expression, platelet samples were stimulated with thrombin (0.1 U/mL), incubated with fluorochrome-conjugated platelet antibodies, and analyzed using flow cytometry. Results: Preconditioning of platelet-rich plasma with hypertonic saline, mannitol, and HES attenuated CD40L expression at dilution ratios of 5%, 1%, and 1%, respectively. The decreases were concentration dependent. The effects of mannitol and HES on CD40L expression were almost identical and were superior to those of 3% saline. In contrast, 0.9% saline and Ringer's solution had no effect on CD40L expression. Conclusions: Our data show that resuscitation fluids, such as hypertonic saline, mannitol, and HES, inhibit agonist-induced CD40L expression on platelets. These resuscitation fluids may have an anti-inflammatory action when administered to septic patients.