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The safety of graphene in human health: what science says about it

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The safety of graphene in human health:

what science says about it

Part II. Are graphene materials safe for humans?

The family of graphene materials comprises a wide range of two-dimensional (2D) carbon nanostructures in the form of sheets that differ from each other by the particularities derived from the production method or by the innumerable functionalizations that can be performed after its obtaining. In 2022, Nature magazine, one of the most important scientific journals in the world, published a study in which 36 products from graphene suppliers from countries such as the United States, Norway, Italy, Canada, India, China, Malaysia and England were analyzed, concluding that graphenes represent a heterogeneous class of materials with variable characteristics and properties, whether mechanical, thermal, electrical, optical, biological, etc., which can be transferred to a large number of three-dimensional (3D) compounds to modify or create new products.

“Undoubtedly, graphene and nanotechnology in general continue to be controversial issues as they confront us with a world that is difficult to see and understand, but with simply amazing effects”

Are graphene materials safe?

Graphene materials promise to be an important tool within biomedical technologies. In principle, its benefits can be used for the design of diagnostic elements such as sensors and devices for images up to neural interfaces, gene therapy, drug delivery, tissue engineering, infection control, phototherapy for cancer treatment, bioelectronic and dental medicine, among other. But for them to be truly used in this type of technology, their interactions with the biological environment must first be understood or, failing that, ensure that their presence does not alter the natural environment of the cells. In this sense, numerous studies have been carried out with the different forms, presentations, and available concentrations of graphene whose findings have gradually paved the way for its safe use in biomedical technologies:

i) Graphene materials in their free form. In in vitro tests, exposure of human lung epithelial cells to graphene sheets at concentrations lower than 0.005 mg/ml did not cause significant changes in their morphology or adhesion,2,3 nor was cytotoxic activity identified in stem cells derived from adipose tissue. human, periodontal ligament and dental pulp exposed to 0.5 mg/ml of GO,4 even and understanding a possible dose-size dependent effect, other investigations report safe concentrations below 40 mg/ml or, that do not exceed 1, 5% w/v. 5-8

Finally, one of the most recent in vivo studies published by the University of Manchester, United Kingdom, on the pulmonary response of mice exposed to graphene oxide (GO) in the respiratory tract, did not identify significant damage or pulmonary fibrosis at 90-day follow-up. These results provide solid grounds for the safety of these nanostructures without underestimating basic safety measures, such as avoiding their inhalation.9 Likewise, scientists from the University of Trieste, Italy, analyzed the impact of graphene materials on the skin, reporting low toxicity on cells.10

“It is unlikely that graphene materials in their free form are used to be in contact with the biological environment, they are generally functionalized or immobilized in other materials to develop an application”

ii) Functionalized graphene materials. Functionalization is the term that refers to the chemical modification of a nanomaterial to give it a “function”, that is, to facilitate its incorporation with other compounds or to benefit its biocompatibility and better direct its use by anchoring functional groups, molecules, or nanoparticles. A study published in the journal Nature Communications on graphene bioapplications highlights the importance of its functionalization with amino groups to make it more compatible with human immune cells.11,12

“The most common functionalization of graphene is the anchoring of oxygenated groups on its surface, this material is known as graphene oxide”

iii) Immobilization in polymers. The use of graphene materials as nano-filling for plastics, resins, coatings, etc., is the most common way in which these nanostructures are used. For the biomedical sector, its immobilization in polymers has shown good biocompatibility and stimulation of cell proliferation; antimicrobial activity and improvement of the mechanical properties of polymers, being classified as excellent candidates for the manufacture of bone fixation devices, molecular scaffolds, orthopedic implants, or dental materials.13-15

Given the great potential of graphene materials in health sciences, but also due to the many questions about their safety, an international research team from the European Graphene Flagship project, led by EMPA (German acronym for the Federal Institute for Testing and Materials Research), conducted a study to assess the potential health effects of graphene materials immobilized within a polymer; the results showed that the graphene particles released from said polymeric compounds after abrasion induce insignificant effects.16

“It is reassuring to see that this study shows negligible effects, confirming the viability of graphene for mass applications. Andrea C. Ferrari, Graphene Flagship Science and Technology Officer.” 17,18

Energeia-Graphenemex, the pioneering Mexican company in Latin America in the research and development of applications with graphene materials, throughout its 10-year career has overcome numerous scientific and industrial challenges to reach the market with products for industrial use. In 2018, it began to explore the antimicrobial capabilities of its products with excellent results in vitro and in a relevant environment; currently, and in conjunction with other research centers, it is carrying out evaluations to explore the potential of its materials as nano-reinforcement of biopolymers.

Drafting: EF/DHS

References

  1. Cytotoxicity survey of commercial graphene materials from worldwide. npj 2D Materials and Applications (2022) 6:65
  2. Biocompatibility of Pristine Graphene Monolayers, Nanosheets and Thin Films. 2014, 1406.2497.
  3. Preliminary In Vitro Cytotoxicity, Mutagenicity and Antitumoral Activity Evaluation of Graphene Flake and Aqueous Graphene Paste. Life 2022, 12, 242
  4. Biological and physico-mechanical properties of poly (methyl methacrylate) enriched with graphene oxide as a potential biomaterial. J Oral Res 2021; 10(2):1
  5.  Graphene substrates promote adherence of human osteoblasts and mesenchymal stromal cells. Carbon. 2010; 48: 4323–9
  6. Multi-layer Graphene oxide in human keratinocytes: time-dependent cytotoxicity. Prolifer Gene Express Coat 2021; 11:1
  7. Cytotoxicity assessment of graphene-based nanomaterials on human dental follicle stem cells. Colloids Surf B Biointerfaces. 2015; 136:791
  8. Arabinoxylan/graphene-oxide/nHAp-NPs/PVA bionano composite scaffolds for fractured bone healing. 2021. J. Tissue Eng. Regen. Med. 15, 322.
  9. Size-Dependent Pulmonary Impact of Thin Graphene Oxide Sheets in Mice: Toward Safe-by-Design. Adv. Sci. 2020, 7, 1903200
  10. Differential cytotoxic effects of graphene and graphene oxide on skin keratinocytes. 2017. Sci Rep 7, 40572
  11. Amine-Modified Graphene: Thrombo-Protective Safer Alternative to Graphene Oxide for Biomedical Applications. ACS Nano 2012, 6, 2731
  12. Single-cell mass cytometry and transcriptome profiling reveal the impact of graphene on human immune cells. Nature Communications, 2017, 8: 1109,
  13. In-vitro cytotoxicity of zinc oxide, graphene oxide, and calcium carbonate nano particulates reinforced high-density polyethylene composite. J. Mater Res. Technol. 2022. 18: 921
  14. Graphene-Doped Polymethyl Methacrylate (PMMA) as a New Restorative Material in Implant-Prosthetics: In Vitro Analysis of Resistance to Mechanical FatigueJ. Clin. Med. 2023, 12, 1269
  15. High performance of polysulfone/ Graphene oxide- silver nanocomposites with excellent antibacterial capability for medical applications. Matter today commun. 2021. 27
  16. Hazard assessment of abraded thermoplastic composites reinforced with reduced graphene oxide. J. Hazard Mater. 2022. 435. 129053
  17. https://www.empa.ch/web/s604/graphene-dust
  18. https://www.graphene-info.com/researchers-asses-health-hazards-graphene-enhanced-composites

Graphene: The next revolution in biomedical applications

Posted on | by Energeia Graphenemex

Graphene:

The next revolution in biomedical applications

Part I. Tissue Engineering

Advances in medicine have reached levels unimagined until recently. Among them, tissue engineering has an important participation. With it is possible to combine cells, biomaterials and biologically active molecules with the aim of repairing or replicating tissues or organs with a function similar to that of the original structure. In principle, biomaterials are used as molecular scaffolds to act as a three-dimensional (3D) support or guide for the anchoring and growth of the cells that will be in charge of forming the new tissue.

The first molecular scaffolds were designed with natural materials such as collagen, glycosaminoglycans (GAGs), chitosan, and alginates; then with artificial compounds such as polylactic acid (PLA), polyglycolic acid (PGA), poly(lactic-co-glycolic) acid (PLGA), polyurethanes (PUs), polytetrafluoroethylene (PTFE), polyethyleneterephthalate (PET); bioceramics such as hydroxyapatite (HA) and tricalcium phosphate; metals such as stainless steel, chrome-cobalt alloys (Co-Cr) or titanium alloys (Ti) and recently, new research is oriented towards the use of nanotechnology.

The relationship between nanotechnology and tissue engineering is due to the fact, that the extracellular matrix (ECM) that helps cells unite and communicate with each other, is made up of a network of nanometer-sized fibers made up of bioactive molecules. It is at this point where nanotechnology opens new possibilities for regenerative medicine, since it has been proven that the use of materials that act on the same nanometric scale as the ECM favors mimicking the physiological environment of the organism to stimulate cell growth and differentiation in a more natural environment.

Among the most studied nanomaterials in recent years are graphene materials, which consist of nanometric sheets of carbon atoms organized in two-dimensional (2D) hexagonal networks. Among the most interesting properties for tissue engineering are: its large surface area, mechanical resistance, thermal conductivity, biocompatibility and finally, an extraordinary ability to share its properties with other materials to improve their original characteristics.

For example, the use of graphene materials within the 3D architecture of certain biopolymers in tests carried out on heart, liver, bone, cartilage, and skin tissues has shown substantial improvements in their physicochemical, mechanical, electrical and biological properties, achieving excellent response. for stem cell adhesion and differentiation.

In 2022, the Andaltec technology center (Spain) reported the development of a material from polymers derived from graphene by 3D printing with great potential for the regeneration of muscle tissue. They demonstrated that in the presence of graphene derivatives, cells contract and expand without an external stimulus, therefore, it has great potential for use in regenerative medicine.

On the other hand, the Division of Postgraduate Studies and Research (DEPeI) on Odontology, UNAM and the National School of Higher Studies (ENES) León Unit, Mx., through a study published in J Oral Res 2021 supports the possibilities of graphene oxide (GO) in the design of biomaterials for dental use. The results of the research carried out with Graphenemex® GO samples, concluded that this nanomaterial in combination with polymethylmethacrylate (PMMA), in addition to improving its physical-mechanical properties, also demonstrated good compatibility and an interesting stimulation of cell proliferation when being evaluated on cultures with gingival-fibroblasts, dental-pulp-cells and human osteoblasts.

In 2020, researchers from the University of Malaga (Spain) published another study that identified GO as the ideal material for regenerative medicine. The study carried out on an animal model, showed high biocompatibility of different types of graphene oxide with dopaminergic cells, favoring their maturation and protecting them from the toxic conditions of Parkinson’s disease. These results postulate GO as an adequate scaffold to test new drugs or develop constructs for cell replacement therapy of Parkinson’s disease.

Despite the large amount of research on the interactions of graphene materials with biological media, there is still a long way to go to have these biomaterials available and in clinical operation. Energeia- Graphenemex, the pioneering Mexican company in Latin America in the research and development of applications with graphene materials, in collaboration with other companies and research centers, seeks to contribute with science to understand these interactions in a security framework, to lay solid foundations on the use of graphene nanotechnology in the biomedical sector for the benefit of society.

Drafting: EF/DHS

References

  1. Graphene and its derivatives: understanding the main chemical and medicinal chemistry roles for biomedical applications. J Nanostructure Chem, 2022, 12:693
  2. Biological and physico-mechanical properties of poly (methyl methacrylate) enriched with graphene oxide as a potential biomaterial. J Oral Res 2021; 10(2):1
  3. Graphene-Based Antimicrobial Biomedical Surfaces. ChemPhysChem 2021, 22, 250
  4. Functionalized Graphene Nanoparticles Induce Human Mesenchymal Stem Cells to Express Distinct Extracellular Matrix Proteins Mediating Osteogenesis. Int J Nanomed 2020:15 2501
  5. Graphene Oxide and Reduced Derivatives, as Powder or Film Scaffolds, Differentially Promote Dopaminergic Neuron Differentiation and Survival. Front. Neurosci., 21 December 2020. Sec. Neuropharmacology Volume 14
  6. International Journal of Nanomedicine 2019:14 5753
  7. Biocompatibility Considerations in the Design of Graphene Biomedical Materials. Adv. Mat. Interfaces 2019, 6, 1900229
  8. Graphene based scaffolds on bone tissue engineering. Bioengineered, 2018, 9:1, 38
  9. When stem cells meet graphene: Opportunities and challenges in regenerative medicine. Biomaterials, 2018, 155, 236
  10. Graphene-based materials for tissue engineering. Adv. Drug Deliv. Rev. 2016,105, 255

Chapter 92 e: Tissue Engineering, Anthony Atala. 2023 McGraw Hill.

Graphene oxide: a promising alternative in nanotechnology

Posted on | by Energeia Graphenemex

Graphene oxide:

a promising alternative in nanotechnology

Since graphene was isolated for the first time in 2004 by the Manchester group, this nanomaterial has proven to be the most revolutionary for the development of new applications at an industrial level.

Graphene has extraordinary electrical, optical, thermal properties and high mechanical resistance. The properties of graphene are attributed to its structure in the form of two-dimensional (2D) sheets, made up of hexagonally bonded carbon atoms and a thickness of one carbon atom.

Currently there are different methods of graphene production, these can be classified into two methods, according to their origin, the “bottom-up” method and the “top down” method. The “bottom-Up” method consists in the creation of graphene structures through building blocks (atoms, molecules), for example, by Chemical Vapor Deposition (CVD); and the “top down” method involves the production of graphene from the oxidation of graphite. Graphite is made up of sheets of graphene that are stacked on top of each other. The following diagram represents the process for obtaining graphene from the oxidation of graphite.

Schematic diagram of the process for obtaining GO, through the oxidation of graphite.

The graphite oxidation process begins with the addition of graphite in sulfuric acid (H2SO4), with constant mechanical stirring. Subsequently, potassium permanganate (KMnO4) is slowly added, producing a chemical reaction that allows the graphite (graphene sheets stacked on top of each other) to be chemically modified in its structure. When KMnO4 reacts with H2SO4, it forms manganese oxide VII (Mn2O7), which is a very selective oxidizing agent on double bond aromatic compounds, such as graphite. The oxidizing agent molecularly attacks the structure of each graphene sheet in the graphite, grafting oxygenated functional groups (with oxygen), such as epoxide groups (C-O-C) and hydroxyl groups (-OH), on each sheet, and carboxyl groups (-COOH, CO2H ) on the edges of each sheet, obtaining graphite oxide and graphene oxide (GO), see Figure 1.

Figure 1. Structure of graphene oxide

The incorporation of oxygenated functional groups allows a material such as graphite, which is highly hydrophobic (which repels water) and a good electrical conductor, to become graphite oxide and graphene oxide (GO), highly hydrophilic materials, that is, they mix and disperse easily with water (See Figure 2). GO is chemically similar to graphite oxide, but structurally differs in the arrangement and number of stacked sheets.

Figure 2. (a and b) Behavior of graphite and graphene oxide (GO) in water, c) GO sheet, Transmission Electron Microscopy (TEM).

The GO can be defined as a single exfoliated graphene sheet or stack of few sheets (3-4) that is functionalized with different oxygenated groups. Among its main characteristics is that it is hydrophilic, insulating and hygroscopic (absorbs moisture). On the other hand, graphene oxide sheets possess a large surface area and exhibit high mechanical strength and flexibility.

Applications

Graphene oxide has attracted great interest in various fields of science and technology, due to its remarkable mechanical, chemical, and thermal properties, among others. So numerous investigations began, to take advantage of the properties of graphene oxide.

In 2011, the first investigations of the use of GO as a precursor in the large-scale production of graphene emerged, for use as filler/reinforcement material/in polymeric matrices, such as high-density polyethylene (HDPE) and low-density polyethylene (HDPE). density (LDPE).

By 2014, GO was considered feasible for use as a flame retardant agent. Research is still ongoing to functionalize it with different polymeric materials.

In 2017, the first reports of the manufacture of GO-based membranes began, since it is impermeable to gases and liquids, showing its ability to filter small particles, organic molecules and even its use for seawater desalination.

In 2018, Energeia-Graphenemex started research on graphene oxide as a new additive for the production of anticorrosive and antimicrobial coatings. By 2019, studies of graphene oxide in coatings with antibacterial behavior increased, associated with the fact that GO is capable of penetrating the cell membrane of bacteria, producing oxidative stress and inhibiting their reproduction.

In particular, the functionalization of GO allows it to be applicable in biological systems, development of biosensors for the identification of specific molecules, drug delivery systems, among others.

Energeia – Graphenemex®, a leading Mexican company in Latin America in research and production of graphene materials for the development of industrial applications. It has extensive experience in the production of graphene oxide (GO) on a large scale, with different degrees of oxidation and high quality for use in different applications and industries. Currently, it uses graphene oxide in the production of concrete additives and anticorrosive and antimicrobial coatings that are marketed under the Graphenergy brand.

References

  1. M. Fang, K. Wang, H. Lu, Y. Yang y S. Nutt, «Covalent polymer functionalization of graphene nanosheets and mechanical properties of composites,» Journal of Materials Chemistry, vol. 19, pp. 7098-7105, 2009.
  2. B. Dittrich, K.-a. Wartig, R. Mülhaupt y B. Schartel, «Flame-Retardancy Properties of Intumescent Ammonium Poly(Phosphate) and Mineral Filler Magnesium Hydroxide in Combination with Graphene,» Polymers, vol. 6, pp. 2875-2895, 2014.
  3. Y.-j. Wan, L.-x. Gong, L.-c. Tang, L.-b. Wu y J.-x. Jiang, «Mechanical properties of epoxy composites filled with silane-functionalized graphene oxide,» COMPOSITES PART A, vol. 64, pp. 79-89, 2014.
  4. J. Wang, C. Xu, H. Hu, L. Wan, R. Chen, H. Zheng, F. Liu, M. Zhang, X. Shang y X. Wang, «Synthesis , mechanical , and barrier properties of LDPE / Graphene nanocomposites using vinyl triethoxysilane as a coupling agent,» J. Nanopart Res, vol. 13, pp. 869-878, 2011.

The rise of graphene: advances and developments in the last decade

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The rise of graphene:

advances and developments in the last decade

Graphene is the most revolutionary nanomaterial of the 21st century and is considered the basic pillar for carbon nanochemistry, that is, the main element of all organic compounds.

Its versatility derives from its structure in the form of two-dimensional (2D) sheets, made up of carbon atoms linked in a hexagonal manner, and its importance lies in the extraordinary properties attributed to it and that have been conceived as the solution to innumerable social, environmental, scientific, technological and of course, economic needs.

Graphene sheet. High Resolution Transmission Electron Microscopy.
Energeia Collection – Graphenemex

Graphene allows matter to be modified to design compounds with new or improved characteristics, since it transfers its properties to the materials to which it is incorporated. This has allowed it to be used in the development of applications that seek to potentiate these properties, as shown in the following image.

Evolution – Graphene was first isolated in 2004 by Russian researchers Andre Geim and Konstantin Novoselov from the University of Manchester; subsequently, and thanks to their experiments, in 2010 they were awarded the Nobel Prize in Physics, as it was considered one of the most important discoveries of the century.

So important was the finding that in 2013 the European Union (EU) granted a budget of one billion euros to create the Graphene Flagship, an ambitious project valid for ten years with the aim of linking academia with industry, not only to understand its properties theoretically, but to fully exploit its benefits in real applications or products.

From that moment on, the progress of the investigations was so fast, and the expectations were higher and higher that, in 2017, the first edition of the ISO/TS 80004-13:2017 standard emerged (Ratified by the Spanish Association for Standardization in October 2020) for the normalization and standardization of Nanotechnology in new materials, including Graphene.

In the same 2020, a group of 70 researchers who are members of the Graphene Flagship, published the first manual with more than 500 pages on countless types of Graphene. By 2021, around 50 “spin-offs” and “startups” with different visions were registered within the organization, making the possibility of having a greater number of applications with Graphene or graphene materials at more affordable costs a reality by 2022.

In 2021 again the EU through the Federal Institute for Materials Research and Testing (BAM) with the new ISO-G-SCoPe project, set the objective of standardizing the methods to transfer Graphene to the industry, this as a result of the non-existence of production and quality standards, while, through the Versailles Project on Advanced Materials and Standards, under the direction of BAM, it seeks to validate the processes in a global test to convert them into standards.

Energeia Graphenemex® is the pioneer Mexican company in Latin America focused on the research and production of graphene materials for the development of applications at an industrial level. Among its strengths is the creation of patented methods and processes for replicable and large-scale production that ensures the availability of the appropriate graphene materials in accordance with the requirements of the applications it develops, either for its own products or as a strategic ally of other companies interested in innovating and improving their products with these materials.