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® is a clinical surfactant preparation of porcine origin. In the present study, we have evaluated the effect of Surfacen® in the modulation of oxidative burst in monocytes and neutrophils in human blood and pro-inflammatory 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)-α 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® added to whole blood monocytes and neutrophils. The time course experiments of pre-incubation with Surfacen® showed no significant increase of ROS level at 2h; however, the ROS level decreased when pre incubated for 4h and 6h with Surfacen®. Pre-incubation of PBMC cells with Surfacen® at 0.125 and 0.5mg/mL showed a dose-dependent suppression of TNF-α levels measured after 4h of S. aureus stimulation, an effect less impressive when cells were stimulated for 24h. A similar behavior was observed in IL-6 release. In summary, the present study provides experimental evidence supporting an anti-inflammatory role of Surfacen® in human monocytes and neutrophils in vitro.

Effects of the New Generation Synthetic Reconstituted Surfactant CHF5633 on Pro- and Anti-Inflammatory Cytokine Expression in Native and LPS-Stimulated Adult CD14+ Monocytes

“…Effects of CHF5633 on LPS-induced TNF-α and IL-1β expression paralleled those of Curosurf®. The latter results are in accordance with previous data from our group and with results from a number of other studies on natural surfactants such as Curosurf® [ 54 , 56 , 58 , 62 , 63 , 65 , 68 – 70 ]. Although described in previous studies [ 64 , 67 , 74 ], we did not find an equivalent anti-inflammatory activity when evaluating the major synthetic components of CHF5633 alone.…”

“…For natural surfactant preparations as well as some surfactant phospholipids, beneficial anti-inflammatory effects, such as surfactant-induced modulation of phagocyte function, modulation of oxidative burst, and suppression of pro-inflammatory cytokine release have been demonstrated in a relevant number of studies [ 54 – 70 ]. Data on potential immunomodulatory capacities of CHF5633 is limited, so far.…”

Staphylococcus aureus-mediated blood-brain barrier injury: anin vitrohuman brain microvascular endothelial cell model

“…Moreover, treatment of cultured HUVECs with SA has been reported to induce production and release of IL‐6 (Park et al, ; Soderquist et al, 2006), while elevated levels of IL‐6 have recently been reported in the plasma of patients suffering with Staphylococcal bloodstream infection (McNicholas et al, ). Evidence linking SA infection to ROS generation has also been reported in neutrophils, monocytes, macrophages, and resident bone marrow stem cells (de Guavara et al, ; Nandi et al, ). SA‐mediated ROS signaling in endothelial cells is, to our knowledge, limited to a recent paper demonstrating that SA‐purified LTA extract could induce permeabilization of pulmonary microvessel endothelial monolayers in a ROS‐dependent manner (Pai et al, ).…”

Activation of the AGE/RAGE system in the brains of rats and in SH-SY5Y cells exposed to high level of fluoride might connect to oxidative stress

“…Here, the DCFDA kit was used to measure hydroxyl, peroxyl, H 2 O 2 and other types of ROS in the cells, in the same manner as numerous recent investigations (Chang et al, 2014;Guo et al, 2014;de Guevara et al, 2014: Kastl et al, 2014Sato et al, 2014;Kuo et al, 2013), where the incubation time with various cell lines in vitro was variable from 30 min; 2, 6, 24 and 36 h; and even 7 days. However, since the half-life of ROS is quite short and the procedure detected by DCFDA kit reveals only the immediate situation, comprehensive consideration should be given when employing the method by DCFDA kit for detecting ROS.…”