阅读说明：本技术 用于增强脑性能或治疗应激的纳米粒子 (Nanoparticles for enhancing brain performance or treating stress ) 是由 艾格内斯·波迪尔 劳伦特·莱维 马里-艾迪斯·梅尔 于 2018-12-18 设计创作，主要内容包括：本发明涉及医学领域,特别是涉及增强脑性能以及治疗病理性应激。更具体而言,本发明涉及一种纳米粒子或纳米粒子聚集体,其用于增强对象的脑性能或用于预防或治疗对象的病理性应激而无需将所述纳米粒子或纳米粒子聚集体暴露于电场,并优选无需将其暴露于任何其他外部激活源,其中所述纳米粒子或纳米粒子聚集体的材料选自导体材料、半导体材料、介电常数ε<Sub>ijk</Sub>等于或高于(200)的绝缘体材料、和介电常数ε<Sub>ijk</Sub>等于或低于(100)的绝缘体材料。本发明还涉及包含这样的纳米粒子和/或纳米粒子聚集体的组合物和试剂盒,及其在无需将其暴露于电场并优选无需将其暴露于任何其他外部激活源例如光源、磁场或超声源的情况下的用途。(The present invention relates to the field of medicine, and in particular to enhancing brain performance and treating pathological stress. More particularly, the present invention relates to a nanoparticle or aggregate of nanoparticles, wherein the material of the nanoparticle or aggregate of nanoparticles is selected from the group consisting of a conductive material, a semi-conductive material, a dielectric constant material, for enhancing brain performance of a subject or for preventing or treating pathological stress in a subject without exposing the nanoparticle or aggregate of nanoparticles to an electric field, and preferably without exposing the nanoparticle or aggregate of nanoparticles to any other external activation source ijk An insulator material equal to or higher than (200), and a dielectric constant ijk Equal to or lower than (100) insulator material. The invention also relates to compositions and kits comprising such nanoparticles and/or nanoparticle aggregates and their use without exposing them to an electric field and preferably without exposing them to any other external activation source, such as a light source, a magnetic field or an ultrasound source.)

1. A nanoparticle or aggregate of nanoparticles for enhancing brain performance in a subject or for preventing or treating pathological stress in a subject without exposing the nanoparticle or aggregate of nanoparticles to an electric field or any other external activation source, wherein the material of the nanoparticle or aggregate of nanoparticles is selected from the group consisting of a conductive material, a semiconductive material, a dielectric constant_ijkInsulator material of 200 or more, and dielectric constant_ijkAn insulator material equal to or lower than 100, wherein i) when said material is a conductor material, a semiconductor material orDielectric constant_ijkEqual to or higher than 200 of insulator material, the median largest dimension of the cores of the nanoparticles or nanoparticle aggregates of the population is at least 30nm, and wherein ii) the cores of the nanoparticles or nanoparticle aggregates are coated with a biocompatible coating layer providing a neutral or negative surface charge when measured in an aqueous solution with an electrolyte concentration between 0.001 and 0.2M, a nanoparticle or nanoparticle aggregate material concentration between 0.01 and 10g/L and a pH between 6 and 8.

2. Nanoparticle or nanoparticle aggregate for use according to claim 1, wherein the material of the nanoparticle or nanoparticle aggregate is a conductor material selected from metals having a standard reduction potential E ° higher than 0.2 and organic materials having contiguous sp2 hybridized carbon centers in their structure.

3. The nanoparticle or nanoparticle aggregate for use according to claim 2, wherein the material of the nanoparticle or nanoparticle aggregate is selected from metal nanoparticles, wherein the metal element is Ir, Pd, Pt, Au or any mixture thereof, and organic nanoparticles consisting of polyaniline, polypyrrole, polyacetylene, polythiophene, polycarbazole and/or polypyrene.

4. The nanoparticle or nanoparticle aggregate for use according to claim 1, wherein the material of the nanoparticle or nanoparticle aggregate is a semiconductor material with a band gap Eg below 3.0 eV.

5. The nanoparticle or nanoparticle aggregate for use according to claim 4, wherein the material of the nanoparticle or nanoparticle aggregate consists of elements of group IVA of the Mendeleev's periodic Table, or is a mixed composition of elements of groups III and V of the Mendeleev's periodic Table, or is a mixed composition of elements of groups II and VI of the Mendeleev's periodic Table.

6. The nanoparticle or nanoparticle aggregate for use according to claim 5, wherein the material of the nanoparticle or nanoparticle aggregate consists of elements of group IVA of the Mendeleev's periodic Table and is doped with charge carriers selected from Al, B, Ga, In and P.

7. The nanoparticle or nanoparticle aggregate for use according to claim 1, wherein the material is an insulator material with a band gap Eg equal to or higher than 3.0eV and the relative permittivity_ijkBetween 20 ℃ and 30 ℃ and at 10²Hz up to infrared frequencies.

8. The nanoparticle or nanoparticle aggregate for use according to claim 7, wherein said material is an insulator material with a band gap Eg equal to or higher than 3.0eV and said relative permittivity_ijkEqual to or higher than 200, and the material of the nanoparticles or nanoparticle aggregates is a dielectric material selected from BaTiO₃、KTaNbO₃、KTaO₃、SrTiO₃And BaSrTiO₃The mixed metal oxide of (1).

9. The nanoparticle or nanoparticle aggregate for use according to claim 7, wherein said material is an insulator material with a band gap Eg equal to or higher than 3.0eV and said relative permittivity_ijkEqual to or lower than 100 and the material of the nanoparticles or nanoparticle aggregates is selected from ReO₂、ZrO₂And HfO₂The metal oxide of (1).

10. The nanoparticle or nanoparticle aggregate for use according to any one of claims 1 to 9, wherein the nanoparticle or nanoparticle aggregate is for enhancing a physical property of a subject, or enhancing learning, memory, perception, attention and/or decision making of a subject.

11. A composition comprising the nanoparticles and/or nanoparticle aggregates of any one of claims 1 to 9 and a pharmaceutically acceptable carrier, wherein the composition is for enhancing brain performance in a subject or for preventing or treating pathological stress in a subject without exposing the nanoparticles and/or nanoparticle aggregates to an electric field or any other external activation source.

12. The composition for use according to claim 11, wherein the composition comprises at least two different nanoparticles and/or nanoparticle aggregates of any one of claims 1 to 9.

13. A kit comprising at least two different nanoparticles and/or nanoparticle aggregates of any one of claims 1 to 9.

14. The kit of claim 13, for enhancing brain performance in a subject or for preventing or treating pathological stress in a subject without exposing the nanoparticles and/or nanoparticle aggregates to an electric field or any other external activation source.

Background

As understanding of neuroscience advances, the brain can be thought of as an electrical network through which wires, neurons, encode and transmit information. Connectivity between neurons is simple and complex: simply because it consists in the influx/efflux of ions inside the neuron, generating action potentials (or "spikes" of electrical activity); the complexity is due to The fact that brain networks are composed of hundreds of millions of neurons that form nodes, hubs, and modules that exhibit coordinated interactions at various spatial and temporal scales (fortito et al, Nature Reviews Neuroscience, 2015, 16, 159-172: connectivity of brain disorders). Neural communication depends on anatomical components (structures) connecting individual neurons and processes (functions) of transferring information. Both of these aspects affect the overall performance of the nervous system. Oscillations of brain electrical activity patterns, which are typically measurable by electroencephalography (EEG), are detrimental to neuronal interactions. Different oscillation frequency bands are observed: θ, α, β, γ (Ward et al Trends in Cognitive Sciences, 2003, 7(12), 553: (Synchronous neural oscillations and Cognitive processes)). Structurally, the most striking neuroanatomical feature of the brain is the rich connectivity between neurons, which reflects the importance of neural communication. Synchronization of oscillations between one brain region and another ("synchronization") seems to constitute the last stage of information encoding [ first stage (neurons) by introducing space-time coordination: an action potential; second stage (neuronal network): neuronal oscillations ] (Engel et al, Nature Reviews Neuroscience, 2001, 2, 704: 716: Dynamic prediction: oscillations and synchronization in top-down processing (Dynamic predictions: oscillations and synchronization in top-down processing)). Importantly, emerging evidence suggests that a subtle balance of spatial and temporal synchronization and desynchronization patterns is fundamental to the functional performance of the nervous system (Schnitzler et al, Nature reviews neuroscience, 2005, 6, 285; 296: Normal and pathological oscillatory communication in the brain).

The creation of a particular skill, creativity or creative occurrence in some individuals and not others is a very reassuring thing and has not yet been explained. However, studies of certain diseases and their symptoms may be helpful in understanding the function of "normal" and "abnormal" brain. For example, individuals with neurodegenerative diseases such as frontotemporal dementia have been observed to develop mapping and painting skills as their disease progresses (Miller et al, Neurology, 1998, 978-. Several publications indicate that persons working in the creative field (engineering, literature, painting) (and their first degree relatives) suffer from neurological disorders such as bipolar syndrome, the tendency to schizophrenia or Autism Is higher than that of the "non-creators" (Andreaten N.C., American Journal of psychiatric, 1987, 144(10), 1288-1292: Creativity and Mental illness: prevalence in the authors and their first degree relatives (creation and opinion: criteria in and of the first degree relatives), Baron-Cohen et al, Australi, 1997, 101-109: whether or not there Is a link between engineering and Autism (Is thermal link technical and analysis), Sussman et al, Standard Journal of neurological, 2007, 1(1), 21-24: neurological of the "afflicted artist" and creation of Mental illness: "neurological view: neurological of the" afflicted artist "in the name of the" neurological and creation of the "Mental illness". Several models have been elaborated to describe the process of creation and creative generation: hemisphere models, which suggest that the non-dominant hemisphere is dedicated to creative activities, or the most recent Frontotemporal leaf model, which suggests that changes in the temporal leaf may increase the creation of creatives, while changes in the frontal leaf may reduce the creation of creatives (Flaherty et al, J Com Neurol, 2005, 493(1), 147-. Indeed, some scholars may perform esoteric numerical calculations while lacking basic arithmetic (Snyder et al, Proceedings of the Royal Society of London B, 1999, 266, 587. Interestingly, there is evidence that such unusual capacity is associated with left (dominant) hemisphere suppression as well as right (non-dominant) hemisphere facilitation (Treffert D.A., Photocosmetic Transactions of The Royal Society B, 2009, 364, 1351. 1357. scholars syndrome: an unusual condition. Abstract: past, now in The future (The savant syndrome: an extrinsic information. A syndrome: past, present, future)).

The brain is thus a dynamic system in which specific states of brain function arise from complex excitatory and inhibitory interactions between neuronal populations. Thus, an "abnormal" state reflects an imbalance between complex excitatory and inhibitory interactions between neuronal populations (Kapur et al Brain, 1996, 119, 1775; Paradoxical functional promotion in Brain behavior studies, reviewed in (neurological functional assessment in Brain-biochemical research, a clinical review)).

The present invention relates to nanoparticles and/or nanoparticle aggregates (aggregates of nanoparticles) for use in enhancing, increasing or improving brain performance/capacity or for use in preventing or treating pathological stress or at least one symptom thereof.

The nanoparticles and nanoparticle aggregates described herein by the inventors do not require the application/induction of an electrical current or an electrical field/stimulus, and preferably do not require exposure to any other external activation source such as a light source, a magnetic field, or an ultrasound source in order to function (i.e., be effective). The nanoparticles and nanoparticle aggregates described herein do not require exposure to an electric current or electric field/stimulus, and preferably do not require exposure to any other external activation source such as a light source, magnetic field or ultrasound source to be able to function in the context of the uses described herein. The inventors have found that these nanoparticles or nanoparticle aggregates can advantageously and surprisingly be effectively used without exposing them or the subject to which they are administered to an electric current or electric field/stimulation, typically an electric current or electric field/stimulation applied to said subject, e.g. by Transcranial Electrical Stimulation (TES) or Transcranial Magnetic Stimulation (TMS), and preferably without exposing to any other external activation source, such as a light source, a magnetic field or an ultrasound source. This means that, thanks to the present invention, the treated subject will not suffer from the negative side effects of exposure to electric currents or electric fields/stimuli or any other external activation source, such as a light source, a magnetic field or an ultrasound source.

As is well known to those skilled in the art, nanoparticles have an enhanced/high surface area to volume ratio, typically about 35-40% of the atoms of a 10nm nanoparticle are located on the surface, compared to less than 20% of nanoparticles having a size above 30 nm. This high surface area to volume ratio is associated with the strong surface reactivity which is size dependent. Thus, nanoparticles (especially nanoparticles smaller than 20 nm) may exhibit novel properties compared to bulk materials. For example, gold particles are known to be chemically inert and resistant to oxidation on a macroscopic scale, while gold particles below 10nm in size have a chemically active surface. The toxicity mechanisms associated with Chemical destabilization of metal nanoparticles may be (i) the direct release of the metal in solution (dissolution process), (ii) the catalytic properties of the metal nanoparticles, and (iii) the redox evolution of the nanoparticle surface, which can oxidize proteins, generate Reactive Oxygen Species (ROS), and induce oxidative stress (see m.affan et al, Environmental Pollution 157(2009) 1127-1133: Chemical Stability of the metal nanoparticles: parameters for controlling their potential cytotoxicity in vitro (Chemical Stability of metallic nanoparticles: a parameter controlling the potential cytotoxicity of cellular toxicity in vitro)).

Cerium oxide (7nm — CeO) in addition to the gold nanoparticles exhibiting catalytic properties described above₂Particles) or iron oxide (20 nm-Fe)₃O₄Particle) nanoparticles also show redox modification of their surface, leading to cytotoxic effects associated with oxidative stress in vitro (see m.affan et al, Environmental Pollution 157(2009) 1127-1133: chemical stability of metal nanoparticles: parameters for controlling the potential cytotoxicity in vitro (Chemical Stability of metallic nanoparticles: a parameter controlling the cytotoxic activity in vitro)). Likewise, 11 nm-silicA nanostructures are also attacked by biological mediA (see S-A Yang et alHuman, Scientific Reports 20188: 185: the stability of Silica nanoparticles in biological media was reviewed (Silica nanoparticle stability in biological media).

Thus, as the inventors hereinafter explain, when intended for in vivo use in a subject, typically a mammal, in particular a human, nanoparticles having a size below 30nm are carefully selected.

Disclosure of Invention

Advantageously described herein for the first time are nanoparticles or nanoparticle aggregates for enhancing, increasing or improving brain performance/ability of a subject or for preventing or treating a pathological stress or at least one symptom thereof in a subject without exposing said nanoparticles or nanoparticle aggregates to an electric field, and preferably without exposing them to any other external activation source, such as a light source, a magnetic field or an ultrasound source. The material of the nanoparticle or nanoparticle aggregate is typically selected from the group consisting of a conductive material, a semiconductive material, a dielectric constant_ijkInsulator material of 200 or more, and dielectric constant_ijkInsulator material equal to or lower than 100.

In one particular aspect, the inventors describe herein a nanoparticle or aggregate of nanoparticles for enhancing brain performance of a subject or for preventing or treating pathological stress in a subject without exposing the nanoparticle or aggregate of nanoparticles to an electric field or any other external activation source, wherein the material of the nanoparticle or aggregate of nanoparticles is selected from the group consisting of a conductive material, a semiconductive material, a dielectric constant material_ijkInsulator material of 200 or more, and dielectric constant_ijkAn insulator material equal to or lower than 100, wherein i) when said material is a conductor material, a semiconductor material or a dielectric constant_ijkEqual to or higher than 200 of insulator material, the median largest dimension of the cores of the nanoparticles or nanoparticle aggregates of the population is at least 30nm, and wherein ii) the cores of said nanoparticles or nanoparticle aggregates are coated with a biocompatible coating, when at an electrolyte concentration between 0.001 and 0.2M, a nanoparticle or nanoparticle aggregate material concentration between 0.01 and 10g/L and a pH between 6 and 10g/L8, the coating provides a neutral or negative surface charge when measured in an aqueous solution.

Also described herein is the use of a nanoparticle or aggregate of nanoparticles for the preparation of a composition for enhancing, increasing or improving brain performance/capacity or for preventing or treating pathological stress or at least one symptom thereof in a subject in need thereof without exposing the nanoparticle or aggregate of nanoparticles to an electric field, and preferably without exposing it to any other external activation source, such as a light source, a magnetic field or an ultrasound source.

Also described herein are compositions for enhancing brain performance in a subject or for preventing or treating pathological stress or at least one symptom thereof in a subject, wherein the composition comprises or consists of a nanoparticle and/or aggregate of nanoparticles and a pharmaceutically acceptable carrier, wherein the material of the nanoparticle or aggregate of nanoparticles is typically selected from the group consisting of a conductive material, a semiconductive material, a dielectric constant, and a pharmaceutically acceptable carrier_ijkInsulator material of 200 or more, and dielectric constant_ijkAn insulator material equal to or lower than 100, and wherein said enhancing brain performance or preventing or treating pathological stress is performed without exposing the nanoparticles or nanoparticle aggregates administered to said subject by said composition to an electric field and preferably without exposing it to any other external activation source such as a light source, a magnetic field or an ultrasound source.

Also described herein are kits comprising or consisting of at least two different nanoparticles and/or nanoparticle aggregates, each nanoparticle or nanoparticle aggregate consisting of a material generally selected from the group consisting of a conductive material, a semiconductive material, a dielectric constant_ijkInsulator material of 200 or more, and dielectric constant_ijkA different material composition of the insulator material equal to or lower than 100, and said kit in general without exposing said nanoparticles or nanoparticle aggregates to an electric field and preferably without exposing them to any other external activation source such as a light source, a magnetic field or an ultrasound sourceIn or in a method of enhancing brain performance in a subject, or in preventing or treating pathological stress or at least one symptom thereof in a subject.

Detailed Description

The Human nervous system is estimated to consist of approximately 800-. A defining characteristic of a neuron (or nerve cell) is its ability to transmit electrical signals in the form of action potentials.

Neurons/nerve cells constitute the fundamental nodes of the brain. The structure of neurons/nerve cells consists of: "body" or "cell body" which contains a nucleus and is extendible by dendrites; an "axon," which transmits electrical signals; and axon terminals, which consist of synaptic terminals.

Neural cells can communicate with each other in a highly structured manner, forming a network of neurons. Neurons communicate via synaptic connections. Within neurons, the nanocircuit constitutes the basic biochemical mechanism for mediating the occurrence of key neuronal properties such as learning and memory, and neuronal rhythmicity.

Only a few interconnected neurons can form a microcircuit and can perform complex tasks such as mediating reflexes, processing sensory information, initiating motor, and learning and memory mediation. A macro-loop is a more complex network composed of multiple embedded micro-loops. The macrocircuit mediates higher brain functions such as object recognition and cognition. Thus, the multi-stage network occupies the nervous system.

Excitability of neural network

Neurons send messages electrochemically (i.e., chemicals/ions cause electrical signals). Important ions in the nervous system are sodium and potassium, calcium and chloride. When a neuron does not send a signal, it is "quiescent". When a neuron is at rest, the interior of the neuron is negative with respect to the exterior. Although the concentrations of different ions attempt to equilibrate on both sides of the membrane, they cannot because the cell membrane allows only some ions to pass through the channel (ion channel). In addition to these selective ion channels, there are pumps that use energy to move three sodium ions out of the neuron for every two potassium ions that are put in. Finally, when all these forces are balanced and the voltage difference between the interior and exterior of the neuron is measured, the resting membrane potential (also called "resting potential") of the neuron is about-70 mV. This means that the interior of the neuron is 70mV lower than the exterior. At rest, there are relatively many sodium ions outside the neuron and relatively many potassium ions inside the neuron. Action potentials (also identified as "spikes" or "pulses") occur when neurons send information along axons away from the cell body. This means that certain events (stimuli) cause the resting potential to move towards 0 mV. When depolarization reaches approximately-55 mV, the neuron fires an action potential. If the depolarization does not reach the critical threshold level, no action potential is emitted (on/off mechanism). In addition, when a threshold level is reached, an action potential of fixed amplitude is always delivered. Thus, either depolarization does not reach the threshold or a full action potential is generated.

A large variability in the propagation velocity of the action potential was found. In practice, the propagation velocity of action potentials in nerves can vary from 100 m/s to less than one tenth of a m/s. However, the time constant is an indicator of how quickly the membrane responds to the stimulus in time, and the spatial constant (also the length constant) is an indicator of how well the potential spreads along the axon, as a function of distance.

Structure of cerebral cortex

Cortical neurons are of two major classes: "inhibitory neurons" or "interneurons" which produce only short-range local connections; and "excitatory neurons" or "projection neurons" or "pyramidal neurons" that extend axons to distant intra-cortical, sub-cortical and sub-cerebral targets. "inhibitory neurons" or "interneurons" account for a minority of cortical neurons (20%); most are contained in "pyramidal neurons" (Shipp S., Current Biology, 2007, 17(12), R443-449: Structure and function of the cerebral cortex (Structure and function of the cerebral cortex)). Projection neurons are glutamatergic neurons that transmit information between different regions of the neocortex and to other regions of the brain (Bikson et al, J Physiol, 2004, 557(1), 175- & 190: the effect of a uniform extracellular DC electric field on excitability in vitro rat hippocampal slices (Effects of inorganic extracellular DC electric fields on excitability in rat hippocampus slices in vitro)). The projection or pyramidal neurons are named for their protruding apical dendrites, which are usually directed towards the surface layer, giving them a pyramidal morphology. Typically, neurons "belong" to the layer in which their cell bodies (or "bodies") reside-even though they span several layers between apical and basal dendrites, they collect a wider range of signals (Shipp s., Current Biology, 2007, 17(12), R443-449: Structure and function of the cerebral cortex (Structure and function of the nuclear cortex)).

The gray matter of the cerebral cortex is a convoluted piece of stratified tissue in the human body, 2-3 mm thick, but with a surface area of several hundred square centimeters (Shipp s., Current Biology, 2007, 17(12), R443-449: Structure and function of the cerebral cortex (Structure and function of the nuclear cortex)). Six major layers are identified in the cerebral cortex:

layer I, a molecular layer, containing few scattered neurons and consisting mainly of apical dendrite clusters of pyramidal neurons and extensions of horizontally oriented axons and glial cells;

layer II, the outer granular layer, mainly containing small and medium pyramidal neurons and numerous astroid neurons;

layer III, the outer pyramidal cell layer, contains mainly small and medium pyramidal neurons, and non-pyramidal neurons with perpendicularly oriented intra-cortical axons;

layer IV, the inner granular layer, contains different types of star and pyramidal neurons;

layer V, the inner pyramidal cell layer, contains large pyramidal neurons that produce axons that leave the cortex and descend down to sub-cortical structures (e.g., the basal ganglia). In the primary motor cortex of the frontal lobe, layer V contains cells whose axons pass through the inner capsule, the brainstem and the spinal cord forming the corticospinal tract, which is the main pathway of voluntary motor control; and

layer VI, a polymorphic or polytopic layer, containing few large pyramidal neurons and many small spindle pyramidal neurons and pleomorphic neurons; layer VI sends the efferent fibers to the thalamus, establishing a very precise reciprocal interconnection between the cortex and thalamus.

These layers develop differently in various regions of the cerebral cortex, for example, the pyramidal cell layer is more developed in the motor center of the cerebral cortex and the granular layer is more developed in the sensory center.

Connectivity within and between neural networks

There are three types of connectivity networks used to study intra-and whole-brain communications. Structural connectivity is based on the detection of fiber tracks that physically connect brain regions. These are anatomical network maps that indicate the possible ways in which signals may travel in the brain. Functional connectivity identifies activity in brain regions with related activity of similar frequency, phase, and/or amplitude. Effective connectivity uses functional connectivity information and further determines the direct or indirect impact one nervous system may have on another, more specifically the direction of dynamic information flow in the brain (Bowyer et al, Neuropsychiatric Electrical, 2016, 2(1), 1-12: Coherence-a measure of the brain network: past and present (Coherence a measure of the brain network: past and present)).

Synchronous activity within a neuronal network can be detected by Magnetoencephalography (MEG), electroencephalography (EEG), Functional Magnetic Resonance Imaging (FMRI), or Positron Emission Tomography (PET), and then imaged using network connectivity analysis. MEG (magnetoencephalogram) or EEG (electroencephalogram) are preferred because they have high temporal resolution to resolve dynamic information flow. Connectivity analysis of the brain is performed to map out the communication networks required for the brain to function. Specific areas in the brain are dedicated to processing certain types of information. Imaging techniques have revealed that these areas connect and communicate with other specialized areas throughout the network in the brain. "coherence" (Bowyer et al) is a mathematical technique that quantifies the frequency and amplitude of synchronicity (in a synchronized or in-sync state) of neuronal patterns of oscillatory brain activity. Detecting the synchronous activation of neurons can be used to determine the health or integrity of functional connectivity in the human brain. Superimposing a functional connectivity map onto a structural connectivity image and exploiting the direction of information flow obtained from the effective connectivity provides a comprehensive understanding of how the brain functions.

The intact (i.e., "normal" or "healthy") brain expresses complex (i.e., "normal" or "healthy") patterns of synchronized activity, associated with different 'states' of the organism, ranging from slow rhythms (0.5-4Hz), to θ (4-8Hz), α (8-12Hz), β (15-30Hz), and γ (30-70Hz) oscillations. Interestingly, the scatter culture of cortical structures provides a convenient system for examining the regulatory rules for the occurrence, generation and propagation of network firing (spikes) and outbreaks (spike clusters) in densely interconnected neuronal populations. Network activity can be recorded for long periods of time in a non-invasive manner and with limited time resolution using a multi-electrode array. The two-dimensional scatter culture can be used as a viable test system for studying regulatory rules for the formation and maintenance of network activity in the Brain, allowing testing of unresolved hypotheses in the intact Brain (Cohen E. et al Brain Research, 2008, 1235, 21-30: Determinants of spontaneous activity in the cultured hippocampal network (deterinal of specific activity in networks of small hippopathic animals)).

The mental abilities or brain performance, such as intelligence, of humans are particularly complex. Understanding these capabilities in terms of mechanism has the potential to facilitate their enhancement. Studies using electroencephalography and event-related potentials have shown that the speed and reliability of neurotransmission is associated with higher performance, and generally higher intelligence. Early neuroimaging studies using PET found that intelligence during mental activity was negatively associated with brain glucose metabolism, leading to the formulation of a "neural efficiency" hypothesis. According to this hypothesis, a smarter individual spends less neuronal resources at a given level of performance. Intelligence in the sense of reasoning and new problem solving capabilities is always associated with the integrity, structure and function of the lateral prefrontal cortex and possibly with the integrity, structure and function of other areas. The pending questions about the neural basis of intelligence include, among others, the intelligence of psychology tests (i.e. measured by IQ-type tests, usually to assess the accuracy (but not the speed) of the response) and the relationship between: (i) functional connectivity between the components of the working memory network, as demonstrated by electroencephalography-based studies, and (ii) neuroplasticity (i.e., those processes used to refer to the major associative changes involved in the nervous system's response to experience and those processes observed to cease functioning as the human matures). It has been reported that the development of neural junctions is consistent with the development of intelligence (Gray J.R. et al, Nature Review Neuroscience, 2004, 5, 471-.

Communication between neurons is indeed essential for higher brain functions such as perception, memory and movement (Massobrio P et al Neural plastics, 2015, article ID196195, In vitro studies of neuronal networks and synaptic Plasticity In invertebrates and mammals using multi-electrode arrays (In vitro students of Neural networks and synthetic plastics In invertebrates and In mammalian using multiple arrays)). Although the formation and development of linkages are thought to be critical in the learning process, their conservation appears to be essential for memory. Synaptic plasticity has long been implicated in cognitive processes such as learning and memory. Synaptic plasticity at the network level provides a distributed mechanism to convert and store temporal information into a spatially distributed pattern of synapse modifications. Each time something is learned, the network develops new connectivity and incorporates the newly learned facts.

The effective connections between neurons can generally be detected using imaging methods well known to the skilled person, such as electron-based imaging methods that provide structural information about synaptic connectivity, typically Electron Microscopy (EM), such as continuous block-surface electron microscopy (SBFEM), continuous slice scanning electron microscopy (SSSEM), automatic transmission EM (atem), etc.; photon-based imaging Methods, such as "brain rainbow" (Lichtman JW et al, Curr Opin Neurobiol, 2008, 22, 144-; omics: genomics for omics relativity: what information we can tell about the connected groups a direct synapse amplification, genomic imaging Methods and markers), cross-synaptic partner GFP reconstitution ("GRASP"), in particular mammalian GRASP "mGRASP" (Kim J et al, 2012, Nat Methods, 9(1), 96-102: mGRASP enables mapping of mammalian synaptic connectivity with light microscopy using optical microscopy; feng L et al, 2012, Bioinformatics, 28, i25-i 31: improved synapse detection for mGRASP-assisted brain connectivity (Improved synapse detection for mGRASP-assisted brain connectivity)), transsynaptic tracking by rabies virus (Osakada F et al, 2011, Neuron, 71, 617-631: new rabies virus variants (New viruses variants for monitoring and manipulating activity and regulatory activity and gene expression in defined neural circuits); wickersham IR et al, 2007, Nat Methods, 4(1), 47-49: retrograd neuronal tracking with a deletion-mitant rabis virus with a deletion mutant rabies virus; wickersham IR et al, 2007, Neuron, 53(5), 639-: single synaptic constraints (Monosynaptic restriction of transsynaptic tracking from single, generational targeted nerves) that follow transsynaptic from a single gene-targeted neuron, fluorescence selective planar illumination microscopy (fsppim) (Tomer R et al, 2012Natmethods, 9, 755-: quantitative high-speed imaging of the whole developing embryo with simultaneous multi-viewpoint light sheet illumination microscopy (Quantitative high-speed imaging of incident measuring embryo with multiple views-skin microscopy); york AG et al, 2012, Nat Methods, 9(7), 749-: resolution is doubled in living multicellular organisms by illumination microscopy of multifocal structures (Resolution doubling in live, multicellular organization of via multicellular transmitted microscopy), preferably in combination with a transparentizing method such as "gradient" (Chung K et al, 2013, Nature, 497(7449), 332-: structural and molecular interrogation of intact biological systems (Structural and molecular interaction of interactive biological systems)); and optogenetic methods such as channel-rhodopsin and/or two-photon microcalcification imaging methods which allow mapping the spatial distribution of synaptic connections and measuring synaptic strength (Petrenuu L et al, 2007, Nat Neurosci, 10, 663: (Channelrhodopsin-2-assisted circuit mapping of long-range corpus callosum projection); Wang H et al, 2007, Proc Natl Acad Sci USA, 104, 8143: (High-speed mapping of synthetic connective tissue trans2) in channel rhodopsin-2transgenic mice) and detecting activity of synapses with different inputs (JP-High-speed mapping of synthetic connective tissue synuclein-2) in channel rhodopsin-2transgenic mice) (32: 32, 32: neuron J, 32, 2. fig., 12808-12819: subcellular synaptic connectivity of layer2pyramidal neurons in the medial prefrontal cortex (Subcellular synaptic connectivity of layer2pyramidal neurons in the medial prefrontal cortex); MacAskill AF et al, 2012, NatNeurosci, 15(12), 1624-: subcellular connectivity is the basis for nucleus accumbens pathway-specific signaling (subellular connectivity assays in the nucleus); or any combination of these different methods (Yook C. et al, Cellular and Molecular Life sciences, 2013, 70, 4747) 4757: Mapping mammalian synaptic connectivity).

Advantageously described herein for the first time are nanoparticles or nanoparticle aggregates for enhancing, increasing or improving brain performance/capacity or for preventing or treating pathological stress or at least one symptom thereof without exposing said nanoparticles or nanoparticle aggregates to an electric field, and preferably without exposing them to any other external activation source, such as a light source, a magnetic field or an ultrasound source. Such exposure to a (therapeutic or diagnostic) electric field or any other (therapeutic or diagnostic) external activation source, such as a light source, a magnetic field or an ultrasound source, is generally understood herein as a therapeutic or diagnostic exposure, typically performed by medical personnel, e.g. by a doctor or nurse.

The material of the nanoparticle or nanoparticle aggregate is typically selected from the group consisting of a conductive material, a semiconductive material, a dielectric constant_ijkInsulator material of 200 or more, and dielectric constant_ijkInsulator material equal to or lower than 100.

In a typical aspect, the nanoparticles or nanoparticle aggregates described herein are used to enhance a subject's physical performance, or to enhance a subject's cognitive performance, i.e. learning, memory, perception, attention and/or decision making, without exposing said nanoparticles or nanoparticle aggregates to an electric field, and preferably without exposing them to any other external activation source, such as a light source, a magnetic field or an ultrasound source.

Nanoparticles

Described herein is a nanoparticle or aggregate of nanoparticles for use according to the invention for enhancing brain performance of a subject or for treating a pathological stress or at least one symptom thereof in a subject without exposing the nanoparticle or aggregate of nanoparticles to an electric field, and preferably without exposing the nanoparticle or aggregate of nanoparticles to any other external activation source, such as a light source, a magnetic field or an ultrasound source, wherein the nanoparticle or aggregate of nanoparticles isThe material of the seed or nanoparticle aggregate is generally selected from the group consisting of a conductive material, a semiconductive material, a dielectric constant_ijkInsulator material of 200 or more, and dielectric constant_ijkInsulator material equal to or lower than 100.

Size or dimensions of nanoparticles or nanoparticle aggregates

In the spirit of the present invention, the term "nanoparticle" or "nanoparticle aggregate" refers to a product, in particular a synthetic product, having a size in the nanometer range, typically between 1nm and 1000nm, or between 1nm and 500nm, for example between at least 10nm and about 500nm or about 1000nm, between at least 30nm and about 500nm or about 1000nm, between at least 40nm and about 500nm or about 1000nm, between at least 45nm and about 500nm or about 1000nm, preferably below 500 nm.

The term "aggregate of nanoparticles" or "nanoparticle aggregate" refers to an assembly of nanoparticles that are strongly, usually covalently, bound to each other.

Electron microscopy, such as Scanning Electron Microscopy (SEM), Transmission Electron Microscopy (TEM) or cryotem, may be used to measure the size of the nanoparticles or nanoparticle aggregates, more particularly the size of the core of the nanoparticles or nanoparticle aggregates, i.e. the size of the nanoparticles or nanoparticle aggregates without the biocompatible coating. In fact, said biocompatible coating is generally made of compounds (polymeric or organic compounds) mainly composed of light elements, which have a relatively weak elastic interaction with high-energy electrons, resulting in poor image contrast. TEM measures the projected image of particles deposited on an electron transparent substrate. More than 50, preferably more than about 100, 150 or 200 nanoparticles or nanoparticle aggregates recorded per sample should generally be subjected to size assessment measurements. Thus, recording more than about 50, or preferably more than about 100, 150 or 200 nanoparticles or nanoparticle aggregates allows determining the median maximum size of the cores of the nanoparticles or nanoparticle aggregates of the population, and the size of the cores of the nanoparticles or nanoparticle aggregates representing 30% -70% of the percentile of the population of nanoparticles or nanoparticle aggregates. Typical assay protocols can be found in the "NIST-NCL combined assay protocol, PCC-7; measuring dimensions using Transmission Electron Microscopy (TEM); version 1.1 at 12.2009 (NIST-NCL Joint Assay Protocol, PCC-7; Measuring the size of using Transmission Electron Microscopy (TEM); version 1.1 December 2009) ".

Likewise, Dynamic Light Scattering (DLS) can also be used to measure the hydrodynamic diameter of a nanoparticle or nanoparticle aggregate in solution (i.e., the diameter of the nanoparticle or nanoparticle aggregate including both its core and its biocompatible coating). The hydrodynamic diameter is the diameter of an equivalent hard sphere that diffuses at the same rate as the analyte. Typical assay protocols can be found in the "NIST-NCL combined assay protocol, PCC-1; measuring the size of nanoparticles in an aqueous medium using batch mode dynamic light scattering; version 1.1 at 2.2010 (NIST-NCL Joint Assay Protocol, PCC-1; Measuring the size of nanoparticles in aqueous media using batch-mode dynamic lighting, version 1.1 February 2010). The particle size results obtained from DLS measurements may not be consistent with those obtained from other techniques (e.g., electron microscopy). This is due in part to the difference in the physical properties (e.g., hydrodynamic diffusion and projected area) that are actually measured. Furthermore, DLS is sensitive to the presence of small numbers of large or small particle clusters, whereas electron microscopy generally reflects the size of the primary particles (i.e., the size of the core of the nanoparticle or nanoparticle aggregate) (see NIST-NCL Combined Assay Protocol, PCC-1; measurement of the size of nanoparticles in aqueous media using batch mode dynamic light scattering; version 1.1, 2.2010 (NIST-NCL Joint Assay Protocol, PCC-1; measurement of the size of nanoparticles in aqueous media using a batch mode dynamic light scattering; version 1.1 February 2010)).

Both DLS and electron microscopy methods can also be used one after the other to compare dimensional measurements and confirm the dimensions. A preferred method for measuring the Size of nanoparticles and nanoparticle aggregates is DLS (see International Standard ISO22412Particle Size Analysis-Dynamic Light Scattering, International organization for standardization (ISO)2008(International Standard ISO22412Particle Size Analysis-Dynamic Light Scattering, International organization for standardization (ISO) 2008)). The average hydrodynamic diameter of the nanoparticles or nanoparticle aggregates measured by DLS in solution is given as a size distribution according to intensity (light scattering intensity is proportional to particle size) and is measured at room temperature (about 25 ℃).

Typically, the largest dimension or size is the diameter of a round or spherical shaped nanoparticle, or the longest length of an oval or elliptical shaped nanoparticle.

The largest dimension of a nanoparticle or aggregate as defined herein is typically between about 2nm and about 250nm or about 500nm, preferably between about 4nm or 10nm and about 100nm or about 200nm, more preferably between about (preferably at least) 10nm and about 150nm, between about (preferably at least) 30nm and about 150nm, between about (preferably at least) 40nm and about 500nm, between about (preferably at least) 45nm and about 500nm, preferably below 500 nm.

DLS techniques are typically used when measuring the average hydrodynamic diameter of a nanoparticle or nanoparticle aggregate in solution. Using DLS, the average hydrodynamic diameter of the nanoparticle or nanoparticle aggregate in solution is typically between about 10nm and about 500nm, preferably between about 10nm or about 30nm and about 100nm or about 500nm, more preferably between about 10nm or about 30nm and about 100nm, about 150nm, about 200nm, about 250nm, about 300nm, about 350nm, about 400nm, about 450nm or about 500 nm.

When measuring the core of a nanoparticle or nanoparticle aggregate, electron microscopy techniques are typically used. Using electron microscopy, the median largest dimension (also referred to herein as the "median largest dimension") of the cores of the nanoparticles or nanoparticle aggregates of the population is typically between about 5nm and about 250nm or about 500nm, preferably about 5nm, about 6nm, about 7nm, about 8nm, about 9nm, about 10nm, about 11nm, about 12nm, about 13nm, about 14nm, about 15nm, about 16nm, about 17nm, about 18nm, about 19nm, about 20nm, about 21nm, about 22nm, about 23nm, about 24nm, about 25nm, about 26nm, about 27nm, about 28nm, about 29nm, about 30nm, about 31nm, about 32nm, about 33nm, about 34nm, about 35nm, about 36nm, about 37nm, about 38nm, about 39nm, about 40nm, about 41nm, about 42nm, about 43nm, about 44nm or about 45nm, about 75nm, about 76nm, about 78nm, about 79nm, about 27nm, about 28nm, about 29nm, about, About 81nm, about 82nm, about 83nm, about 84nm, about 85nm, about 86nm, about 87nm, about 88nm, about 89nm, about 90nm, about 91nm, about 92nm, about 93nm, about 94nm, about 95nm, about 96nm, about 97nm, about 98nm, about 99nm, about 100nm, about 101nm, about 102nm, about 103nm, about 104nm, about 105nm, about 106nm, about 107nm, about 108nm, about 109nm, about 110nm, about 111nm, about 112nm, about 113nm, about 114nm, about 115nm, about 116nm, about 117nm, about 118nm, about 119nm, about 120nm, about 121nm, about 122nm, about 123nm, about 124nm, about 125nm, about 130nm, about 140nm, about 150nm, about 200nm, about 250nm, about 300nm, about 350nm, about 400nm, about 450nm, or about 500 nm.

Typically, when the size of the core of a nanoparticle or aggregate of nanoparticles is measured with an electron microscopy instrument, the size of the core of the nanoparticle or aggregate of nanoparticles, representing a 30% -70% percentile of the population of nanoparticles and aggregates of nanoparticles, is between about 5nm, about 6nm, about 7nm, about 8nm, about 9nm, about 10nm, about 11nm, about 12nm, about 13nm, about 14nm, about 15nm, about 16nm, about 17nm, about 18nm, about 19nm, about 20nm, about 21nm, about 22nm, about 23nm, about 24nm, about 25nm, about 26nm, about 27nm, about 28nm, about 29nm, about 30nm, about 31nm, about 32nm, about 33nm, about 34nm, about 35nm, about 36nm, about 37nm, about 38nm, about 39nm, about 40nm, about 41nm, about 42nm, about 43nm, about 44nm or about 45nm, about 75nm, about 76nm, about 77nm, About 78nm, about 79nm, about 80nm, about 81nm, about 82nm, about 83nm, about 84nm, about 85nm, about 86nm, about 87nm, about 88nm, about 89nm, about 90nm, about 91nm, about 92nm, about 93nm, about 94nm, about 95nm, about 96nm, about 97nm, about 98nm, about 99nm, about 100nm, about 101nm, about 102nm, about 103nm, about 104nm, about 105nm, about 106nm, about 107nm, about 108nm, about 109nm, about 110nm, about 111nm, about 112nm, about 113nm, about 114nm, about 115nm, about 116nm, about 117nm, about 118nm, about 119nm, about 120nm, about 121nm, about 122nm, about 123nm, about 124nm, about 125nm, about 130nm, about 140nm, about 150nm, about 200nm, about 250nm, about 300nm, about 350nm, about 520nm, about 500nm, or between.

Composition of nanoparticles

Nanoparticles made from conductive materials

The nanoparticles prepared from the conductive material are organic nanoparticles or inorganic nanoparticles.

Inorganic nanoparticles prepared from conductive materials are typically prepared with metallic elements having a standard reduction potential E ° value of equal to or higher than about 0.01 (see table 2 "reduction reactions with E ° values more positive than standard hydrogen electrodes", 8-25, Handbook of chemistry and physics (David r.lide; 88 th edition), more preferably equal to or higher than about 0.1, 0.2, 0.3, 0.4 or 0.5, when measured against standard hydrogen electrodes, typically at 25 ℃ and 1atm pressure. Typical metal elements used to prepare the nanoparticles may be selected from Tl, Po, Ag, Pd, Ir, Pt, Au, and mixtures thereof. Preferably, the metal element that can be used as conductor material for preparing the nanoparticles is selected from Ir, Pd, Pt, Au, and mixtures thereof, more preferably from Au, Pt, Pd, and mixtures thereof. Particularly preferred materials are Au and Pt.

Generally, gold nanoparticles exhibit catalytic activity when their size is reduced to a few nanometers (see M.Auffan et al, Nature Nanotechnology 2009, 4(10), 634-641: the definition of inorganic nanoparticles from an environmental, health and safety perspective (labor a definition of inorganic nanoparticles from environmental and safety). In order to reduce the surface area to volume ratio and thereby minimize the contribution of the inorganic nanoparticle surface to the catalytic activity, it is preferred that the median largest dimension of the cores of the nanoparticles or nanoparticle aggregates of the population is at least 30nm, typically at least 40nm or at least 45 nm.

Organic nanoparticles prepared from conductive materials are typically prepared with organic materials having contiguous sp2 hybridized carbon centers (i.e., carbon double bonds or aromatic rings containing heteroatoms, typically N or S, within or outside the aromatic ring) in their structure. Preferred organic materials are selected from polyaniline, polypyrrole, polyacetylene, polythiophene, polycarbazole, polypyrene, poly (3, 4-ethylenedioxythiophene) and/or poly (3, 4-ethylenedioxythiophene) polystyrene sulfonate.

In a particular aspect, when the material is a conductor material as described above, in particular a metallic material, typically a metal with a standard reduction potential E ° above 0.2, or an organic material, typically an organic material having contiguous sp2 hybridized carbon centers in its structure, preferably a metallic material as described above, in particular any of Au, Pt, Pd and any mixtures thereof, the median largest dimension of the cores of the nanoparticles or nanoparticle aggregates of the population is at least 30nm or at least 40nm and preferably below 500nm, for example 45nm, as described above.

Nanoparticles prepared from semiconductor materials

Nanoparticles prepared from semiconductor materials are typically inorganic nanoparticles.

Inorganic nanoparticles are typically prepared with semiconductor materials that exhibit a relatively small energy bandgap (Eg) between their valence and conduction bands. Typically, the semiconductor material has a bandgap Eg below 3.0eV, when measured typically at room temperature (about 25 ℃) (see, e.g., tables 12-77, table 3; Handbook of chemistry and physics; David r. lite; 88 th edition)). In one particular aspect, the material is intrinsic semiconductor material or extrinsic semiconductor material as described further below.

Intrinsic semiconductor materials are generally composed of elements of group IV A of the Mendeleev (Mendeleev) periodic table, such as silicon (Si) or germanium (Ge), mixed compositions of elements of groups III and V of the Mendeleev periodic table, such as AlSb, AlN, GaP, GaN, InP, InN, etc., or mixed compositions of elements of groups II and VI of the Mendeleev periodic table, such as ZnSe, ZnTe, CdTe, etc.

Extrinsic semiconductor material typically comprises or consists of an intrinsic semiconductor prepared with high chemical purity, wherein the intrinsic semiconductor material comprises a dopant. In a particular aspect, when the extrinsic semiconductor material of the nanoparticle or nanoparticle aggregate consists of an element of group IVA of the mendeleev's periodic table, it is doped with a charge carrier selected from Al, B, Ga, In and P. Such extrinsic semiconductor materials may be n-type, in which negative charge carriers predominate, or p-type, in which positive charge carriers predominate. Typical extrinsic p-type semiconductor materials consist of silicon (Si) or germanium (Ge) doped with a charged carrier selected from aluminum (Al), boron (B), gallium (Ga) and indium (In); typical extrinsic P-type semiconductor materials consist of silicon (Si) or germanium (Ge), usually doped with phosphorus (P).

Generally, the band gap energy of semiconductor nanoparticles shows an increase as the size of the nanoparticles decreases below 10nm (see M.Auffan et al, Nature Nanotechnology 2009, 4(10), 634:. sup. 641. definition of inorganic nanoparticles from environmental, health and safety perspectives). To ensure a low surface area/volume ratio of the nanoparticles or nanoparticle aggregates and to maintain their bulk band gap (bulk band gap) below 3.0eV, it is preferred that the median maximum dimension of the cores of the nanoparticles or nanoparticle aggregates of the population is at least 30nm, preferably at least 40 nm.

Thus, In a particular aspect, when the material is a semiconductor material as described above, In particular a semiconductor material having a band gap Eg below 3.0eV, typically a material consisting of an element of group IVA of the mendeleev periodic table, In particular an element of group IVA of the mendeleev periodic table doped with a charge carrier selected from Al, B, Ga, In and P, or a material consisting of a mixed composition of elements of groups III and V of the mendeleev periodic table, or a material consisting of a mixed composition of elements of groups II and VI of the mendeleev periodic table, the median maximum dimension of the core of the nanoparticles or nanoparticle aggregates of the population is at least 30nm or at least 40nm and preferably below 500 nm.

Nanoparticles prepared from insulator materials having a high relative dielectric constant (relative permittivity), i.e. equal to or higher than 200

Has a high relative dielectric constant_ijk(also known as relative permittivity) of or nanoparticles made of insulator materials are generally prepared with a band gap Eg equal to or higher than 3.0eV and a relative dielectric constant_ijkMaterial preparation equal to or higher than 200, the band gap Eg being generally measured at room temperature (about 25 ℃)And the relative dielectric constant_ijkUsually between 20 ℃ and 30 ℃ and at 10²Measured between Hz and infrared frequencies (see, e.g., tables 12-45, "dielectric constant of inorganic solids (dielectric constant)", "Handbook of chemistry and physics"; David R.Lide; 88 th edition; Compilation of electrostatic dielectric constants of inorganic solids (dielectric constant of inorganic solids), K.F.Young and H.P.R.Fredikse.J.Phys.chem.Ref.Data, Vol.2, No. 2, 1973).

Such nanoparticles are generally prepared with a dielectric material, preferably selected from BaTiO₃、PbTiO₃、KTaNbO₃、KTaO₃、SrTiO₃、BaSrTiO₃And the like.

Typically, a perovskite-based structure of PbTiO₃Nanoparticles exhibit a change in their paraelectric-ferroelectric transition temperature at nanoparticle sizes of less than 20nm to 30nm (see M. Auffan et al, Nature Nanotechnology 2009, 4(10), 634: definition of inorganic nanoparticles from an environmental, health and safety perspective (health and safety)). In order to ensure a low surface area/volume ratio of the nanoparticles or nanoparticle aggregates and maintain their dielectric properties, it is preferred that the median maximum dimension of the cores of the nanoparticles or nanoparticle aggregates of the population is at least 30nm, typically at least 40 nm.

Thus, in one particular aspect, when the material is as described above having a relatively high dielectric constant equal to or higher than 200_ijkIn particular an insulator material having a band gap Eg equal to or higher than 3.0eV, preferably selected from BaTiO₃、KTaNbO₃、KTaO₃、SrTiO₃And BaSrTiO₃In the mixed metal oxide of (a), the median maximum dimension of the cores of the nanoparticles or nanoparticle aggregates of the population is at least 30nm or at least 40nm and preferably below 500 nm.

Nanoparticles prepared from insulator materials having a low relative dielectric constant (relative permittivity), i.e. equal to or lower than 100

Nanoparticles prepared from or consisting of insulator materials having a low relative permittivity are generally prepared with a band gap Eg equal to or higher than 3.0eV and a relative permittivity_ijkEqual to or lower than 100, preferably lower than 50 or lower than 20, said band gap Eg being generally measured at room temperature (about 25 ℃), and said relative dielectric constant_ijkUsually between 20 ℃ and 30 ℃ and 10²Measured between Hz and infrared frequencies (see, e.g., tables 12-45, "dielectric constant of inorganic solids (dielectric constant)", "Handbook of chemistry and physics"; David R.Lide; 88 th edition; Compilation of electrostatic dielectric constants of inorganic solids (dielectric constant of inorganic solids), K.F.Young and H.P.R.Fredikse.J.Phys.chem.Ref.Data, Vol.2, No. 2, 1973).

Such nanoparticles are typically prepared with a dielectric material selected from the group consisting of metal oxides, mixed metal oxides, the metal elements of which are from period 3, 5 or 6 of the mendeleev's periodic table or are lanthanides, and carbon materials. The dielectric material is preferably selected from Al₂O₃、LaAlO₃、La₂O₃、SiO₂、SnO₂、Ta₂O₅、ReO₂、ZrO₂、HfO₂、Y₂O₃And carbon diamond. More preferably, the dielectric material is selected from ReO₂、ZrO₂、HfO₂And any mixtures thereof. Particularly preferred is a material selected from ZrO₂And HfO₂The dielectric material of (1). In a particular and preferred aspect, the dielectric material or metal oxide is not CeO₂(cerium oxide), Fe₃O₄(iron oxide), SiO₂(silica) or any mixture thereof.

Zirconium (Zr) and hafnium (Hf) are both 4⁺Elements in the oxidation state, and Zr⁴⁺And Hf⁴⁺The size and chemical properties of the elements are almost the same; that is why the aqueous chemistry of these two ions is establishedThe reasons for taking them into account (see chapter 8, section 8.2, Zr4+ and Hf4+ (Zr4+ and Hf4+), page 147, "hydrolysis of cations", Baes C.F.&Mesmer r.e.; john Wiley and Sons, Inc.1986 reprinting).

In a particular aspect, as described above, when the material is selected from ReO₂、ZrO₂、HfO₂Preferably selected from ZrO₂And HfO₂And any mixtures thereof, the median maximum dimension of the cores of the nanoparticles or nanoparticle aggregates of the population is at least 10nm and preferably below 500 nm.

Shape of nanoparticles or nanoparticle aggregates

Since the shape of the particles or aggregates may affect their "biocompatibility", particles or aggregates of very uniform shape are preferred. For pharmacokinetic reasons, nanoparticles or aggregates that are substantially spherical, circular or ovoid in shape are therefore preferred. Such a shape also facilitates interaction of the nanoparticle or aggregate with or uptake by cells. A spherical or round shape is particularly preferred.

The shape of the nanoparticle or nanoparticle aggregate is typically evaluated using electron microscopy, such as Transmission Electron Microscopy (TEM).

Biocompatible coating of nanoparticles or nanoparticle aggregates

In a preferred embodiment, the core of the nanoparticle or nanoparticle aggregate used to prepare the subject composition in the context of the present invention may be coated with a biocompatible material selected from agents exhibiting stealth properties. The agent exhibiting stealth properties may be an agent exhibiting a steric group. Such groups may be selected from, for example, polyacrylates; polyacrylamide (poly (N-isopropylacrylamide)); polyamides (polycarboamides); a biopolymer; polysaccharides such as dextran or xylan; and collagen. In another preferred embodiment, the core of the nanoparticle or nanoparticle aggregate may be coated with a biocompatible material selected from agents that allow interaction with biological targets. Such agents may generally carry a positive or negative charge on the surface of the nanoparticle or nanoparticle aggregate. The agent that forms a positive charge on the surface of the nanoparticle or nanoparticle aggregate can be, for example, aminopropyltriethoxysilane or polylysine. The agent forming a negative charge on the surface of the nanoparticle or nanoparticle aggregate may be, for example, phosphoric acid (salt) (e.g., polyphosphoric acid (salt), metaphosphoric acid (salt), pyrophosphoric acid (salt), etc.), carboxylic acid (salt) (e.g., citric acid (salt) or dicarboxylic acid, particularly succinic acid), or sulfuric acid (salt).

In a preferred embodiment, the core of the nanoparticles or nanoparticle aggregates used in the context of the present invention exhibits a hydrophilic neutral surface charge or is coated with a biocompatible material (i.e. a coating agent) selected from hydrophilic agents that impart a neutral surface charge to the nanoparticles. Indeed, when the nanoparticles of the invention are administered to a subject, the core of the nanoparticles presenting a hydrophilic neutral surface charge or of the nanoparticles coated with a biocompatible agent selected from hydrophilic agents that impart a neutral surface charge to the nanoparticles is particularly advantageous for optimizing the use of the nanoparticles described herein.

The hydrophilic agent that imparts a neutral surface charge to the core of the nanoparticle or nanoparticle aggregate may be an agent that exhibits a functional group selected from the group consisting of alcohol (R-OH), aldehyde (R-COH), ketone (R-CO-R), ester (R-COOR), acid (R-COOH), thiol (R-SH), sugar (e.g., glucose, fructose, ribose), acid anhydride (RCOOOC-R), and pyrrole. The hydrophilic agent that imparts a neutral surface charge to the core of the nanoparticle or aggregate of nanoparticles may be a monomer, dimer, oligomer, polymer, or copolymer. When the reagent is an oligomer, it may be an oligosaccharide, for example a cyclodextrin. When the agent is a polymer, it may be a polyester (e.g. poly (lactic acid) or polyhydroxyalkanoic acid), a polyether, polyethylene oxide, polyethylene glycol, polyvinyl alcohol, polycaprolactone, polyvinylpyrrolidone, a polysaccharide such as cellulose, polypyrrole, or the like.

In addition, the hydrophilic agent that imparts a neutral surface charge to the core of the nanoparticle or nanoparticle aggregate may be an agent that exhibits a specific group (R-) capable of interacting with the surface of the nanoparticle or nanoparticle aggregate. R is typically selected from thiol, silane, carboxyl and phosphate groups.

When the core of the nanoparticle or nanoparticle aggregate is a conductor or semiconductor and metal nanoparticle, R is preferably a thiol, thioether, thioester, dithiolane or carboxyl group. Preferably, the hydrophilic neutral capping agent is selected from the group consisting of thioglucose, 2-mercaptoethanol, 1-thioglycerol, thiodiglycol and hydroxybutyric acid.

When the core of the nanoparticle or nanoparticle aggregate is an insulator, and an oxide or mixed oxide nanoparticle, R is preferably a silane or phosphate group. Preferably, the hydrophilic neutral coating agent is hydroxymethyl triethoxysilane, fructose-6-phosphate or a glucose-6-phosphate compound.

The hydrophilic agent that imparts a neutral surface charge to the core of the nanoparticle or aggregate of nanoparticles may be a zwitterionic compound, such as an amino acid, peptide, polypeptide, vitamin, or phospholipid.

As is well known to those skilled in the art, the surface charge of a nanoparticle or nanoparticle aggregate is typically measured by zeta potential, typically determined in water (solution) at a concentration of the nanoparticle or nanoparticle aggregate material of between 0.01 and 10g/L, a pH of between 6 and 8, and a concentration of the electrolyte (in water) of typically between 0.001 and 0.2M, for example 0.01M or 0.15M. The surface charge of the nanoparticle or aggregate of nanoparticles is typically between-10 mV and +10mV (corresponding to a neutral surface charge), -20mV and +20mV, or-35 mV and +35mV, under the conditions defined above. When neutral, the surface charge of the nanoparticle or aggregate of nanoparticles is typically between-10 mV, -9mV, -8mV, -7mV, -6mV, -5mV, -4mV, -3mV, -2mV, or-1 mV and 1mV, 2mV, 3mV, 4mV, 5mV, 6mV, 7mV, 8mV, 9mV, or 10 mV. When negative, the surface charge of the nanoparticle or aggregate of nanoparticles is typically less than-11 mV, -12mV, -13mV, -14mV-15mV, -16mV, -17mV, -18mV, -19mV, -20mV, -21mV, -22mV, -23mV, -24mV, -25mV, -26mV, -27mV, -28mV, -29mV, -30mV, -31mV, -32mV, -33mV, -34mV, or-35 mV.

A biocompatible full coating of the nanoparticles or aggregates may be advantageous in the context of the present invention in order to avoid any charge on the surface of the nanoparticles when the nanoparticles exhibit a hydrophilic neutral surface charge. By "fully coated" is meant the presence of a very high density/compactness of biocompatible molecules capable of producing at least a complete monolayer on the surface of the particle.

The biocompatible coating allows in particular the stability of the nanoparticles in fluids, such as physiological fluids (blood, plasma, serum, etc.) or any isotonic or physiological medium required for drug administration.

Stability can be confirmed by dry extract quantification using a drying oven and measured on the nanoparticle suspension before and after filtration, which is typically performed on a 0.45 μm filter.

Advantageously, the coating maintains the in vivo integrity of the particle, ensures or improves its biocompatibility, and facilitates its optional functionalization (e.g., with spacer molecules, biocompatible polymers, targeting agents, proteins, etc.).

The biocompatible nanoparticles or nanoparticle aggregates of the invention should neither dissolve and release toxic substances after in vivo administration (i.e. at physiological pH) nor exhibit redox behavior, and are generally safe for use in said nanoparticles or nanoparticle aggregates which are considered biocompatible, i.e. in a subject, in particular a mammal, preferably a human.

Another particular object described herein relates to compositions, in particular pharmaceutical compositions, comprising nanoparticles and/or nanoparticle aggregates, such as defined above, preferably together with a pharmaceutically acceptable carrier or medium.

In particular, described herein is a composition for enhancing brain performance of a subject or for preventing or treating pathological stress or at least one symptom thereof in a subject as described herein without exposing the nanoparticles or nanoparticle aggregates to an electric field, and preferably without exposing them to any other external activation source such as a light source, a magnetic field or an ultrasound source, wherein the combinationComprises or consists of nanoparticles and/or nanoparticle aggregates and a pharmaceutically acceptable carrier, and wherein the material of the nanoparticles or nanoparticle aggregates is typically selected from the group consisting of conductor materials, semiconductor materials, dielectric constant materials as described and explained above_ijkInsulator material of 200 or more, and dielectric constant_ijkInsulator material equal to or lower than 100.

In a preferred aspect, the composition comprises or consists of at least two different nanoparticles and/or nanoparticle aggregates, each nanoparticle or nanoparticle aggregate consisting of a different material, typically selected from the group consisting of conductor materials, semiconductor materials, dielectric constant materials_ijkInsulator material of 200 or more, and dielectric constant_ijkInsulator material equal to or lower than 100.

In a typical aspect of the invention, the nanoparticles or nanoparticle aggregates described herein are not used as carriers for (active) therapeutic compounds or drugs.

The composition may be in the form of a solid, liquid (suspended particles), aerosol, gel, paste, or the like. Preferred compositions are in liquid or gel form. Particularly preferred compositions are in liquid form.

The pharmaceutically acceptable carrier or vehicle employed can be any of the classical carriers to the skilled artisan, such as saline, isotonic, sterile, buffered solutions, solutions in non-aqueous media, and the like.

The composition may also include stabilizers, sweeteners, surfactants, polymers, and the like.

It can be formulated, for example, into ampoules, aerosols, bottles, tablets, capsules, by using pharmaceutical formulation techniques known to the skilled worker.

The nanoparticles or nanoparticle aggregates of the invention may be administered to a subject using different possible routes, e.g. intracranial, Intravenous (IV), airway (inhalation), intrathecal, intraocular or buccal (oral), Intracerebroventricular (ICV), preferably using intracranial or intrathecal. Repeated injections or administrations of the nanoparticles or nanoparticle aggregates may be performed, as appropriate. Preferably, the nanoparticle or nanoparticle aggregate is administered once.

The nanoparticles and/or nanoparticle aggregates, once administered, typically interact with the neuronal subject. In a preferred aspect, the interaction is a long-term interaction, i.e., an interaction of hours, days, weeks, or months. In a particular aspect, the nanoparticles or nanoparticle aggregates are left in the subject.

The nanoparticles, nanoparticle aggregates and compositions comprising such nanoparticles or nanoparticle aggregates described herein are for use in a subject, typically for an animal, preferably a mammal, more preferably for a human, regardless of its age or sex.

A typical amount of nanoparticles or nanoparticle aggregates to be administered in the cerebral cortex, hippocampus or amygdala of a subject is in the range of 10⁵And 10¹⁷Between 10⁵And 10¹⁶Between or at 10⁵And 10¹⁵Preferably between 10⁷And 10¹⁴More preferably between 10⁹And 10¹²In the meantime. Also, typical amounts of nanoparticles or nanoparticle aggregates administered in the cerebral cortex, hippocampus or amygdala of a subject are in the range of 10²And 10¹²Nanoparticles or nanoparticle aggregates/cm³In the meantime.

Typical amounts of nanoparticles or nanoparticle aggregates administered in the deep brain of a subject are in the range of 10⁴And 10¹⁷Between 10⁴And 10¹⁶Between 10⁴And 10¹⁵Between or at 10⁴And 10¹⁴Preferably between 10⁶And 10¹²More preferably between 10⁸And 10¹¹In the meantime. And, a typical amount of nanoparticles or nanoparticle aggregates administered in the deep brain of a subject is at 10¹And 10¹¹Nanoparticles or nanoparticle aggregates/cm³In the meantime.

Also described herein is a method of enhancing brain performance in a subject, and a method of treating a subject for pathological stress or at least one symptom thereof, wherein each method comprises the step of administering to the subject any one of the nanoparticles or aggregates of nanoparticles described herein. Such a method typically does not comprise any step of exposing the object, more precisely the nanoparticles or nanoparticle aggregates that have been applied to the object, to an electric field, and preferably also does not comprise any step of exposing the object, more precisely the nanoparticles or nanoparticle aggregates that have been applied to the object, to any other external activation source, such as a light source, a magnetic field or an ultrasound source.

Another object described herein relates to a kit comprising at least two different nanoparticles and/or nanoparticle aggregates as described herein, each nanoparticle or nanoparticle aggregate consisting of a material generally selected from the group consisting of a conductor material, a semiconductor material, a dielectric constant as described herein_ijkInsulator material of 200 or more, and dielectric constant_ijkA different material composition of the insulator material equal to or lower than 100.

In a particular embodiment, the kit comprises in different containers different nanoparticles and/or aggregates of nanoparticles as described herein (which are defined as being contacted, typically mixed, in situ, i.e. at a target site, or in vitro or ex vivo, and then depositing the mixture at the target site).

Also described herein is the use of such a kit in a method of enhancing brain performance/ability, typically enhancing synaptic plasticity, synaptic connectivity and/or memory capacity of neuronal networks in a subject as described herein, or in a method of preventing or treating pathological stress or at least one symptom thereof in a subject in need thereof, without exposing the nanoparticles or nanoparticle aggregates administered to said subject to an electric field and preferably without exposing it to any other external activation source, such as a light source, a magnetic field or an ultrasound source. Also described herein is a kit as described herein for preventing or treating pathological stress or at least one symptom thereof in a subject without exposing the nanoparticles or nanoparticle aggregates administered to the subject to an electric field, and preferably without exposing it to any other external activation source, such as a light source, a magnetic field, or an ultrasound source.

In a particular aspect, the nanoparticles or nanoparticle aggregates described herein are used in a method of treatment for enhancing physical performance or enhancing learning, memory, perception, attention and/or decision making in a subject, or for enhancing physical performance or enhancing learning, memory, perception, attention and/or decision making in a subject in need of such treatment, without exposing the nanoparticles or nanoparticle aggregates to an electric field, and preferably without exposing them to any other external activation source, such as a light source, a magnetic field or an ultrasound source.

In rodents, usually in mice, reliable evidence of psychometric intelligence can be obtained from a set of tests involving different tasks. These tests typically include learning tasks such as scent recognition or spatial navigation. The learning test is related to sensory, motor or motivational requirements imposed on the animal. For example, to assess reasoning for mice, tests based on The "quick mapping" concept (Carey S et al, Stanford Children Language Conference (Proceedings of The Standard Language Conference), 1978, 15, 17-29: obtaining a single new word (acquring a singlenew word)), to assess The attention task of mice, The "mouse Stroop test" can be used, and to assess The efficacy of The working memory or working memory capacity of mice, The "radial arm maze" test (Matzel L.D et al, human Current directives in Psychological Science, 201342, 22(5), The framework of intelligence 348: a summary from human and animal studies (The architecture of understanding. Convergining observations of humans and animals) can be used.

IQ testing can be used to assess the memory of humans. IQ tests such as Raven's Matrix or the Wechsler Adult mental Scale (Wechsler Adult Intelligent Scale) are well known to the skilled artisan and are commonly used to assess working memory in humans. The Stroop Color Word Interference Test (Stroop Color-Word Interference Test) (Stroop JR, Journal of Experimental psychological, 1935, 18, 643-.

In another particular aspect, the nanoparticles or nanoparticle aggregates described herein are used to enhance nerve/neuron connections, functional connectivity and/or synaptic plasticity in the brain of a subject in need of such treatment without exposing the nanoparticles or nanoparticle aggregates to an electric field, and preferably without exposing them to any other external activation source, such as a light source, a magnetic field or an ultrasound source.

In a typical aspect, the nanoparticles or nanoparticle aggregates described herein are used for the prevention or treatment of a subject suffering from a change in brain functional activity without exposing the nanoparticles or nanoparticle aggregates to an electric field, and preferably without exposing them to any other external activation source, such as a light source, a magnetic field or an ultrasound source.

In another particular aspect, the nanoparticles or nanoparticle aggregates described herein are used for preventing or treating a subject suffering from pathological stress or at least one symptom thereof, in particular from chronic stress, without exposing said nanoparticles or nanoparticle aggregates to an electric field, and preferably without exposing them to any other external activation source, such as a light source, a magnetic field or an ultrasound source. All living organisms strive towards homeostasis, which is called homeostasis. This balance is threatened by certain physical and psychological events. The interface between the incoming sensory information and the assessment process is formed by the limbic brain structures, including the hippocampus, amygdala, and prefrontal cortex. Various conditions may trigger stress, such as novelty, uncertainty, frustration, conflict, fear, pain, and the like. Stress can also be caused by continued exposure to adverse environments involving stimuli such as noise, pollution, and interpersonal conflicts.

The pathological stress resulting from such cumulative and/or repetitive events alters the structure (morphology) and/or connectivity of brain cells, and/or the functional properties of brain cells. Thus, pathological stress seriously affects health and limits the quality of life of humans.

Uncontrolled stress can have serious adverse consequences and cause symptoms, including reduced learning and memory. At mild stress levels, certain neurochemical systems (e.g., catecholamines, glucocorticoids) may affect learning. As the level of stress increases (duration and/or intensity), several temporary and permanent changes are observed in the hippocampus, including altered synaptic plasticity, changes in cell morphology, inhibition of adult neurogenesis, and/or neuronal destruction or atrophy (these changes are described herein as symptoms of pathological stress). These stress-related changes in the brain affect the learning and memory processes. Indeed, the hippocampus, amygdala and prefrontal cortex undergo stress-induced structural remodeling that alters behavioral and physiological responses. Chronic stress induces atrophy of neurons in the hippocampus and prefrontal cortex, as well as in brain regions involved in memory, selective attention, and executive function, and leads to hypertrophy of neurons in the amygdala, a region of the brain involved in fear and aggression. Chronic stress can compromise, typically by reducing the ability to learn, memory and make decisions, and can be accompanied by an increase in aggressiveness.

The extensive observations from in vitro and in vivo electrophysiological studies have in agreement suggested that stress and stress hormones impair Long-Term Potentiation (LTP) (i.e., permanently promote neurotransmission at the synapse after natural or artificial stimulation of the synapse, a cellular mechanism believed to be plasticity in the brain and particularly involved in learning and memory).

There are many agents, such as hypnotics, anxiolytics, and beta blockers, that combat some of the problems associated with pathological excessive stress. Likewise, drugs that reduce oxidative stress or inflammation, block cholesterol synthesis or absorption, and treat insulin resistance or chronic pain may help to address the metabolic and neurological consequences of "pathological overstress". All of these drugs are valuable to some extent, but unfortunately each has its side effects and limitations (Kim J. et al Nature reviews Neuroscience, 2002, 3, 453: 462 hippocampus under stress, synaptic plasticity and memory loss (stressed hippoplasms, synthetic plasticity and location networks); McEwenB. X. physiologial Review, 2007, 87, 873: 904: Physiology and neurobiology of stress and adaptation: central effects of the brain). The nanoparticles described herein can now advantageously be used for the treatment of subjects suffering from such pathological stress, in particular from chronic stress, typically subjects in the brain where stress-related changes as described above have been detected.

The term "treatment" refers to a therapeutic treatment or measure capable of preventing, alleviating or curing pathological stress or symptoms thereof, particularly chronic stress, as described above. Such treatment is intended for use in a mammalian subject, preferably a human subject in need thereof. Contemplated for this is a subject that has been identified (diagnosed) as having, or is considered to be "at risk" for developing, a pathological stress as described herein, for which treatment is prophylactic or preventative treatment. A particular subject suffering from pathological stress is a subject that has been prescribed a drug selected from the group consisting of hypnotics, anxiolytics, and beta blockers.

Pathological stress is different from oxidative stress. According to M.Auffan et al (see M.Auffan et al, Environmental Pollution 157(2009) 1127-1133: Chemical Stability of metal nanoparticles: parameters for controlling their potential cytotoxicity in vitro (Chemical Stability of metallic nanoparticles: a parametric controlling the same cellular toxicity in vitro)), oxidative stress is a state of redox imbalance in which ROS (reactive oxygen species) produced (by the cell or nanoparticle itself) overwhelms the antioxidant defense of the cell, causing undesirable biological consequences: macromolecular, lipid, DNA or protein damage leads to excessive cell proliferation, apoptosis, lipid peroxidation or mutagenesis. In cells, ROS production can be tracked, for example, by fluorescent dyes such as dichlorofluorescein diacetate.

The following examples and their corresponding figures illustrate the invention without limiting its scope.

Drawings

Figure 1. two simplified burst schemes outline some parameters that can be extracted from the electrical activity record. Parameters describing the general activity (spike), burst (burst), burst interval (IBI) and burst period) and burst structure (burst duration, burst plateau, burst amplitude, burst peak potential interval (ISI) and burst area) are indicated. The Standard Deviation (SD) of these parameters is a measure of the regularity of the general activity and burst structure, respectively. Coefficient of variation with time (CV)_{Time of day}) Reflecting the temporal regularity of the activity pattern of each unit. CV of_{Time of day}Calculated from the ratio of the standard deviation and the mean of the parameters. Coefficient of Variation (CV) between networks_Network) Reflecting the synchronization between neurons within the network. CV of_NetworkCalculated by the ratio of the standard deviation to the mean of the parameters on the network. Large CV of_NetworkValues imply a wide range of activity variation across the network, meaning low synchronization and high synaptic plasticity and synaptic connectivity.

Figure 2 functional effects of nanoparticles from example 1 on frontal cortex network activity observed in the "nanoparticles" group when compared to water used in the "control" group. The results indicate that synaptic plasticity and synaptic connectivity is higher at the cellular level in the presence of nanoparticles.

Figure 3 functional effect of nanoparticles from example 2 on frontal cortex network activity observed in the "nanoparticles" group when compared to water used in the "control" group. The results indicate that synaptic plasticity and synaptic connectivity is higher at the cellular level in the presence of nanoparticles.

Figure 4 functional effects of nanoparticles from example 3 on frontal cortex network activity observed in the "nanoparticles" group when compared to water used in the "control" group. The results indicate that synaptic plasticity and synaptic connectivity is higher at the cellular level in the presence of nanoparticles.

Figure 5 functional effects of nanoparticles from example 5 on frontal cortex network activity observed in the "nanoparticles" group when compared to water used in the "control" group. The results indicate that synaptic plasticity and synaptic connectivity is higher at the cellular level in the presence of nanoparticles.

Examples

In vitro study of neurons

At the neuronal level, the patch-clamp technique is very useful for detecting action potentials, since it allows direct measurement and control of the membrane potential of neurons at the same time.

This technique was used to evaluate the effect of nanoparticles on individual neurons.

In vitro study of neuronal networks

Distributed neuronal cultures coupled with a multi-electrode array (MEA) are widely used to better understand the complexity of brain networks. In addition, the use of discrete neuron assemblies allows manipulation and control of the connectivity of the network (Poli D. et al, Frontiers in Neural Circuits, 2015, 9 (paper 57), 1-14: Functional connectivity in in vitro neuron assemblies).

The MEA system enables non-invasive, persistent, simultaneous extracellular recordings from multiple sites in a neuronal network in real time, increasing spatial resolution, thereby providing a robust measure of network activity. The simultaneous collection of action potential and field potential data over a long period of time allows monitoring of network functions resulting from the interaction of all cellular mechanisms responsible for spatio-temporal pattern generation (Johnstone A.F.M. et al, neurobiology (2010), 31: 331-^stcentury)). In contrast to patch clamp and other single electrode recording techniques, MEA measures the response of the entire network, integrating the overall information of all receptor, synapse and neuron type interactions present in the network (Novellino a et al, Frontiers in Neuroengineering. (2011), 4(4), 1-14, developing microelectrode array-based neurotoxicity assaysTest: laboratory-to-laboratory reproducibility was assessed using neuroactive chemicals (Development of micro-electrode array based tests for neuroactive chemicals). Thus, MEA recordings have been used to understand neuronal communication, information coding, dissemination and processing in neuronal cultures (Taketani, M et al (2006) & Advances in network electrophysiology, New York, NY: Springer; Obien et al, front in Neurosciences, 2015, 8 (423): Revealing neuronal function by microelectrode array recordings). MEA technology is a complex phenotypic high-content screen for characterizing functional changes in network activity in electroactive cell cultures and is very sensitive to neurogenesis as well as neural regeneration and neurodegeneration. In addition, neuronal networks grown on MEA are known to respond to neuroactive or neurotoxic compounds in approximately the same concentration range that alters intact mammalian nervous system function (Xia et al, Alcohol, 2003, 30, 167: tissue-type electrophysiological responses of cultured neuronal networks to ethanol (Histidistribution of cultured neuronal networks to ethanol); Gramowski et al, European Journal of neurosciences, 2006, 24, 455: Functional screening of traditional antidepressants (Functional cortical neuronal networks grown on multielectrode neurochips) with primary cortical neuronal networks, biological assays with primary cortical neuronal networks with GABA therapeutic neuronal networks, cortical neuronal networks, biological networks, and cortical stimulation by stimulation of cortical neuronal networks (cortical stimulation of cortical neuronal activity in vitro) by alternating stimulation of cortical neuronal activity in vitro by stimulation of cortical neuronal activity in the range of 150MHz and cortical stimulation by modulation of cortical activity of cortical neuronal activity in the range of cortical stimulation of cortical activity in the range of cortical neuronal activity in the range of cortical stimulation of 10 MHz, stimulating cortical activity in vitro by modulation of cortical stimulation of cortical activity in the range of cortical neuronal networks grown on multielectrode electromagnetic field with 150MHz carrier wave pulsed with an alternating10 and 16 Hzmodulation)).

This technique was used to evaluate the effect of nanoparticles on neuronal networks.

In vivo study of neuronal networks

The effect of the nanoparticles of the invention on the neuronal network of an animal is evaluated taking into account an appropriate animal model.

For example, the maze is used to study spatial learning and memory in rats or mice. Research using the maze helps to reveal general principles about learning that are applicable to many species, including humans. The maze is now commonly used to determine whether different treatments or conditions affect the learning and memory of rats.