Novel electrospun chitosan/PEO membranes for more predictive nanoparticle transport studies at biological barriers.

The design of safe and effective nanoparticles (NPs) for commercial and medical applications requires a profound understanding of NP translocation and effects at biological barriers. To gain mechanistic insights, physiologically relevant and accurate human in vitro biobarrier models are indispensable. However, current transfer models largely rely on artificial porous polymer membranes for the cultivation of cells, which do not provide a close mimic of the natural basal membrane and intrinsically provide limited permeability for NPs. In this study, electrospinning is exploited to develop thin chitosan/polyethylene oxide (PEO) membranes with a high porosity and nanofibrous morphology for more predictive NP transfer studies. The nanofiber membranes allow the cultivation of a tight and functional placental monolayer (BeWo trophoblasts). Translocation studies with differently sized molecules and NPs (Na-fluorescein; 40 kDa FITC-Dextran; 25 nm PMMA; 70, 180 and 520 nm polystyrene NPs) across empty and cell containing membranes reveal a considerably enhanced permeability compared to commercial microporous membranes. Importantly, the transfer data of NPs is highly similar to data from ex vivo perfusion studies of intact human placental tissue. Therefore, the newly developed membranes may decisively contribute to establish physiologically relevant in vitro biobarrier transfer models with superior permeability for a wide range of molecules and particles.

Airborne emissions from combustion of graphene nanoplatelet/epoxy composites and their cytotoxicity on lung cells via air-liquid interface cell exposure in vitro.

Graphene nanoplatelet (GNP) as a nanofiller improves the mechanical strength, electrical conductivity, and flame retardancy of the polymers significantly. With an increasing number of GNP-reinforced products, a careful safety assessment is needed to avoid social and economic setbacks. However, no study has addressed the effects of combustion-generated emissions from GNP-reinforced products in the lung, the most sensitive exposure route to airborne particles. Therefore, we studied the influence of GNP as a nanofiller on the emitted particles and polycyclic aromatic hydrocarbons (PAHs), and cytotoxicity of the emissions from the combustion of pure epoxy (EP) and GNP-reinforced epoxy (EP-GNP). GNP was not detected in the airborne emissions. PAHs were found in airborne particles of both emissions from EP and EP-GNP, with some differences in their concentrations. A first hazard assessment was performed on human alveolar epithelial cells exposed to the airborne emissions at air-liquid interface conditions. At 24 h and 96 h after the exposure, similar responses were observed between EP and EP-GNP except an acute transient decrease in mitochondrial activity after exposure to the emissions from EP-GNP. Both emissions from EP and EP-GNP had no acute effects on membrane integrity, cell morphology or expression of anti-oxidative stress markers (HMOX1 and SOD2 genes). Meanwhile, both emissions induced the activation of the aryl hydrocarbon receptor (CYP1A1 gene) and a transient (pro-) inflammatory response (MCP-1), but the effects between EP and EP-GNP were not significantly different.

Investigating the effects of differently produced synthetic amorphous silica (E 551) on the integrity and functionality of the human intestinal barrier using an advanced in vitro co-culture model.

E 551, also known as synthetic amorphous silica (SAS), is the second most produced food additive. However, according to the re-evaluation of E 551 by the European Food Safety Authority (EFSA) in 2018, the amount of available data on the oral toxicity of food grade E 551 is still insufficient for reliable risk assessment. To close this gap, this study aimed to investigate six food-grade SAS with distinct physicochemical properties on their interaction with the intestinal barrier using advanced in vitro intestinal co-cultures and to identify potential structure-activity relationships. A mucus-secreting Caco-2/HT-29/Raji co-culture model was treated with up to 50 µg/ml SAS for 48 h, which represents a dose range relevant to dietary exposure. No effects on cell viability, barrier integrity, microvilli function or the release of inflammatory cytokine were detected after acute exposure. Slight biological responses were observed for few SAS materials on iron uptake and gene expression levels of mucin 1 and G-protein coupled receptor 120 (GPR120). There was no clear correlation between SAS properties (single or combined) and the observed biological responses. Overall, this study provides novel insights into the short-term impact of food-relevant SAS with distinct characteristics on the intestinal epithelium including a range of intestine-specific functional endpoints. In addition, it highlights the importance of using advanced intestinal co-cultures embracing relevant cell types as well as a protective mucus barrier to achieve a comprehensive understanding of the biological response of food additives at the intestinal barrier in vitro.