New approach for time-resolved and dynamic investigations on nanoparticles agglomeration

Nanoparticle (NP) colloidal stability plays a crucial role in biomedical application not only for human and environmental safety but also for NP efficiency and functionality. NP agglomeration is considered as a possible process in monodispersed NP colloidal solutions, which drastically affects colloidal stability. This process is triggered by changes in the physicochemical properties of the surrounding media, such as ionic strength (IS), pH value, or presence of biomolecules. Despite different available characterization methods for nanoparticles (NPs), there is a lack of information about the underlying mechanisms at the early stage of dynamic behaviors, namely changing in NP size distribution and structure while placing them from a stable colloidal solution to a new media like biological fluids. In this study, an advanced in situ approach is presented that combines small angle X-ray scattering (SAXS) and microfluidics, allowing label-free, direct, time-resolved, and dynamic observations of the early stage of NP interaction/agglomeration initiated by environmental changes. It is shown for silica NPs that the presence of protein in the media enormously accelerates the NP agglomeration process compared to respective changes in IS and pH. High IS results in a staring agglomeration process after 40 min, though, in case of protein presence in media, this time decreased enormously to 48 s. These time scales show that this method is sensitive and precise in depicting the dynamics of fast and slow NP interactions in colloidal conditions and therefore supports understanding the colloidal stability of NPs in various media concluding in safe and efficient NP designing for various applications

Nanoscale studies link amyloid maturity with polyglutamine diseases onset

The presence of expanded poly-glutamine (polyQ) repeats in proteins is directly linked to the pathogenesis of several neurodegenerative diseases, including Huntington's disease. However, the molecular and structural basis underlying the increased toxicity of aggregates formed by proteins containing expanded polyQ repeats remain poorly understood, in part due to the size and morphological heterogeneity of the aggregates they form in vitro. To address this knowledge gap and technical limitations, we investigated the structural, mechanical and morphological properties of fibrillar aggregates at the single molecule and nanometer scale using the first exon of the Huntingtin protein as a model system (Exon1). Our findings demonstrate a direct correlation of the morphological and mechanical properties of Exon1 aggregates with their structural organization at the single aggregate and nanometric scale and provide novel insights into the molecular and structural basis of Huntingtin Exon1 aggregation and toxicity

Unraveling the complexity of amyloid polymorphism using gold nanoparticles and cryo-EM

Increasing evidence suggests that amyloid polymorphism gives rise to different strains of amyloids with distinct toxicities and pathology-spreading properties. Validating this hypothesis is challenging due to a lack of tools and methods that allow for the direct characterization of amyloid polymorphism in hydrated and complex biological samples. Here, we report on the development of 11-mercapto-1-undecanesulfonate-coated gold nanoparticles (NPs) that efficiently label the edges of synthetic, recombinant, and native amyloid fibrils derived from different amyloidogenic proteins. We demonstrate that these NPs represent powerful tools for assessing amyloid morphological polymorphism, using cryogenic transmission electron microscopy (cryo-EM). The NPs allowed for the visualization of morphological features that are not directly observed using standard imaging techniques, including transmission electron microscopy with use of the negative stain or cryo-EM imaging. The use of these NPs to label native paired helical filaments (PHFs) from the postmortem brain of a patient with Alzheimer's disease, as well as amyloid fibrils extracted from the heart tissue of a patient suffering from systemic amyloid light-chain amyloidosis, revealed a high degree of homogeneity across the fibrils derived from human tissue in comparison with fibrils aggregated in vitro. These findings are consistent with, and strongly support, the emerging view that the physiologic milieu is a key determinant of amyloid fibril strains. Together, these advances should not only facilitate the profiling and characterization of amyloids for structural studies by cryo-EM, but also pave the way to elucidate the structural basis of amyloid strains and toxicity, and possibly the correlation between the pathological and clinical heterogeneity of amyloid diseases