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Silver nanoparticles (AgNPs) are widely used in commerce, however, the effect of their physicochemical properties on toxicity remains debatable because of the confounding presence of Ag+ ions. Thus, we designed a series of AgNPs that are stable to surface oxidation and Ag+ ion release. AgNPs were coated with a hybrid lipid membrane comprised of L-phosphatidylcholine (PC), sodium oleate (SOA), and a stoichiometric amount of hexanethiol (HT) to produce oxidant-resistant AgNPs, Ag–SOA–PC–HT. The stability of 7-month aged, 20–100 nm Ag–SOA–PC–HT NPs were assessed using UV–Vis, dynamic light scattering (DLS), and inductively coupled plasma mass spectrometry (ICP-MS), while the toxicity of the nanomaterials was assessed using a well-established, 5-day embryonic zebrafish assay at concentrations ranging from 0–12 mg/L. There was no change in the size of the AgNPs from freshly made samples or 7-month aged samples and minimal Ag+ ion release (<0.2%) in fishwater (FW) up to seven days. Toxicity studies revealed AgNP size- and concentration-dependent effects. Increased mortality and sublethal morphological abnormalities were observed at higher concentrations with smaller nanoparticle sizes. This study, for the first time, determined the effect of AgNP size on toxicity in the absence of Ag+ ions as a confounding variable. 

The controlled synthesis of stable silver nanoparticles (AgNPs), that do not undergo surface oxidation and Ag + ion dissolution, continues to be a major challenge. Here the synthesis of robust hybrid lipid-coated AgNPs, comprised of l -α-phosphatidylcholine (PC) membranes anchored by a stoichiometric amount of long-chained hydrophobic thiols and sodium oleate (SOA) as hydrophobic binding partners, that do not undergo surface oxidation and Ag + ion dissolution, is described. UV-Visible (UV-Vis) spectroscopy, transmission electron microscopy (TEM), and inductively coupled plasma mass spectrometry (ICP-MS) demonstrate that in the presence of strong oxidants, such as potassium cyanide (KCN), the hybrid lipid-coated AgNPs are stable and do not undergo surface oxidation even in the presence of membrane destabilizing surfactants. UV-Vis studies show that the stability of hybrid lipid-coated AgNPs of various sizes and shapes is dependent on the length of the thiol hydrocarbon chain and can be ranked in the order of increasing stability as follows: propanethiol (PT) < hexanethiol (HT) ≤ decanethiol (DT). UV-Vis and ICP-MS studies show that the hybrid lipid-coated AgNPs do not change in size or shape confirming that the AgNPs do not undergo surface oxidation and Ag + ion dissolution when placed in the presence of strong oxidants, chlorides, thiols, and low pH. Long-term stability studies, over 21 days, show that the hybrid lipid-coated AgNPs do not release Ag + ions and are more stable. Overall, these studies demonstrate hybrid membrane encapsulation of nanomaterials is a viable method for stabilizing AgNPs in a “shape-locked” form that is unable to undergo surface oxidation, Ag + ion release, aging, or shape conversion. More importantly, this design strategy is a simple approach to the synthesis and stabilization of AgNPs for a variety of biomedical and commercial applications where Ag + ion release and toxicity is a concern. With robust and shielded AgNPs, investigators can now evaluate and correlate how the physical features of AgNPs influence toxicity without the confounding factor of Ag + ions present in samples. This design strategy also provides an opportunity where the membrane composition can be tuned to control the release rate of Ag + ions for optimizing antimicrobial activity.