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Interfacing Graphene-Based Materials With Neural Cells

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Interfacing Graphene-Based Materials With Neural Cells
Published on October 8, 2021
Written by National Institutes for Health

Interfacing Graphene-Based Materials With Neural Cells Gene-editing-The-McGill-Tribune

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.
Graphene (G) is a single- or few-layered sheet of Sp2-bonded carbon atoms tightly packed in a two-dimensional (2D) honeycomb lattice, with a thickness of only 0.34 nm (Geim, 2009). Each carbon atom has three μ-bonds and an out-of-plane π-bond that can bind with neighboring atoms (Geim, 2009), making G the thinnest compound ever known at one atom thick and the strongest compound discovered.
Moreover, it is light, flexible and transparent and both electrically and thermally highly conductive, which opens the possibility of using it in a broad spectrum of applications, including supercapacitors (Hess et al., 2011; Sahoo et al., 2015; Casaluci et al., 2016), flexible electronics (Eda et al., 2008; Meric et al., 2008), printable inks (Zhu et al., 2015; Bonaccorso et al., 2016), batteries (Hassoun et al., 2014; Dufficy et al., 2015), optical and electrochemical sensors (Pumera, 2009; Du et al., 2010; Kang et al., 2010), energy storage (El-Kady and Kaner, 2013; Bonaccorso et al., 2015; Ambrosi and Pumera, 2016) and medicine (Novoselov et al., 2012; Casaluci et al., 2016; Kostarelos et al., 2017; Reina et al., 2017).
In the last few years, biomedical applications of G have attracted an ever-increasing interest, including the use of G and GRMs for bioelectrodes, bioimaging, drug/gene/peptide delivery, nanopore-based DNA-sequencing, stem cell differentiation and tissue engineering (Feng et al., 2013; Yang et al., 2013). Moreover, GRMs have generated great interests for the design of nanocarriers and nanoimaging tools, two- and three-dimensional tissue scaffolds, anti-bacterial coatings and biosensors (Bitounis et al., 2013; Ding et al., 2015).
The interest in using GRMs in medicine lies chiefly upon the extraordinary properties of G, including its mechanical properties, flexibility, transparency, thermo-electrical conductivity and good biocompatibility. GRMs could therefore overcome the limitations of metals and silicon, which are currently used for implantable devices, but are characterized by elevated stiffness, high inflammatory potential and poor long-term stability in physiological environments.
Moreover, the biomedical field witnesses a strong need for innovative therapies to assess the increasing demand of more specific, safer and effective treatments for pathological conditions. Given these premises, a large amount of research on G focuses on medical applications, and particularly in the field of neurology, where its mechanical and electronic features make it a strong candidate for replacing current devices (Kostarelos et al., 2017; Reina et al., 2017).
The biomedical applications of G represent a field in continuous expansion. Traditional treatments for central nervous system (CNS) disorders present a number of challenges, thus, developing new tools that outperform the state of the art technologies for imaging, drug delivery, neuronal regeneration and electrical recording and sensing is one of the main goal of modern medicine and neuroscience (Baldrighi et al., 2016).
Since the development of carbon-related materials, nanotechnology has strongly impacted a number of applications (Figure ​(Figure1)1) including: drug, gene and protein delivery, to cross the blood-brain barrier (BBB) and reach compromised brain areas; neuro-regenerative techniques to restore cell-cell communication upon damage by interfacing two (2D) or three (3D) dimensional scaffolds with neural cells; highly specific and reliable diagnostic tools, for in vivo sensing of disease biomarkers by cell labeling and real-time monitoring of biological active molecules; and neuronal activity monitoring and modulation, by highly sensitive electrodes for recordings and G-based platforms for electrical local stimulation (Mattei and Rehman, 2014; John et al., 2015; Chen et al., 2017; Kostarelos et al., 2017; Reina et al., 2017).
The key goal of any drug delivery system is to create a smart tool that recognizes specific targets and releases the drug in a controlled way (Allen and Cullis, 2004). The main limitation of G-based applications in neuroscience is its very low accumulation in the brain parenchyma upon intravenous injection. Once injected intravenously, G will engage with ions, lipids and proteins, resulting in the aggregation of the material and formation of a biomolecular corona that might affect the distribution of G and trigger inflammatory responses (Dell’Orco et al., 2010).
In addition, nanosheets can be phagocytosed by macrophages, inducing activation and release of pro-inflammatory cytokines (Zhou et al., 2012), and interact with several blood components inducing hemolysis (Liao et al., 2011). Last but not least, G nanosheets could accumulate in the reticulo-endothelial system rather than in the tissue to which they are targeted (McCallion et al., 2016).
One of the main applications of G-based drug delivery systems is anticancer therapy, by linking G composites with chemotherapeutics. Given their strong optical absorbance in the NIR region, G-based hybrid materials are also intensively studied for their promising applications in cancer phototherapy (Liu et al., 2011; Robinson et al., 2011; Yang et al., 2012; Hönigsmann, 2013). The rationale beyond this approach is to exploit the heat produced by the G accumulated in tumor regions upon NIR laser stimulation to kill cancer cells.
This technique was successfully applied in vitro using U251 glioma cells (Markovic et al., 2011). Such experimental approaches are of special interest, as they might help overcoming the limitations imposed by the BBB (Abbott, 2013), and are very promising especially for the treatment of very resistant and aggressive tumors, such as the glioblastoma.
Similarly to drug delivery, also genetic engineering can exploit G properties and open new opportunities in biomedicine. The concept in this case would be to deliver nucleic acids, i.e., DNA or various types of RNA molecules, including miRNA and shRNA, to specific target cell populations, to restore physiological conditions (Cheng et al., 2016).
The development of non-viral systems is of great importance for future medical approaches as G could allow overcoming some of the intrinsic limitations of viral systems, such as difficulties in accommodating long nucleic acids, batch-to-batch variations, elevated costs and the immunogenicity of viral vector systems (Kim et al., 2011; John et al., 2015).
Different strategies have been developed, including the decoration with positively charged polymers (PEI, BPEI), dendrimers (PAMAM) and polysaccharides, which enhance gene transfection efficiency by promoting the interaction with the cell membrane (Liu et al., 2014; Paul et al., 2014). Being the technique of functionalization the same, both drugs and genes can be delivered simultaneously using G-based hybrid materials (Zhang L. et al., 2011).
This would exhibit a synergic effect, as it would bring a significant enhancement of drug as well as transfection efficiency. On this line, G-nanosheets were functionalized with the cationic polymer PEI, a non-viral gene vector that forms strong electrostatic interactions with the negatively charged phosphate groups of both RNA and DNA (Feng et al., 2011). A step further was taken by Chen et al. (2011) that used PEI-functionalized GO for gene delivery yielding a high transfection efficiency in the absence of any cytotoxic effect.
In summary, G-based delivery systems, when conveniently functionalized or associated with complementary technologies, represent promising candidates for both diagnostics (i.e., imaging) and therapeutics (i.e., drug and gene delivery) neuroscience applications.
Moreover, in spite of few studies showing toxic effects of exposure the nervous system to bare G and rGO (Bramini et al., 2016; Mendonça et al., 2016b; Rauti et al., 2016), to date there is no solid evidence that functionalized-G is harmful to neuronal cells and the BBB.
Since G-based technologies for biomedical applications are constantly and rapidly evolving, the near future may see the development of new safe and highly neurocompatible materials.
In the past few years, GRMs have been studied and used in a wide range of technological fields, including biomedical applications. The treatment of neurological disorders through non-invasive pharmacological approaches is still a major challenge. It is crucial for scientist to develop strategies for efficient cargo delivery of drugs or biomolecules or even genes to the brain, bypassing the BBB while preserving its structure and vital functions.
One of the purposes of nanomedicine is indeed to create innovative ways for cell-targeting and drug-controlled release by avoiding surgery or other approaches that are very invasive for the patient. In this scenario, the choice of the appropriate ligand-receptor complex is a key design element when constructing nano-carriers, as well as the choice of the material, the size and eventual functionalization.
While receptor-mediated transcytosis is a fundamental pathway for BBB crossing, the development of next-generation nano-carriers, like 2D-materials, and the investigation and optimization of alternative routes for delivery, such as intranasal administration, is of utmost importance for the scientific community.
Besides the “BBB challenge”, other aspects of neuroscience could benefit of the latest developments in graphene research. Neuro-oncology may profit from the development of G nanosheets and G NPs for tumor-targeted imaging, photothermal therapy, and anticancer drug delivery and gene therapy.
New electrical, chemical and optical sensors may have great impact for neuro-intensive care and neuro-monitoring. Moreover, the combination of different forms and states of G, diverse chemical functionalization and the possible association with other biomaterials to form G-based composites, may allow to devise an all-in one tool for both diagnosis and therapy, thus effectively building a powerful theranostic device.
Finally, tissue-engineering research is expected to develop novel brain-implant interfaces based on G, to exploit the material electrical conductivity and enhance cell-cell communication and repair. Besides the experimental and clinical evidence, MD studies are emerging as an important aspect of material research, as they provide extremely precise indications and predictions on G/cell and G/protein interactions, guiding the researcher to design more powerful G-based devices.
Nevertheless, despite initial studies demonstrated the biocompatibility of G, especially when conjugated with other materials in 2D and 3D scaffolds, only few systems were demonstrated to be successful in vivo. Further investigations are still required, in particular about the biological effects of long-term treatment with G materials, before the promised technological applications can be fully exploited in and beyond neuroscience.
This is taken from a very long article. Read the rest here: ncbi.nlm.nih.gov
Header image: The McGill Tribune

Thanks to: https://principia-scientific.com


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