Effect of natural porcine surfactant in Staphylococcus aureus induced pro-inflammatory cytokines and reactive oxygen species generation in monocytes and neutrophils from human blood

Abstract

Surfacen (R) is a clinical surfactant preparation of porcine origin. in the present study, we have evaluated the effect of Surfacen (R) in the modulation of oxidative burst in monocytes and neutrophils in human blood and proinflammatory cytokine production in peripheral blood mononuclear cells (PBMC). Reactive oxygen species (ROS) level was measured in monocytes and neutrophils by flow cytometry using 2,7-dichlorofluorescein diacetate (DCFH-DA) as substrate, while, tumor necrosis factor (TNF)-alpha and interleukin (IL)-6 levels were estimated in PBMC supernatant by enzyme-linked immunosorbent assays (ELISA). Our results show that Staphylococcus aureus-induced ROS level was slightly affected by Surfacen (R) added to whole blood monocytes and neutrophils. the time course experiments of pre-incubation with Surfacen (R) showed no significant increase of ROS level at 2 h; however, the ROS level decreased when pre incubated for 4 h and 6 h with Surfacen (R). Pre-incubation of PBMC cells with Surfacen (R) at 0.125 and 0.5 mg/mL showed a dose-dependent suppression of TNF-alpha levels measured after 4 h of S. aureus stimulation, an effect less impressive when cells were stimulated for 24 h. A similar behavior was observed in IL-6 release. in summary, the present study provides experimental evidence supporting an anti-inflammatory role of Surfacen (R) in human monocytes and neutrophils in vitro. (C) 2014 Elsevier B.V. All rights reserved.