Inhibition of Amyloid Aggregation and Toxicity with Janus Iron Oxide Nanoparticles.

Amyloid aggregation is a ubiquitous form of protein misfolding underlying the pathologies of Alzheimer's disease (AD), Parkinson's disease (PD) and type 2 diabetes (T2D), three primary forms of human amyloid diseases. While much has been learned about the origin, diagnosis and management of these neurological and metabolic disorders, no cure is currently available due in part to the dynamic and heterogeneous nature of the toxic oligomers induced by amyloid aggregation. Here we synthesized beta casein-coated iron oxide nanoparticles (ßCas IONPs) via a BPA-P(OEGA-b-DBM) block copolymer linker. Using a thioflavin T kinetic assay, transmission electron microscopy, Fourier transform infrared spectroscopy, discrete molecular dynamics simulations and cell viability assays, we examined the Janus characteristics and the inhibition potential of ßCas IONPs against the aggregation of amyloid beta (Aß), alpha synuclein (αS) and human islet amyloid polypeptide (IAPP) which are implicated in the pathologies of AD, PD and T2D. Incubation of zebrafish embryos with the amyloid proteins largely inhibited hatching and elicited reactive oxygen species, which were effectively rescued by the inhibitor. Furthermore, Aß-induced damage to mouse brain was mitigated in vivo with the inhibitor. This study revealed the potential of Janus nanoparticles as a new nanomedicine against a diverse range of amyloid diseases.

Dynamic Protein Corona of Gold Nanoparticles with an Evolving Morphology.

Much has been learned about the protein coronae and their biological implications within the context of nanomedicine and nanotoxicology. However, no data is available about the protein coronae associated with nanoparticles undergoing spontaneous surface-energy minimization, a common phenomenon during the synthesis and shelf life of nanomaterials. Accordingly, here we employed gold nanoparticles (AuNPs) possessing the three initial states of spiky, midspiky, and spherical shapes and determined their acquisition of human plasma protein coronae with label-free mass spectrometry. The AuNPs collected coronal proteins that were different in abundance, physicochemical parameters, and interactive biological network. The size and structure of the coronal proteins matched the morphology of the AuNPs, where small globular proteins and large fibrillar proteins were enriched on spiky AuNPs, while large proteins were abundant on spherical AuNPs. Furthermore, the AuNPs induced endothelial leakiness to different degrees, which was partially negated by their protein coronae as revealed by confocal fluorescence microscopy, in vitro and ex vivo transwell assays, and signaling pathway assays. This study has filled a knowledge void concerning the dynamic protein corona of nanoparticles possessing an evolving morphology and shed light on their implication for future nanomedicine harnessing the paracellular pathway.

A Framework of Paracellular Transport via Nanoparticles-Induced Endothelial Leakiness.

Nanomaterial-induced endothelial leakiness (NanoEL) is an interfacial phenomenon denoting the paracellular transport of nanoparticles that is pertinent to nanotoxicology, nanomedicine and biomedical engineering. While the NanoEL phenomenon is complementary to the enhanced permeability and retention effect in terms of their common applicability to delineating the permeability and behavior of nanoparticles in tumoral environments, these two effects significantly differ in scope, origin, and manifestation. In the current study, the descriptors are fully examined of the NanoEL phenomenon elicited by generic citrate-coated gold nanoparticles (AuNPs) of changing size and concentration, from microscopic gap formation and actin reorganization down to molecular signaling pathways and nanoscale interactions of AuNPs with VE-cadherin and its intra/extracellular cofactors. Employing synergistic in silico methodologies, for the first time the molecular and statistical mechanics of cadherin pair disruption, especially in response to AuNPs of the smallest size and highest concentration are revealed. This study marks a major advancement toward establishing a comprehensive NanoEL framework for complementing the understanding of the transcytotic pathway and for guiding the design and application of future nanomedicines harnessing the myriad functions of the mammalian vasculature.

Spontaneous Formation of ß-sheet Nano-barrels during the Early Aggregation of Alzheimer's Amyloid Beta.

Soluble low-molecular-weight oligomers formed during the early aggregation of amyloid peptides have been hypothesized as a major toxic species of amyloidogenesis. Herein, we performed the first synergic in silico, in vitro and in vivo validations of the structure, dynamics and toxicity of Aß42 oligomers. Aß peptides readily assembled into ß-rich oligomers comprised of extended ß-hairpins and ß-strands. Nanosized ß-barrels were observed with certainty with simulations, transmission electron microscopy and Fourier transform infrared spectroscopy, corroborated by immunohistochemistry, cell viability, apoptosis, inflammation, autophagy and animal behavior assays. Secondary and tertiary structural proprieties of these oligomers, such as the sequence regions with high ß-sheet propensities and inter-residue contact frequency patterns, were similar to the properties known for Aß fibrils. The unambiguous spontaneous formation of ß-barrels in the early aggregation of Aß42 supports their roles as the common toxic intermediates in Alzheimer's pathobiology and a target for Alzheimer's therapeutics.

A tandem activation of NLRP3 inflammasome induced by copper oxide nanoparticles and dissolved copper ion in J774A.1 macrophage.

For the first time, we reported that CuONPs exposure induced interleukin (IL)-1ß-mediated inflammation via NOD-, LRR- and pyrin domain-containing protein 3 (NLRP3) inflammasome in J774A.1 macrophage. Mechanistically, CuONPs activated NLRP3 inflammasome is a two-fold process. Firstly, CuONPs challenge caused lysosomal damage, along with the release of cathepsin B, which directly mediated the activation of NLRP3 inflammasomes. Interestingly, after the deposition in lysosomes, CuONPs may release copper ion due to the acidic environment of lysosomes. Consequently, the released copper ions significantly induced cellular oxidative stress and further mediated the activation of NLRP3 inflammasomes. Moreover, CuONPs exposure could prime J774A.1 macrophage to express pro-IL-1ß through myeloid differentiation factor 88 (MyD88)-dependent Toll-like receptor 4 (TLR4) signal pathway subsequently activating nuclear transcription factor kappa B (NF-κB).

Impact of Protein Corona on Noncovalent Molecule-Gold Nanoparticle-Based Sensing.

Gold nanoparticle (AuNP)-based sensors have been extensively applied for sensing or imaging. It is known that a protein shell named protein corona (PC) formed around the nanomaterials could not only block the desired function of nanomaterials but also affect their behavior, which is a hot and important issue needing consideration. Therefore, we hypothesize that the formation of PC around AuNPs could inevitably affect the AuNP-based target assay. In this work, the effects of PC on the detection results in sensors based on AuNPs were studied. Three types of noncovalent molecule-AuNP sensors including AuNP-dichlorofluorescein, AuNP-aptamer, and AuNP-antibody-DNA were constructed, and several typical proteins (bovine serum albumin, fibrinogen, hemoglobin, and ß-lactoglobulin), milk, and fetal bovine serum were selected as models for the formation of PCs. This study shows that the PC could cause the loss of detection signals (up to 80%) and result in positive deviation of the measuring value compared with the true value. Moreover, the loss of detection signals could also increase the limits of detection (almost 10 times), decreasing the sensitivity of the three types of sensors, as proposed in this work compared to that without PC. Moreover, the polyethylene glycol backfilling strategy could not resolve the negative effects of PC on noncovalent molecule-AuNP sensors. The impacts of PC on detection results from noncovalent molecule-AuNP sensors would cause misdiagnosis or wasted production, which needs careful reconsideration of the AuNP-based detection in application fields like clinic diagnosis, food safety control, and so forth.

Determination of 3-Methyl-quinoxaline-2-carboxylic Acid and Quinoxaline-2-carboxylic Acid in Pork Based on a Background Fluorescence Quenching Immunochromatographic Assay.

A novel rapid method based on a background fluorescence quenching immunochromatographic assay (bFQICA) was established to achieve simultaneously the quantitative detection of 3-methyl-quinoxaline-2-carboxylic acid (MQCA) and quinoxaline-2-carboxylic acid (QCA), which were efficiently extracted and enriched 4 times using immunomagnetic beads from pork. The analysis of field pork samples by bFQICA was in accordance with that of LC-MS/MS; especially, the proposed bFQICA exhibited great advantages in convenience and efficiency, which only takes 30 min for the detection of MQCA and QCA.