Abstract
This study explored the impact of gold nanoparticles on the metabolic activity and morphology of human pulmonary endothelial cell monolayers. We developed a gold nanoparticle library of three different sizes and two surface chemistries that include anionic citrate and the cationic polyelectrolyte poly(allylamine hydrochloride). The nanoparticles were characterized in cell culture medium to assess how their physical properties are altered after exposure to biological fluids. A bovine serum albumin pretreatment protocol was developed to stabilize the nanoparticles in cell culture medium. Results of this study show that an 18 h exposure of human pulmonary artery endothelial cells to the different nanoparticles modestly affects cellular metabolic activity. However, nanoparticle exposure perturbs the cortical actin networks and induces the formation of intercellular gaps. In particular, exposure to the poly(allylamine hydrochloride)-coated particles reduces the area of cell–cell junctions—a change that correlates with increased leakiness of endothelial barriers. The presence of excess polyelectrolyte capping agents in the supernatant of poly(allylamine hydrochloride)-coated nanoparticles significantly impacts endothelial morphology. Pretreatment of the particle supernatant with bovine serum albumin mitigates the negative effects of free or bound polyelectrolytes on endothelial cell monolayers.
Highlights
Exposure to biological fluids alters the physical properties of NPs, due to the adsorption of proteins and other biological macromolecules present in biological fluids[24]
To reveal the environmental impact of NPs on the human lung, we report the acute impact of gold nanoparticles (AuNPs) on the metabolic activity of primary, human pulmonary artery endothelial cells (HPAECs) and on the endothelial monolayer integrity
We compared the effects of bovine serum albumin (BSA) pretreatment of NPs on endothelial cell metabolic activity and morphology
Summary
Exposure to biological fluids alters the physical properties of NPs, due to the adsorption of proteins and other biological macromolecules present in biological fluids[24]. The NP aggregation state can be altered in the presence of biomolecules – for instance, proteins have been found to stabilize colloidal NPs, while protein-free biological fluids induce NP a ggregation[25]. While coating NPs with a protein corona can lead to off target or undesired biological responses, Chen et al were the first to develop a pre-formed protein corona to achieve safe biomedical and environmental effects of ZnO N Ps27–29. Developing processing protocols for maintaining NP colloidal stability in complex biological media is desirable. Outlining a bovine serum albumin (BSA) pretreatment protocol, we developed pre-formed protein coronas to stabilize AuNPs in biological media. This study compares the effects of NP surface coating, size, and dose on the metabolic activity, cytoskeletal organization, and morphology of pulmonary endothelial cells
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