Abstract
As a highly interdisciplinary field, working with nanoparticles in a biomedical context requires a robust understanding of soft matter physics, colloidal behaviors, nano-characterization methods, biology, and bio-nano interactions. When reporting results, it can be easy to overlook simple, seemingly trivial experimental details. In this context, we set out to understand how in vitro technique, specifically the way we administer particles in 2D culture, can influence experimental outcomes. Gold nanoparticles coated with poly(vinylpyrrolidone) were added to J774A.1 mouse monocyte/macrophage cultures as either a concentrated bolus, a bolus then mixed via aspiration, or pre-mixed in cell culture media. Particle-cell interaction was monitored via inductively coupled plasma-optical emission spectroscopy and we found that particles administered in a concentrated dose interacted more with cells compared to the pre-mixed administration method. Spectroscopy studies reveal that the initial formation of the protein corona upon introduction to cell culture media may be responsible for the differences in particle-cell interaction. Modeling of particle deposition using the in vitro sedimentation, diffusion and dosimetry model helped to clarify what particle phenomena may be occurring at the cellular interface. We found that particle administration method in vitro has an effect on particle-cell interactions (i.e. cellular adsorption and uptake). Initial introduction of particles in to complex biological media has a lasting effect on the formation of the protein corona, which in turn mediates particle-cell interaction. It is of note that a minor detail, the way in which we administer particles in cell culture, can have a significant effect on what we observe regarding particle interactions in vitro.
Highlights
The increasing use of engineered nanomaterials in consumer products, development as diagnostic tools and therapeutic vectors, and growing awareness regarding nano-sized pollutants and environmental toxicology has led to a significant push in nanoscience research
We favored the use of PVP19–21 over a more common polymeric coating such as poly(ethylene glycol) (PEG) due to the tendency of PEG to minimize particle uptake22,23. 116 nm AuNP-PVP were administered to cells following three different administration paradigms: (1) NPs pre-mixed in complete cell culture media prior to cell exposure, (2) NPs administered as a concentrated dose to cells in media and mixed via pipetting, or (3) NPs administered as a concentrated dose to cells in media without mixing (Fig. 1a)
Nanoparticles are stable in complete cell culture medium
Summary
The increasing use of engineered nanomaterials in consumer products, development as diagnostic tools and therapeutic vectors, and growing awareness regarding nano-sized pollutants and environmental toxicology has led to a significant push in nanoscience research. We utilized polyvinylpyrrolidone (PVP)-coated, 116 nm diameter gold nanoparticles (AuNP) as a model particle system to study the effects of in vitro administration on particle-cell interaction (Figs 1b and S1). The deposition of AuNP-PVP following administration via the three methods showed that the fraction of the initial dose deposited for concentrated, mixed, and pre-mixed after 24 hr was 0.61, 0.63, and 0.30, respectively (Fig. 3a).
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