Hybrid Amyloid-Chitin Nanofibrils for Magnetic and Catalytic Aerogels.

In the quest for a sustainable and circular economy, it is essential to explore environmentally friendly alternatives to traditional petroleum-based materials. A promising pathway toward this goal lies in the leveraging of biopolymers derived from food waste, such as proteins and polysaccharides, to develop advanced sustainable materials. Here, we design versatile hybrid materials by hybridizing amyloid nanofibrils derived by self-assembly of whey, a dairy byproduct, with chitin nanofibrils exfoliated from the two distinct allomorphs of α-chitin and ß-chitin, extracted from seafood waste. Various hydrogels and aerogels were developed via the hybridization and reassembly of these biopolymeric nanobuilding blocks, and they were further magnetized upon biomineralization with iron nanoparticles. The pH-phase diagram highlights the significant role of electrostatic interactions in gel formation, between positively charged amyloid fibrils and negatively charged chitin nanofibrils. Hybrid magnetic aerogels exhibit a ferromagnetic response characterized by a low coercivity (<50 Oe) and a high specific magnetization (>40 emu/g) at all temperatures, making them particularly suitable for superparamagnetic applications. Additionally, these aerogels exhibit a distinct magnetic transition, featuring a higher blocking temperature (200 K) compared to previously reported similar nanoparticles (160 K), indicating enhanced magnetic stability at elevated temperatures. Finally, we demonstrate the practical application of these hybrid magnetic materials as catalysts for carbon monoxide oxidation, showcasing their potential in environmental pollution control and highlighting their versatility as catalyst supports.

Multi-length scale structural investigation of lysozyme self-assembly.

Reactive amyloid oligomers are responsible for cytotoxicity in amyloid pathologies and because of their unstable nature characterizing their behavior is a challenge. The physics governing the self-assembly of proteins in crowded conditions is extremely complex and its comprehension, despite its paramount relevance to understanding molecular mechanisms inside cells and optimizing pharmaceutical processes, remains inconclusive. Here, we focus on the amyloid oligomerization process in self-crowded lysozyme aqueous solutions in acidic conditions. We reveal that the amyloid oligomers form at high protein concentration and low pH. Through multi-length scale spectroscopic investigations, we find that amyloid oligomers can further interconnect with each other by weak and non-specific interactions forming an extended network that leads to the percolation of the whole system. Our multi-length scale structural analysis follows the thermal history of amyloid oligomers from different perspectives and highlights the impact of hierarchical self-assembly of biological macromolecules on functional properties.

Unraveling gelation kinetics, arrested dynamics and relaxation phenomena in filamentous colloids by photon correlation imaging.

The fundamental understanding of the gelation kinetics, stress relaxation and temporal evolution in colloidal filamentous gels is central to many aspects of soft and biological matter, yet a complete description of the inherent complex dynamics of these systems is still missing. By means of photon correlation imaging (PCI), we studied the gelation of amyloid fibril solutions, chosen as a model filamentous colloid with immediate significance to biology and nanotechnology, upon passage of ions through a semi-permeable membrane. We observed a linear-in-time evolution of the gelation front and rich rearrangement dynamics of the gels, the magnitude and the spatial propagation of which depend on how effectively electrostatic interactions are screened by different ionic strengths. Our analysis confirms the pivotal role of salt concentration in tuning the properties of amyloid gels, and suggests potential routes for explaining the physical mechanisms behind the linear advance of the salt ions.

Oat Plant Amyloids for Sustainable Functional Materials.

Amyloid functional materials from amyloid fibril building blocks, produced in vitro from amyloidogenic natural proteins or synthetic peptides, show diverse functionalities ranging from environmental science and biomedicine, to nanotechnology and biomaterials. However, sustainable and affordable sources of amyloidogenic proteins remain the bottleneck for large-scale applications, and to date, interest remains essentially limited to fundamental studies. Plant-derived proteins would be an ideal source due to their natural abundance and low environmental impact. Hereby oat globulin, the primary protein of oat plant (Avena sativa), is utilized to yield high-quality amyloid fibrils and functional materials based thereof. These fibrils show a rich multistranded ribbon-like polymorphism and a fibrillization process with both irreversible and reversible pathways. The authors furthermore fabricate oat-amyloid aerogels, films, and membranes for possible use in water purification, sensors, and patterned electrodes. The sustainability footprint of oat-amyloids against other protein sources is demonstrated, anticipating an environmentally-efficient platform for advanced materials and technologies.

Polysaccharide-reinforced amyloid fibril hydrogels and aerogels.

ß-Lactoglobulin amyloid fibrils are bio-colloids of high interest in many fields (e.g. water purification, cell growth, drug delivery and sensing). While the mechanical properties of pure amyloid fibril gels meet the needs of some applications, mechanical fragility often hinders a wider usage basin. In this work, we present a simple and sustainable approach for reinforcing amyloid fibril hydrogels and aerogels, upon the diffusion of polysaccharides (low-acetylated Gellan Gum and κ-carrageenan) inside their mesh. The formed hybrid materials show enhanced resistance upon compression, without any loss of the exquisite surface reactivity of the amyloid fibrils. The proposed approach can pave the way for designing composite materials that are both highly functional and environmentally friendly.

Sustainable Removal of Microplastics and Natural Organic Matter from Water by Coagulation-Flocculation with Protein Amyloid Fibrils.

Water contamination is a global threat due to its damaging effects on the environment and human health. Water pollution by microplastics (MPs), dissolved natural organic matter (NOM), and other turbid particles is ubiquitous in water treatment. Here, we introduce lysozyme amyloid fibrils as a novel natural bio-flocculant and explore their ability to flocculate and precipitate the abovementioned undesired colloidal objects. Thanks to their positively charged surface in a very broad range of pH, lysozyme amyloid fibrils show an excellent turbidity removal efficiency of 98.2 and 97.9% for dispersed polystyrene MPs and humic acid (HA), respectively. Additionally, total organic carbon measurements confirm these results by exhibiting removal efficiencies of 93.4 and 61.9% for purifying water from dispersed MPs and dissolved HA, respectively. The comparison among amyloid fibrils, commercial flocculants (FeCl3 and polyaluminumchloride), and native lysozyme monomers points to the superiority of amyloid fibrils at the same dosage and sedimentation time. Furthermore, the turbidity of pristine and MP-spiked wastewater and lake water decreased after the treatment by amyloid fibrils, validating their coagulation-flocculation performance under natural conditions. All these results demonstrate lysozyme amyloid fibrils as an appropriate natural bio-flocculant for removing dispersed MPs, NOM, and turbid particles from water.

VEGF and VEGFR2 bind to similar pH-sensitive sites on fibronectin, exposed by heparin-mediated conformational changes.

Physical interactions between vascular endothelial growth factor (VEGF), a central player in blood endothelial cell biology, and fibronectin, a major fibrillar protein of the extracellular matrix, are important determinants of angiogenic activity in health and disease. Conditions signaling the need for new blood vessel growth, such as hypoxia and low extracellular pH, increase VEGF-fibronectin interactions. These interactions can be further fine-tuned through changes in the availability of the VEGF-binding sites on fibronectin, regulated by conformational changes induced by heparin and heparan sulfate chains within the extracellular matrix. These interactions may alter VEGF bioavailability, generate gradients, or alter the way VEGF is recognized by and activates its cell-surface receptors. Here, using equilibrium and kinetic studies, we discovered that fibronectin can also interact with the extracellular domain of the VEGF receptor 2 (VEGFR2). The VEGFR2-binding sites on fibronectin show great similarity to the VEGF-binding sites, as they were also exposed upon heparin-induced conformational changes in fibronectin, and the interaction was enhanced at acidic pH. Kinetic parameters and affinities for VEGF and VEGFR2 binding to fibronectin were determined by surface plasmon resonance measurements, revealing two populations of fibronectin-binding sites for each molecule. Our data also suggest that a VEGF/VEGFR2/fibronectin triple complex may be formed by VEGF or VEGFR2 first binding to fibronectin and subsequently recruiting the third binding partner. The formation of such a complex may lead to the activation of distinct angiogenic signaling pathways, offering new possibilities for clinical applications that target angiogenesis.

Investigating the Mechanism of Cyclodextrins in the Treatment of Niemann-Pick Disease Type C Using Crosslinked 2-Hydroxypropyl-ß-cyclodextrin.

Niemann-Pick disease type C (NPC) is a severe disorder that is characterized by intracellular transport abnormalities leading to cytoplasmic accumulation of lipids such as cholesterol and sphingolipids. The compound 2-hydroxypropyl-ß-cyclodextrin (HPßCD) has high cholesterol complexation capacity and is currently under clinical investigation for the NPC treatment. However, due to its short blood half-life, high doses are required to produce a therapeutic effect. In this work, stable polymerized HPßCD is generated to investigate their in vitro mechanisms of action and in vivo effects. Crosslinked CDs (8-312 kDa) display a ninefold greater cholesterol complexation capacity than monomeric HPßCD but are taken up to a lower extent, resulting in an overall comparable in vitro effect. In vivo, the 19.3 kDa HPßCD exhibits a longer half-life than the monomeric HPßCD but it does not increase the life span of Npc1 mice, possibly due to reduced brain penetration. This is circumvented by the application of magnetic resonance imaging-guided low intensity-pulsed focused ultrasound (MRIg-FUS), which increases the brain penetration of the CD. In conclusion, stable polymerized HPßCDs can elucidate CDs' mechanism of action while the use of MRIg-FUS warrants further investigation, as it may be key to harnessing CDs full therapeutic potential in the NPC treatment.