Interfacing Graphene-Based Materials With Neural Cells

The scientific community has witnessed an exponential increase in the applications of graphene and graphene-based materials in a wide range of fields, from engineering to electronics to biotechnologies and biomedical applications. For what concerns neuroscience, the interest raised by these materials is two-fold. On one side, nanosheets made of graphene or graphene derivatives (graphene oxide, or its reduced form) can be used as carriers for drug delivery. Here, an important aspect is to evaluate their toxicity, which strongly depends on flake composition, chemical functionalization and dimensions. On the other side, graphene can be exploited as a substrate for tissue engineering. In this case, conductivity is probably the most relevant amongst the various properties of the different graphene materials, as it may allow to instruct and interrogate neural networks, as well as to drive neural growth and differentiation, which holds a great potential in regenerative medicine. In this review, we try to give a comprehensive view of the accomplishments and new challenges of the field, as well as which in our view are the most exciting directions to take in the immediate future. These include the need to engineer multifunctional nanoparticles (NPs) able to cross the blood-brain-barrier to reach neural cells, and to achieve on-demand delivery of specific drugs. We describe the state-of-the-art in the use of graphene materials to engineer three-dimensional scaffolds to drive neuronal growth and regeneration in vivo, and the possibility of using graphene as a component of hybrid composites/multi-layer organic electronics devices. Last but not least, we address the need of an accurate theoretical modeling of the interface between graphene and biological material, by modeling the interaction of graphene with proteins and cell membranes at the nanoscale, and describing the physical mechanism(s) of charge transfer by which the various graphene materials can influence the excitability and physiology of neural cells

Interactions between Primary Neurons and Graphene Films with Different Structure and Electrical Conductivity

Graphene-based materials represent a useful tool for the realization of novel neural interfaces. Several studies have demonstrated the biocompatibility of graphene-based supports, but the biological interactions between graphene and neurons still pose open questions. In this work, the influence of graphene films with different characteristics on the growth and maturation of primary cortical neurons is investigated. Graphene films are grown by chemical vapor deposition progressively lowering the temperature range from 1070 to 650 °C to change the lattice structure and corresponding electrical conductivity. Two graphene-based films with different electrical properties are selected and used as substrate for growing primary cortical neurons: i) highly crystalline and conductive (grown at 1070 °C) and ii) highly disordered and 140-times less conductive (grown at 790 °C). Electron and fluorescence microscopy imaging reveal an excellent neuronal viability and the development of a mature, structured, and excitable network onto both substrates, regardless of their microstructure and electrical conductivity. The results underline that high electrical conductivity by itself is not fundamental for graphene-based neuronal interfaces, while other physico–chemical characteristics, including the atomic structure, should be also considered in the design of functional, bio-friendly templates. This finding widens the spectrum of carbon-based materials suitable for neuroscience applications

Membrane Environment Enables Ultrafast Isomerization of Amphiphilic Azobenzene

G.M.P. and E.C. contributed equally to this work. G.M.P. acknowledges the financial support from Fondazione Cariplo, grant no. 2018-0979. The authors thank the financial support from the EU Horizon 2020 Research and Innovation Programme under Grant Agreement No. 643238 (SYNCHRONICS). The authors also thank Dr. Daniele Viola for helping with the analysis of the TA data.The non‐covalent affinity of photoresponsive molecules to biotargets represents an attractive tool for achieving effective cell photo‐stimulation. Here, an amphiphilic azobenzene that preferentially dwells within the plasma membrane is studied. In particular, its isomerization dynamics in different media is investigated. It is found that in molecular aggregates formed in water, the isomerization reaction is hindered, while radiative deactivation is favored. However, once protected by a lipid shell, the photochromic molecule reacquires its ultrafast photoisomerization capacity. This behavior is explained considering collective excited states that may form in aggregates, locking the conformational dynamics and redistributing the oscillator strength. By applying the pump probe technique in different media, an isomerization time in the order of 10 ps is identified and the deactivation in the aggregate in water is also characterized. Finally, it is demonstrated that the reversible modulation of membrane potential of HEK293 cells via illumination with visible light can be indeed related to the recovered trans→cis photoreaction in lipid membrane. These data fully account for the recently reported experiments in neurons, showing that the amphiphilic azobenzenes, once partitioned in the cell membrane, are effective light actuators for the modification of the electrical state of the membrane.Fondazione Cariplo. Grant Number: 2018‐0979EU Horizon 2020 Research and Innovation Programme. Grant Number: 64323

APache Is an AP2-Interacting Protein Involved in Synaptic Vesicle Trafficking and Neuronal Development

Synaptic transmission is critically dependent on synaptic vesicle (SV) recycling. Although the precise mechanisms of SV retrieval are still debated, it is widely accepted that a fundamental role is played by clathrin-mediated endocytosis, a form of endocytosis that capitalizes on the clathrin/adaptor protein complex 2 (AP2) coat and several accessory factors. Here, we show that the previously uncharacterized protein KIAA1107, predicted by bioinformatics analysis to be involved in the SV cycle, is an AP2-interacting clathrin-endocytosis protein (APache). We found that APache is highly enriched in the CNS and is associated with clathrin-coated vesicles via interaction with AP2. APache-silenced neurons exhibit a severe impairment of maturation at early developmental stages, reduced SV density, enlarged endosome-like structures, and defects in synaptic transmission, consistent with an impaired clathrin/AP2-mediated SV recycling. Our data implicate APache as an actor in the complex regulation of SV trafficking, neuronal development, and synaptic plasticity

Chronic Stress Induces Sex-Specific Alterations in Methylation and Expression of Corticotropin-Releasing Factor Gene in the Rat

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Interactions between Primary Neurons and Graphene Films with Different Structure and Electrical Conductivity

Graphene-based materials represent a useful tool for the realization of novel neural interfaces. Several studies have demonstrated the biocompatibility of graphene-based supports, but the biological interactions between graphene and neurons still pose open questions. In this work, the influence of graphene films with different characteristics on the growth and maturation of primary cortical neurons is investigated. Graphene films are grown by chemical vapor deposition progressively lowering the temperature range from 1070 to 650 °C to change the lattice structure and corresponding electrical conductivity. Two graphene-based films with different electrical properties are selected and used as substrate for growing primary cortical neurons: i) highly crystalline and conductive (grown at 1070 °C) and ii) highly disordered and 140-times less conductive (grown at 790 °C). Electron and fluorescence microscopy imaging reveal an excellent neuronal viability and the development of a mature, structured, and excitable network onto both substrates, regardless of their microstructure and electrical conductivity. The results underline that high electrical conductivity by itself is not fundamental for graphene-based neuronal interfaces, while other physico– chemical characteristics, including the atomic structure, should be also considered in the design of functional, bio-friendly templates. This finding widens the spectrum of carbon-based materials suitable for neuroscience applications.European Union (EU) 785219-Graphene Flagship-Core2Ministero degli Affari Esteri e Cooperazione Internazionale of Italy (Farnesina-MAECI) MAE0057294Basic Science Research ProgramCreative Materials Discovery ProgramInternational Research & Development Program through the NRF of Korea 2016M3A7B4910940 2018M3D1A1058793 2019K1A3A1A25000267European Union's Horizon 2020 under the Marie Skodowska-Curie Action-COFUND Athenea3i grant 754446European Union's Horizon 2020 research and innovation program under the Marie Skodowska-Curie grant 71364

Multidisciplinary screening of toxicity induced by silica nanoparticles during sea urchin development

The aim of this study was to investigate the potential toxicity of Silica nanoparticles (SiO2 NPs) in seawater by using the sea urchin Paracentrotus lividus as biological model. SiO2 NPs exposure effects were identified on the sperm of the sea urchin through a multidisciplinary approach, combining developmental biology, ecotoxicology, biochemistry, and microscopy analyses. The following responses were measured: (i) percentage of eggs fertilized by exposed sperm; (ii) percentage of anomalies and undeveloped embryos and larvae; (iii) enzyme activity alterations (acetylcholinesterase, AChE) in the early developmental stages, namely gastrula and pluteus. Sperms were exposed to seawater containing SiO2 NPs suspensions ranging from 0.0001 mg/L to 50 mg/L. Fertilization ability was not affected at any concentration, whereas a significant percentage of anomalies in the offspring were observed and quantified by means of EC50 at gastrula stage, including undeveloped and anomalous embryos (EC50 = 0.06 mg/L), and at pluteus stage, including skeletal anomalies and delayed larvae (EC50 = 0.27 mg/L). Moreover, morphological anomalies were observed in larvae at pluteus stage, by immunolocalizing molecules involved in larval development and neurotoxicity effects \u2013 such as acetylated tubulin and choline acetyltransferase (ChAT) \u2013 and measuring AChE activity. Exposure of sea urchins to SiO2 NPs caused neurotoxic damage and a decrease of AChE expression in a non-dose-dependent manner. In conclusion, through the multidisciplinary approach used in this study SiO2 NPs toxicity in sea urchin offspring could be assessed. Therefore, the measured responses are suitable for detecting embryo- and larval- toxicity induced by these NPs