Clinical potential of plasma-functionalized graphene oxide ultrathin sheets for bone and blood vessel regeneration: Insights from cellular and animal models.

Graphene and graphene oxide (GO), due to their unique chemical and physical properties, possess biochemical characteristics that can trigger intercellular signals promoting tissue regeneration. Clinical applications of thin GO-derived sheets have inspired the development of various tissue regeneration and repair approaches. In this study, we demonstrate that ultrathin sheets of plasma-functionalized and reduced GO, with the oxygen content ranging from 3.2 % to 22 % and the nitrogen content from 0 % to 8.3 %, retain their essential mechanical and molecular integrity, and exhibit robust potential for regenerating bone tissue and blood vessels across multiple cellular and animal models. Initially, we observed the growth of blood vessels and bone tissue in vitro using these functionalized GO sheets on human adipose-derived mesenchymal stem cells and umbilical vein endothelial cells. Remarkably, our study indicates a 2.5-fold increase in mineralization and two-fold increase in tubule formation even in media lacking osteogenic and angiogenic supplements. Subsequently, we observed the initiation, conduction, and formation of bone and blood vessels in a rat tibial osteotomy model, evident from a marked 4-fold increase in the volume of low radio-opacity bone tissue and a significant elevation in connectivity density, all without the use of stem cells or growth factors. Finally, we validated these findings in a mouse critical-size calvarial defect model (33 % higher healing rate) and a rat skin lesion model (up to 2.5-fold increase in the number of blood vessels, and 35 % increase in blood vessels diameter). This study elucidates the pro-osteogenic and pro-angiogenic properties of both pristine and plasma-treated GO ultrathin films. These properties suggest their significant potential for clinical applications, and as valuable biomaterials for investigating fundamental aspects of bone and blood vessel regeneration.

Application of the Van Cittert Algorithm for Deconvolving Loss Features in X-ray Photoelectron Spectroscopy Spectra.

The convolution of two physical entities, denoted as f and g, delineates the manner in which one entity undergoes modification in response to the other. This transformative process is mathematically represented by the expression f ⨂ g, symbolizing the convolution of the two entities in a resultant function h. Frequently, it becomes imperative to comprehend the magnitude of the induced modifications. From the derived function h, a crucial step involves the separation of the two original signals, a process commonly referred to as deconvolution. Various techniques have been proposed to facilitate the calculation of the deconvolution, with one notable approach originating in 1931 by van Cittert. The algorithm, based on an iterative method, has been scrutinized over time, notably by Bracewell and, more recently, by Jansson. This work represents the current state-of-the-art, focusing specifically on the analysis of Auger spectra obtained through XPS. Emphasis is placed on delineating the procedural aspects of the analysis, and the algorithm utilized in the open-source software RxpsG is comprehensively described.

Strain-Induced Plasmon Confinement in Polycrystalline Graphene.

Terahertz spectroscopy is a perfect tool to investigate the electronic intraband conductivity of graphene, but a phenomenological model (Drude-Smith) is often needed to describe disorder. By studying the THz response of isotropically strained polycrystalline graphene and using a fully atomistic computational approach to fit the results, we demonstrate here the connection between the Drude-Smith parameters and the microscopic behavior. Importantly, we clearly show that the strain-induced changes in the conductivity originate mainly from the increased separation between the single-crystal grains, leading to enchanced localization of the plasmon excitations. Only at the lowest strain values explored, a behavior consistent with the deformation of the individual grains can instead be observed.

Characterization of Carbon Nanostructures by Photoelectron Spectroscopies.

Recently, the scientific community experienced two revolutionary events. The first was the synthesis of single-layer graphene, which boosted research in many different areas. The second was the advent of quantum technologies with the promise to become pervasive in several aspects of everyday life. In this respect, diamonds and nanodiamonds are among the most promising materials to develop quantum devices. Graphene and nanodiamonds can be coupled with other carbon nanostructures to enhance specific properties or be properly functionalized to tune their quantum response. This contribution briefly explores photoelectron spectroscopies and, in particular, X-ray photoelectron spectroscopy (XPS) and then turns to the present applications of this technique for characterizing carbon nanomaterials. XPS is a qualitative and quantitative chemical analysis technique. It is surface-sensitive due to its limited sampling depth, which confines the analysis only to the outer few top-layers of the material surface. This enables researchers to understand the surface composition of the sample and how the chemistry influences its interaction with the environment. Although the chemical analysis remains the main information provided by XPS, modern instruments couple this information with spatial resolution and mapping or with the possibility to analyze the material in operando conditions at nearly atmospheric pressures. Examples of the application of photoelectron spectroscopies to the characterization of carbon nanostructures will be reviewed to present the potentialities of these techniques.

Tuning Surface Properties via Plasma Treatments for the Improved Capture of MicroRNA Biomarkers.

Advanced materials could bring about fundamental improvements in the evolution of innovative analytical devices, i.e., biosensors or lab-on-a-chip devices, in particular in the context of liquid biopsies. Here, plasma deposition processes were tested for the introduction of primary amines on silicon surfaces by tuning the amounts and availability of amino-charged residues. Different binary (CH4/NH3) and ternary (CH4/NH3/H2 and CH4/NH3/N2) mixtures of gases were used as feeds for the plasma treatments. The obtained surfaces were fully characterized for their chemical and physical properties before their use as capture materials in a functional test. Synthetic and fluorescently conjugated microRNA-21 (miR-21) was selected as the target molecule. The capture of miR-21 increased linearly with the increase in amino nitrogen measured on surfaces. The surface showing the most promising performance was further analyzed in different conditions, i.e., varying pH and time of incubation, incubation with different microRNAs, and possible elution of captured microRNAs. The apparent pH range of primary amines present on the surfaces was around 3.5-4. Positively charged surfaces prepared via PE-CVD were, therefore, demonstrated as being suitable materials for the capture of microRNA biomarkers, paving the way for their inclusion in biomedical devices for the purification and analysis of circulating biomarkers.

Multimodal bioimaging using nanodiamond and gold hybrid nanoparticles.

Hybrid core-shell nanodiamond-gold nanoparticles were synthesized and characterized as a novel multifunctional material with tunable and tailored properties for multifunctional biomedical applications. The combination of nanostructured gold and nanodiamond properties afford new options for optical labeling, imaging, sensing, and drug delivery, as well as targeted treatment. ND@Au core-shell nanoparticles composed of nanodiamond (ND) core doped with Si vacancies (SiV) and Au shell were synthesized and characterized in terms of their biomedical applications. Several bioimaging modalities based on the combination of optical and spectroscopic properties of the hybrid nano-systems are demonstrated in cellular and developing zebrafish larvae models. The ND@Au nanoparticles exhibit isolated ND's Raman signal of sp3 bonded carbon, one-photon fluorescence of SiV with strong zero-phonon line at 740 nm, two-photon excited fluorescence of nanogold with short fluorescence lifetime and strong absorption of X-ray irradiation render them possible imaging agent for Raman mapping, Fluorescence imaging, two-photon Fluorescence Lifetime Imaging (TP-FLIM) and high-resolution hard-X-ray microscopy in biosystems. Potential combination of the imaging facilities with other theranostic functionalities is discussed.

Gold nanostructures: synthesis, properties, and neurological applications.

Recent advances in technology are expected to increase our current understanding of neuroscience. Nanotechnology and nanomaterials can alter and control neural functionality in both in vitro and in vivo experimental setups. The intersection between neuroscience and nanoscience may generate long-term neural interfaces adapted at the molecular level. Owing to their intrinsic physicochemical characteristics, gold nanostructures (GNSs) have received much attention in neuroscience, especially for combined diagnostic and therapeutic (theragnostic) purposes. GNSs have been successfully employed to stimulate and monitor neurophysiological signals. Hence, GNSs could provide a promising solution for the regeneration and recovery of neural tissue, novel neuroprotective strategies, and integrated implantable materials. This review covers the broad range of neurological applications of GNS-based materials to improve clinical diagnosis and therapy. Sub-topics include neurotoxicity, targeted delivery of therapeutics to the central nervous system (CNS), neurochemical sensing, neuromodulation, neuroimaging, neurotherapy, tissue engineering, and neural regeneration. It focuses on core concepts of GNSs in neurology, to circumvent the limitations and significant obstacles of innovative approaches in neurobiology and neurochemistry, including theragnostics. We will discuss recent advances in the use of GNSs to overcome current bottlenecks and tackle technical and conceptual challenges.

Multimodal Gold Nanostars as SERS Tags for Optically-Driven Doxorubicin Release Study in Cancer Cells.

Surface Enhanced Raman Scattering (SERS) active gold nanostars represent an opportunity in the field of bioimaging and drug delivery. The combination of gold surface chemical versatility with the possibility to tune the optical properties changing the nanoparticles shape constitutes a multimodal approach for the investigation of the behavior of these carriers inside living cells. In this work, SERS active star-shaped nanoparticles were functionalized with doxorubicin molecules and covered with immuno-mimetic thiolated polyethylene glycol (PEG). Doxorubicin-conjugate gold nanoparticles show an intense Raman enhancement, a good stability in physiological conditions, and a low cytotoxicity. The strong adsorption of the anticancer drug doxorubicin in close contact with the gold nanostars surface enables their use as SERS tag imaging probes in vivo. Upon laser irradiation of the nanoparticles, a strong SERS signal is generated by the doxorubicin molecules close to the nanostars surface, enabling the localization of the nanoparticles inside the cells. After long time irradiation, the SERS signal drops, indicating the thermally driven delivery of the drug inside the cell. Therefore, the combination of SERS and laser scanning confocal microscopy is a powerful technique for the real-time analysis of drug release in living cells.

Carbon Nanomaterials: Synthesis, Functionalization and Sensing Applications.

Recent advances in nanomaterial design and synthesis has resulted in robust sensing systems that display superior analytical performance. The use of nanomaterials within sensors has accelerated new routes and opportunities for the detection of analytes or target molecules. Among others, carbon-based sensors have reported biocompatibility, better sensitivity, better selectivity and lower limits of detection to reveal a wide range of organic and inorganic molecules. Carbon nanomaterials are among the most extensively studied materials because of their unique properties spanning from the high specific surface area, high carrier mobility, high electrical conductivity, flexibility, and optical transparency fostering their use in sensing applications. In this paper, a comprehensive review has been made to cover recent developments in the field of carbon-based nanomaterials for sensing applications. The review describes nanomaterials like fullerenes, carbon onions, carbon quantum dots, nanodiamonds, carbon nanotubes, and graphene. Synthesis of these nanostructures has been discussed along with their functionalization methods. The recent application of all these nanomaterials in sensing applications has been highlighted for the principal applicative field and the future prospects and possibilities have been outlined.

Highly Deformable, Ultrathin Large-Area Poly(methyl methacrylate) Films.

Poly(methyl methacrylate) (PMMA) is a glassy engineering polymer that finds extensive use in a number of applications. Over the past decade, thin films of PMMA were combined with graphene or other two-dimensional materials for applications in the area of nanotechnology. However, the effect of size upon the mechanical behavior of this thermoplastic polymer has not been fully examined. In this work, we adopted a homemade nanomechanical device to assess the yielding and fracture characteristics of freestanding, ultrathin (180-280 nm) PMMA films of a loaded area as large as 0.3 mm2. The measured values of Young's modulus and yield strength were found to be broadly similar to those measured in the bulk, but in contrast, all specimens exhibited a quite surprisingly high strain at failure (>20%). Detailed optical examination of the specimens during tensile loading showed clear evidence of craze development which however did not lead to premature fracture. This work may pave the way for the development of glassy thermoplastic films with high ductility at ambient temperatures.

Tuning the Oxygen Content of Reduced Graphene Oxide and Effects on Its Properties.

The need to recover the graphene properties in terms of electrical and thermal conductivity calls for the application of reduction processes leading to the removal of oxygen atoms from the graphene oxide sheet surface. The recombination of carbon-carbon double bonds causes a partial recovery of the original graphene properties mainly limited by the presence of residual oxygen atoms and lattice defects. However, the loss of polar oxygen-based functional groups renders the material dispersibility rather complicated. In addition, oxygen-containing functional groups are reaction sites useful to further bind active molecules to engineer the reduced graphene sheets. For these reasons, a variety of chemical processes are described in the literature to reduce the graphene oxide. However, it is greatly important to select a chemical process enabling a thin modulation of the residual oxygen content thus tuning the properties of the final product. In this work, we will present a chemical-processing technique based on the hydroiodic acid to carefully control the degree of residual oxidation. Graphene oxides were reduced using hydroiodic acid with concentrations from 0.06 to 0.95 mol L-1. Their properties were characterized in detail and tested, and the results showed that their oxygen content was finely tuned from 33.6 to 10.7 atom %. This allows carefully tailoring the material properties with respect to the desired application, which is exemplified by the variation of the bulk resistance from 92 Ω to 14.8 MΩ of the film from the obtained rGO.

A Bio-inspired Multifunctionalized Silk Fibroin.

A bio-inspired multifunctionalized silk fibroin (BMS) was synthesized in order to mimic the interaction of nidogen with the type IV collagen and laminin of basement membranes. The designed BMS consists of a motif of laminin α-chain-derived, called IK peptide, and type IV collagen covalently bound to the silk fibroin (SF) by using EDC/NHS coupling and a Cu-free click chemistry reaction, respectively. Silk fibroin was chosen as the main component of the BMS because it is versatile and biocompatible, induces an in vivo favorable bioresponse, and moreover can be functionalized with different methods. The chemical structure of BMS was analyzed by using X-ray photoelectron spectroscopy, attenuated total reflection-Fourier transform infrared, cross-polarization magic angle spinning nuclear magnetic resonance techniques, and colorimetric assay. The SF and BMS solutions were cross-linked by sonication to form hydrogels or casted to make films in order to evaluate and compare the early adhesion and viability of MRC5 cells. BMS hydrogels were also characterized by rheological and thermal analyses.

Free-Standing Graphene Oxide and Carbon Nanotube Hybrid Papers with Enhanced Electrical and Mechanical Performance and Their Synergy in Polymer Laminates.

Hybrid nanomaterials fabricated by the heterogeneous integration of 1D (carbon nanotubes) and 2D (graphene oxide) nanomaterials showed synergy in electrical and mechanical properties. Here, we reported the infiltration of carboxylic functionalized single-walled carbon nanotubes (C-SWNT) into free-standing graphene oxide (GO) paper for better electrical and mechanical properties than native GO. The stacking arrangement of GO sheets and its alteration in the presence of C-SWNT were comprehensively explored through scanning electron microscopy, X-ray photoelectron spectroscopy (XPS) and X-ray diffraction. The C-SWNTs bridges between different GO sheets produce a pathway for the flow of electrical charges and provide a tougher hybrid system. The nanoscopic surface potential map reveals a higher work function of the individual functionalised SWNTs than surrounded GO sheets showing efficient charge exchange. We observed the enhanced conductivity up to 50 times and capacitance up to 3.5 times of the hybrid structure than the GO-paper. The laminate of polystyrene composites provided higher elastic modulus and mechanical strength when hybrid paper is used, thus paving the way for the exploitation of hybrid filler formulation in designing polymer composites.

Ultrasound-Assisted Hydroxyapatite-Decorated Breath-Figure Polymer-Derived Ceramic Coatings for Ti6Al4V Substrates.

The introduction of nanoparticles (NPs) into the breath-figure-templated self-assembly (BFTSA) process is an increasingly common method to selectively decorate a surface porous structure. In the field of prosthetic devices, besides controlling the morphology and roughness of the structure, NPs can enhance the osteointegration mechanism because of their specific ion release. Among the most widely used NPs, there are silica and hydroxyapatite (HAp). In this work, we propose a novel one-stage method to fabricate NP-decorated surface porous structures that are suitable for prosthetic coating applications. This technique combines the classical direct BFTSA process with the cavitation effect induced by an ultrasonic atomizer that generates a mist of water droplets with embedded NPs. Coatings were successfully obtained by combining a UV cross-linkable polymer precursor, alkoxy silicone, with synthesized HAp NPs, on Ti6Al4V alloy discs. The cross-linked polymeric surface porous structures at selected concentrations were then pyrolyzed in an ammonia atmosphere to obtain a silicon oxynitride (SiON) ceramic coating. Herein, we report the chemical and morphological analyses of both the polymeric and ceramic coatings as well as the effect of NPs at the interface.

Amination of Graphene Oxide Leads to Increased Cytotoxicity in Hepatocellular Carcinoma Cells.

Clinically, there is an urgent need to identify new therapeutic strategies for selectively treating cancer cells. One of the directions in this research is the development of biocompatible therapeutics that selectively target cancer cells. Here, we show that novel aminated graphene oxide (haGO-NH2) nanoparticles demonstrate increased toxicity towards human hepatocellular cancer cells compared to pristine graphene oxide(GO). The applied novel strategy for amination leads to a decrease in the size of haGO-NH2 and their zeta potential, thus, assuring easier penetration through the cell membrane. After characterization of the biological activities of pristine and aminated GO, we have demonstrated strong cytotoxicity of haGO-NH2 toward hepatic cancer cells - HepG2 cell line, in a dose-dependent manner. We have presented evidence that the cytotoxic effects of haGO-NH2 on hepatic cancer cells were due to cell membrane damage, mitochondrial dysfunction and increased reactive oxygen species (ROS) production. Intrinsically, our current study provides new rationale for exploiting aminated graphene oxide as an anticancer therapeutic.

Strengthening of Wood-Like Materials via Densification and Nanoparticle Intercalation.

Recently, several chemical and physical treatments were developed to improve different properties of wood. Such treatments are applicable to many types of cellulose-based materials. Densification leads the group in terms of mechanical results and comprises a chemical treatment followed by a thermo-compression stage. First, chemicals selectively etch the matrix of lignin and hemicellulose. Then, thermo-compression increases the packing density of cellulose microfibrils boosting mechanical performance. In this paper, in comparison with the state-of-the-art for wood treatments we introduce an additional nano-reinforcemeent on densified giant reed to further improve the mechanical performance. The modified nanocomposite materials are stiffer, stronger, tougher and show higher fire resistance. After the addition of nanoparticles, no relevant structural modification is induced as they are located in the gaps between cellulose microfibrils. Their peculiar positioning could increase the interfacial adhesion energy and improve the stress transfer between cellulose microfibrils. The presented process stands as a viable solution to introduce nanoparticles as new functionalities into cellulose-based natural materials.

Aminated Graphene Oxide as a Potential New Therapy for Colorectal Cancer.

Nanotechnology-based drug delivery systems for cancer therapy are the topic of interest for many researchers and scientists. Graphene oxide (GO) and its derivates are among the most extensively studied delivery systems of this type. The increased surface area, elevated loading capacity, and aptitude for surface functionalization together with the ability to induce reactive oxygen species make GO a promising tool for the development of novel anticancer therapies. Moreover, GO nanoparticles not only function as effective drug carriers but also have the potential to exert their own inhibitory effects on tumour cells. Recent results show that the functionalization of GO with different functional groups, namely, with amine groups, leads to increased reactivity of the nanoparticles. The last steers different hypotheses for the mechanisms through which this functionalization of GO could potentially lead to improved anticancer capacity. In this research, we have evaluated the potential of amine-functionalized graphene oxide nanoparticles (GO-NH2) as new molecules for colorectal cancer therapy. For the purpose, we have assessed the impact of aminated graphene oxide (GO) sheets on the viability of colon cancer cells, their potential to generate ROS, and their potential to influence cellular proliferation and survival. In order to elucidate their mechanism of action on the cellular systems, we have probed their genotoxic and cytostatic properties and compared them to pristine GO. Our results revealed that both GO samples (pristine and aminated) were composed of few-layer sheets with different particle sizes, zeta potential, and surface characteristics. Furthermore, we have detected increased cyto- and genotoxicity of the aminated GO nanoparticles following 24-hour exposure on Colon 26 cells. The last leads us to conclude that exposure of cancer cells to GO, namely, aminated GO, can significantly contribute to cancer cell killing by enhancing the cytotoxicity effect exerted through the induction of ROS, subsequent DNA damage, and apoptosis.

Synthesis of Gold Nanoparticles Decorated with Multiwalled Carbon Nanotubes (Au-MWCNTs) via Cysteaminium Chloride Functionalization.

Gold nanoparticles (AuNPs) decorated CNTs are promising materials for photocatalytics and biosensors. However, the synthesis of AuNPs chemically linked to the walls of MWCNTs is challenging and toxic products such as thionylchloride (SOCl2) or [1-ethyl-3(dimethyl-amino) propyl] carbodiimide hydrochloride (EDAC) need to be used. This work reports a new approach to prepare gold nanoparticles decorated multiwalled carbon nanotubes (MWCNTs) by using cysteaminium chloride via the formation of a Zwitterionic acide-base bond. The grafting process consists of 3 mains steps: oxidation, thiolation and decoration of AuNPs on the surface of MWCNTs. The completion of each step has been verified out by both spectroscopic (Raman, UV-Vis, FT-IR) and Scanning Electron Miscroscopy (SEM). The chemical bonding states of synthesized products have been proven by X-ray photoelectron spectroscopy (XPS).

A mechanical system for tensile testing of supported films at the nanoscale.

Standard tensile tests of materials are usually performed on freestanding specimens. However, such requirement is difficult to implement when the materials of interest are of nanoscopic dimensions due to problems related to their handling and manipulation. In the present paper, a new device is presented for tensile testing of thin nanomaterials, which allows tests to be carried out on specimens initially deposited onto a macroscopic pre-notched substrate. On loading, however, no substrate effects are introduced, allowing the films to be freely stretched. The results obtained from a variety of thin metal or polymeric films are very promising for the further development of this technique as a standard method for nanomaterial mechanical testing.

Surface modification and characterization of GO/polymer thin coatings as excellent bio-active platforms for tissue regeneration.

Osteo-integration and tissue regeneration are vital for the longevity, durability, and unremitting functionality of medical implants/scaffolds implanted in vivo. It's essential for biomaterials used for in vivo implantation to induce the cellular secretion of growth factors, necessary for the desired tissue generation, since the administration of artificial growth factors, in vivo, is largely prohibited. Plasma functionalized (N2 and O2) and stabilized Graphene Oxide (GO) thin layers in a hybrid with amorphous carbon (aC) induced the expression of vascular endothelial growth factor (VEGF) and osteoprotegerin (OPG) growth factors in fibroblasts (hGF) and, more remarkably, in osteoblasts (hFOB) cells confirming the suitability for tissue regeneration and osteo-integration applications. We also observed a negative trend between hGF fibroblasts, but not hFOB osteoblasts, cellular viability and GO presence in the hybrid films that might indicate the phenomenon of oxidative stress. We traced that back to the presence of higher concentrations of carboxyl and the carbonyl groups on the surface of the GO rich coatings. The above described properties provided by GO coatings might be desirable for bio-selectivity applications and for the reduction of the undesired fibrosis process that is associated with medical implants in vivo environment. Moreover, novel plasma functionalized GO/polymer hybrid thin coating hybrid compositions are promising candidates for tissue engineering and bioengineering applications as excellent antimicrobial and anticancer platforms.