Neuronal haemoglobin induces loss of dopaminergic neurons in mouse Substantia nigra, cognitive deficits and cleavage of endogenous α-synuclein.

Parkinson's disease (PD) presents the selective loss of A9 dopaminergic (DA) neurons of Substantia Nigra pars compacta (SNpc) and the presence of intracellular aggregates called Lewy bodies. α-synuclein (α-syn) species truncated at the carboxy-terminal (C-terminal) accumulate in pathological inclusions and promote α-syn aggregation and toxicity. Haemoglobin (Hb) is the major oxygen carrier protein in erythrocytes. In addition, Hb is expressed in A9 DA neurons where it influences mitochondrial activity. Hb overexpression increases cells' vulnerability in a neurochemical model of PD in vitro and forms cytoplasmic and nucleolar aggregates upon short-term overexpression in mouse SNpc. In this study, α and ß-globin chains were co-expressed in DA cells of SNpc in vivo upon stereotaxic injections of an Adeno-Associated Virus isotype 9 (AAV9) and in DA iMN9D cells in vitro. Long-term Hb over-expression in SNpc induced the loss of about 50% of DA neurons, mild motor impairments, and deficits in recognition and spatial working memory. Hb triggered the formation of endogenous α-syn C-terminal truncated species. Similar α-syn fragments were found in vitro in DA iMN9D cells over-expressing α and ß- globins when treated with pre-formed α-syn fibrils. Our study positions Hb as a relevant player in PD pathogenesis for its ability to trigger DA cells' loss in vivo and the formation of C-terminal α-syn fragments.

Carbon Nanotubes Substrates Alleviate Pro-Calcific Evolution in Porcine Valve Interstitial Cells.

The quest for surfaces able to interface cells and modulate their functionality has raised, in recent years, the development of biomaterials endowed with nanocues capable of mimicking the natural extracellular matrix (ECM), especially for tissue regeneration purposes. In this context, carbon nanotubes (CNTs) are optimal candidates, showing dimensions and a morphology comparable to fibril ECM constituents. Moreover, when immobilized onto surfaces, they demonstrated outstanding cytocompatibility and ease of chemical modification with ad hoc functionalities. In this study, we interface porcine aortic valve interstitial cells (pVICs) to multi-walled carbon nanotube (MWNT) carpets, investigating the impact of surface nano-morphology on cell properties. The results obtained indicate that CNTs significantly affect cell behavior in terms of cell morphology, cytoskeleton organization, and mechanical properties. We discovered that CNT carpets appear to maintain interfaced pVICs in a sort of "quiescent state", hampering cell activation into a myofibroblasts-like phenotype morphology, a cellular evolution prodromal to Calcific Aortic Valve Disease (CAVD) and characterized by valve interstitial tissue stiffening. We found that this phenomenon is linked to CNTs' ability to alter cell tensional homeostasis, interacting with cell plasma membranes, stabilizing focal adhesions and enabling a better strain distribution within cells. Our discovery contributes to shedding new light on the ECM contribution in modulating cell behavior and will open the door to new criteria for designing nanostructured scaffolds to drive cell functionality for tissue engineering applications.

Correction: Structural insights into fusion mechanisms of small extracellular vesicles with model plasma membranes.

Correction for 'Structural insights into fusion mechanisms of small extracellular vesicles with model plasma membranes' by Fabio Perissinotto et al., Nanoscale, 2021, 13, 5224-5233, DOI: .

Structural insights into fusion mechanisms of small extracellular vesicles with model plasma membranes.

Extracellular vesicles (EVs) are a potent intercellular communication system. Such small vesicles transport biomolecules between cells and throughout the body, strongly influencing the fate of recipient cells. Due to their specific biological functions they have been proposed as biomarkers for various diseases and as optimal candidates for therapeutic applications. Despite their extreme biological relevance, their mechanisms of interaction with the membranes of recipient cells are still hotly debated. Here, we propose a multiscale investigation based on atomic force microscopy, small angle X-ray scattering, small angle neutron scattering and neutron reflectometry to reveal structure-function correlations of purified EVs in interaction with model membrane systems of variable complex compositions and to spot the role of different membrane phases on the vesicle internalization routes. Our analysis reveals strong interactions of EVs with the model membranes and preferentially with the borders of protruding phase domains. Moreover, we found that upon vesicle breaking on the model membrane surface, the biomolecules carried by/on EVs diffuse with different kinetics rates, in a process distinct from simple fusion. The biophysical platform proposed here has clear implications on the modulation of EV internalization routes by targeting specific domains at the plasma cell membrane and, as a consequence, on EV-based therapies.

Iron-mediated interaction of alpha synuclein with lipid raft model membranes.

The aberrant misfolding and aggregation of alpha synuclein (αS) into toxic oligomeric species is one of the key features associated with the pathogenesis of Parkinson's disease (PD). It involves different biochemical and biophysical factors as plasma membrane binding and interaction with heavy metal ions. In the present work, atomic force microscopy (AFM) is combined with Fourier Transform Infrared Spectroscopy (FTIR) measurements to investigate the interaction of wild-type (WT) and A53T mutated alpha synuclein with artificial lipid bilayers mimicking lipid raft (LR) domains, before and after ferrous cations (Fe2+) treatment. In the absence of iron, protein monomers produce a thinning of the membrane, targeting the non-raft phase of the bilayer preferentially. On the contrary, iron actively promotes the formation of globular protein aggregates, resembling oligomers, targeted to LR domains. In both aggregation states, monomer and oligomer, the mutated A53T protein exhibits a greater and faster membrane-interaction. These results underlie a new mechanism of membrane-protein interaction in PD. The targeting of Fe2+-promoted αS oligomers to LRs might be functional for the disease and be helpful for the development of new therapeutic strategies.

Do Chlorhexidine and Probiotics Solutions Provoke Corrosion of Orthodontic Mini-implants? An In Vitro Study.

PURPOSE: The aim of this study was to explore the surface roughness and hardness of the implant head of orthodontic mini-implants made from different alloys before and after their in vitro exposure to agents for prevention of gingivitis, mucositis, and peri-implantitis: chlorhexidine and probiotics. MATERIALS AND METHODS: Three types of commercially available mini-implants were tested: 316 stainless steel, titanium Grade 5, and titanium Grade 23 (both Ti-6Al-4V alloys with the same atomic weight percentage of Ti, Al, and V, with the difference being in maximal reduction of O2 in Grade 23 to 0.13% of atomic weight). They were immersed in three experimental solutions: artificial saliva, saliva with probiotic bacteria Lactobacillus reuteri, and saliva with oral antiseptic chlorhexidine (CHX). Samples were immersed for 28 days, thermocycled, then stored in an incubator at 37°C. Surface roughness and microhardness on five samples of each of the three implant types were measured by atomic force microscopy and the Vickers method, respectively. RESULTS: Exposure of titanium implant Grade 5 to probiotics significantly increased roughness compared with other media (P < .005). Exposure to CHX significantly increased the roughness of steel implants (P < .05). Neither saliva, probiotic, nor CHX altered microhardness of titanium implants significantly. In steel implants, the exposure to CHX and probiotics decreased microhardness compared with unexposed implants (P < .031), but not in comparison to saliva. CONCLUSION: Probiotics seem to increase roughness of titanium mini-implants, while CHX seems to increase roughness of steel mini-implants. Only stainless steel implants had an altered, decreased hardness after exposure to CHX, although the same was found after their exposure to saliva. For patients undergoing orthodontic treatment with temporary anchorage units, CHX could be recommended for titanium, and probiotics for stainless steel mini-implants in oral-hygiene maintenance.

Mixed Fluorinated/Hydrogenated Self-Assembled Monolayer-Protected Gold Nanoparticles: In Silico and In Vitro Behavior.

Gold nanoparticles (AuNPs) covered with mixtures of immiscible ligands present potentially anisotropic surfaces that can modulate their interactions at complex nano-bio interfaces. Mixed, self-assembled, monolayer (SAM)-protected AuNPs, prepared with incompatible hydrocarbon and fluorocarbon amphiphilic ligands, are used here to probe the molecular basis of surface phase separation and disclose the role of fluorinated ligands on the interaction with lipid model membranes and cells, by integrating in silico and experimental approaches. These results indicate that the presence of fluorinated amphiphilic ligands enhances the membrane binding ability and cellular uptake of gold nanoparticles with respect to those coated only with hydrogenated amphiphilic ligands. For mixed monolayers, computational results suggest that ligand phase separation occurs on the gold surface, and the resulting anisotropy affects the number of contacts and adhesion energies with a membrane bilayer. This reflects in a diverse membrane interaction for NPs with different surface morphologies, as determined by surface plasmon resonance, as well as differential effects on cells, as observed by flow cytometry and confocal microscopy. Overall, limited changes in monolayer features can significantly affect NP surface interfacial properties, which, in turn, affect the interaction of SAM-AuNPs with cellular membranes and subsequent effects on cells.

Vitamin C/Stearic Acid Hybrid Monolayer Adsorption at Air-Water and Air-Solid Interfaces.

Because of the antioxidant activity of vitamin C (Vit C) polar heads, they can be used as a protective agent for fatty acids. Hence, the study on the growth of Vit C/stearic acid (SA) mixed binary films at air-water interface (known as Langmuir monolayer) and air-solid interface (known as Langmuir-Blodgett films) is of paramount interest. Although Vit C is situated at subsurface beneath SA molecules and interacts via hydrogen bonding between the hydroxyl groups of Vit C and SA, several Vit C molecules may infiltrate within SA two-dimensional matrix at the air-water interface. The increased mole fraction of Vit C (0.125-0.5) and the reduction of temperature (from 22 to 10 °C) of the subphase water result in an increase in the amount of adsorbed Vit C at the air-water interface. The surface pressure (π)-area (A) isotherms illustrate that such inclusion of Vit C provokes a spreading out of Vit C/SA binary monolayers, which leads to an alteration of different physicochemical parameters such as elasticity, Gibbs free energy of mixing, enthalpy, entropy, interaction energy parameter, and activity coefficient. However, being polar in nature, the transfer of pure Vit C on substrates gets affected. It can be transferred onto substrate by mixing suitably with SA as confirmed by infrared spectra. Their structures, extracted X-ray reflectivity, and atomic force microscopy (topography and phase imaging) are found to be strongly dependent on the nature of the substrate (hydrophilic and hydrophobic).

Sculpting neurotransmission during synaptic development by 2D nanostructured interfaces.

Carbon nanotube-based biomaterials critically contribute to the design of many prosthetic devices, with a particular impact in the development of bioelectronics components for novel neural interfaces. These nanomaterials combine excellent physical and chemical properties with peculiar nanostructured topography, thought to be crucial to their integration with neural tissue as long-term implants. The junction between carbon nanotubes and neural tissue can be particularly worthy of scientific attention and has been reported to significantly impact synapse construction in cultured neuronal networks. In this framework, the interaction of 2D carbon nanotube platforms with biological membranes is of paramount importance. Here we study carbon nanotube ability to interfere with lipid membrane structure and dynamics in cultured hippocampal neurons. While excluding that carbon nanotubes alter the homeostasis of neuronal membrane lipids, in particular cholesterol, we document in aged cultures an unprecedented functional integration between carbon nanotubes and the physiological maturation of the synaptic circuits.