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Graphene Oxide Touches Blood: In Vivo Interactions of Bio-Coronated 2D Materials
Abstract
Graphene oxide is the hot topic of biomedical and pharmaceutical research of this decade. However, its complex interactions with human blood components complicate a transition of the promising in vitro results to clinical settings. Even if graphene oxide is made with same atoms our organs, tissues, and cells, its bi-dimensional nature causes unique interactions with blood proteins and biological membranes and can lead to severe effects like thrombogenicity and immune cells activation. In this review, we will describe the journey of graphene oxide after injection in the bloodstream, from the initial interactions with plasma proteins to the formation of the “biomolecular corona”, and biodistribution. We will consider the link between the chemical properties of graphene oxide (and its functionalized/reduced derivatives), protein binding and in vivo response. We will also summarize data on biodistribution and toxicity in view of current knowledge of biomolecular corona influence on these processes. Our aim is to shed light on unsolved problems of graphene oxide corona literature to build the groundwork for future drug delivery technology development.
... However, there is insufficient knowledge regarding the nature of the nanobiointeractions of GO with blood biological entities as well as possible nanotoxicity and adverse effects [24][25][26][27][28]. One method commonly used to assess the toxicity of GO to red blood cells (RBCs) is the hemolytic assay [29][30][31]. Furthermore, GO interacts with plasma proteins, leading to a protein coating commonly designated the protein corona [30,32]. ...
... One method commonly used to assess the toxicity of GO to red blood cells (RBCs) is the hemolytic assay [29][30][31]. Furthermore, GO interacts with plasma proteins, leading to a protein coating commonly designated the protein corona [30,32]. Indeed, the protein corona on the GO surface is a new biological identity that drives further nanobiointeractions, impacting its toxicity and biocompatibility [30,32]. ...
... Furthermore, GO interacts with plasma proteins, leading to a protein coating commonly designated the protein corona [30,32]. Indeed, the protein corona on the GO surface is a new biological identity that drives further nanobiointeractions, impacting its toxicity and biocompatibility [30,32]. ...
In this work, we performed an integrated study on the physicochemical changes of graphene oxide (GO) during the drying process in terms of their biological effects on red blood cells (hemolysis) and interactions with human plasma (protein corona formation). GO in aqueous dispersion (GO-Disp) was dried exploring two procedures: using a vacuum system at room temperature (GO-VD) and lyophilization (GO-LP). The nanomaterials were well characterized by microscopic (TEM, SEM, and AFM), spectroscopic (FTIR, UV–Vis, Raman, and ¹³C NMR), and XRD techniques. The lyophilization process produced a nanomaterial with a three-dimensional porous macrostructure and the lowest oxidation degree. In contrast, the vacuum-drying process at room temperature provided a nanomaterial with a film-like macrostructure, presenting a higher oxidation degree as well as physicochemical properties more similar to those of GO-Disp. All of the nanomaterials adsorbed human plasma proteins; however, the protein adsorption was more selective for GO-Disp. GO-VD induced hemolysis of red blood cells in a lower concentration than GO-Disp and GO-LP, but the protein corona formation suppressed the hemolytic effect for all nanomaterials. Finally, our results indicate that the method applied to dry GO nanomaterials has a critical influence on their nanobiointeractions with cells and proteins, suggesting that more attention should be paid to biomedical applications and toxicological evaluations associated with these promising nanomaterials.
Graphical abstract
... The GO oxygen groups' amount and quality can be varied by several reduction methods, to modify surface availability and, consequently, biological effects. Indeed, the attachment of macromolecules such as proteins and carbohydrates to GO is influenced by charge and steric hindrance of functional groups [7]. Since prokaryotic and eukaryotic cells respond to the adsorbed macromolecules, oxygen groups will eventually determine GO effects on bacteria and human cells in biological media [7][8][9]. ...
... Indeed, the attachment of macromolecules such as proteins and carbohydrates to GO is influenced by charge and steric hindrance of functional groups [7]. Since prokaryotic and eukaryotic cells respond to the adsorbed macromolecules, oxygen groups will eventually determine GO effects on bacteria and human cells in biological media [7][8][9]. ...
Graphene Oxide (GO) is the oxidized form of graphene rich in surface groups comprising carbonyl, carboxyl, hydroxyl, and epoxy residues. The solubility in water makes GO an ideal material in the biomedical field though the plethora of synthesis methods available can modify the balance of oxygen groups and, consequently, the effects on eukaryotic and prokaryotic cells. For this reason, GO materials are always characterized with spectroscopic methods such as X-ray photoelectron and Fourier-transform Infrared spectroscopy. However, these techniques have some limitations, being disruptive and not clearly indicating oxygen functionalities available to react with polymers or biological media.
In this work, we exploit GO reactivity with copper ions to develop a colorimetric method for the facile evaluation of surface oxidation degree and accessibility in polymeric samples. In the presence of GO, Cu²⁺ is reduced to Cu¹⁺ and can react with bicinchoninic acid and induce light absorption at 562 nm. We observed that this reaction is dependent both on concentration and oxidation degree, and can be used to estimate GO exposure in thick composite samples. This technique will be fundamental in the future for scaffold characterization in tissue engineering and all the surface science studies analyzing GO-related materials' interactions with biological entities.
... The protein corona is one of the first factors to be addressed, which causes the rapid change of GO in the bloodstream [164]. This protein coating induces the size change of GO that substantially influences the interactions with the cells [34]. ...
... The body's non-specific immune defense relies on phagocytosis of foreign molecules via macrophages, providing a significant barrier to intravenous injections of GO nanocarriers. In addition, GO nanocarriers have a high probability of being removed by macrophages before reaching their destination and may initiate an inflammatory reaction [24,164]. In particular, when in the bloodstream, larger nanoparticles (>200 nm) have a higher risk of being recognized and sequestered [44,53,123,137] by the phagocytosing cells of the immune system (i.e., macrophages, dendritic cells, neutrophils, and B lymphocytes), which are responsible for recognizing the foreigner and destroying it via enzymatic digestion, as explained previously [31]. ...
Functionalized graphene oxide (GO) nanoparticles are being increasingly employed for designing modern drug delivery systems because of their high degree of functionalization, high surface area with exceptional loading capacity, and tunable dimensions. With intelligent controlled release and gene silencing capability, GO is an effective nanocarrier that permits the targeted delivery of small drug molecules, antibodies, nucleic acids, and peptides to the liquid or solid tumor sites. However, the toxicity and biocompatibility of GO-based formulations should be evaluated, as these nanomaterials may introduce aggregations or may accumulate in normal tissues while targeting tumors or malignant cells. These side effects may potentially be impacted by the dosage, exposure time, flake size, shape, functional groups, and surface charges. In this review, the strategies to deliver the nucleic acid via the functionalization of GO flakes are summarized to describe the specific targeting of liquid and solid breast tumors. In addition, we describe the current approaches aimed at optimizing the controlled release towards a reduction in GO accumulation in non-specific tissues in terms of the cytotoxicity while maximizing the drug efficacy. Finally, the challenges and future research perspectives are briefly discussed.
... Over the past decade, graphene oxide has been the most investigated for its amazing properties. Many studies prove that graphene-oxide has a high surface area [29][30][31][32], many functional groups [33][34][35], protein adsorption [36][37][38][39], antimicrobial potentials [40][41][42][43][44][45], hydrophilicity properties [34], [44,45] and flexible to handle, so they were used for wound dressing to prevent infection in different wounds [41], [46][47][48][49][50][51][52]. When addition small quantity of GO (< 5%) enhanced the mechanical properties of the wound dressing and protein affinity which wound fast healing, and support of bone tissue generation. ...
... The nomenclature given to GBM as well as the various methods of synthesis denote a source of misperception since, in some scientific works, what authors describe as graphene/functionalized graphene is another member of this ultrathin carbon family and this leads to (sometimes apparent) contradictions in the results ascribed to the same GBM [22,23]. If we try to narrow down the specific GO family, the effects of GO on biological systems are often compared to its reduced form, named reduced graphene oxide (rGO). ...
Graphene oxide (GO) nanoparticles, due to their favorable water solubility, compared to graphene (GA), are a hot research topic in biomedical and pharmaceutical research. However, GO clinical translation may be complicated by its high surface/volume ratio enhancing the interaction with human blood components. In fact, GO’s bi-dimensional nature and strong negative charge may lead to severe biological effects, such as thrombogenicity and immune cell activation. This study explores the impact of further GO surface chemical modulation on major adverse effects: blood plasma coagulation and hemolysis. To this aim, we refined GO nanoparticles by fine-tuned reduction chemistry, esterification and introduction of negative or positive charges. With this approach, we were able to mitigate plasma coagulation and hemolysis at variable degrees and to identify GO derivatives with improved biocompatibility. This opens the door to the progress of graphene-based nanotheranostic applications.
... 9 Graphene oxide (GO), a derivative of graphene, has gathered substantial attention due to its remarkable features and versatile applications in the biomedical field. [10][11][12][13] GO is a well-known photosensitizing agent, that can absorb near-infrared (NIR) light and convert it efficiently into heat. [14][15][16] Moreover, GO demonstrates excellent capability to generate reactive oxygen species (ROS) upon light irradiation, promoting selective destruction of tumor cells while minimizing damage to healthy tissues. ...
Surgically addressing tumors poses a challenge, requiring a tailored, multidisciplinary approach for each patient based on the unique aspects of their case. Innovative therapeutic regimens combined to reliable reconstructive methods can contribute to an extended patient's life expectancy. This study presents a detailed comparative investigation of near-infrared therapy protocols, examining the impact of non-fractionated and fractionated irradiation regimens on cancer treatment. The therapy is based on the implantation of graphene oxide/poly(lactic-co-glycolic acid) three-dimensional printed scaffolds, exploring their versatile applications in oncology by the examination of pro-inflammatory cytokine secretion, immune response, and in vitro and in vivo tumor therapy. The investigation into cell death patterns (apoptosis vs necrosis) underlines the pivotal role of protocol selection underscores the critical influence of treatment duration on cell fate, establishing a crucial parameter in therapeutic decision-making. In vivo experiments corroborated the profound impact of protocol selection on tumor response. The fractionated regimen emerged as the standout performer, achieving a substantial reduction in tumor size over time, surpassing the efficacy of the non-fractionated approach. Additionally, the fractionated regimen exhibited efficacy also in targeting tumors in proximity but not in direct contact to the scaffolds. Our results address a critical gap in current research, highlighting the absence of a standardized protocol for optimizing the outcome of photodynamic therapy. The findings underscore the importance of personalized treatment strategies in achieving optimal therapeutic efficacy for precision cancer therapy.
... On the basis of the excellent targeting efficacy offered by the surface chemistry of CNMs at the nanobiointerface, it is reasonable to anticipate that CNM-based drug delivery platforms will promote the targeted theranostics through their practical applications. However, upon contact with biofluids, CNMs tend to adsorb a variety of proteins [71] including serum albumin, immunoglobulin, coagulation proteins, lipoproteins, etc., to form a coating layer known as protein corona (Fig. 5a). It is well known that the proteins in this layer alter the surface chemistry of CNMs and form a new interface between ...
Carbon nanomaterials (CNMs), including fullerenes, carbon nanotubes, graphene, nanodiamonds and their derivatives, have been extensively used for biomedical applications, especially for cancer diagnosis and treatment. Most of these applications require covalent reactions to give the CNMs a high dispersibility in physiological environments, new biological functions for diagnosis and treatment, and tailored interactions with biological systems for efficient therapy. In this review, we summarize: the covalent bonding approaches to functionalize CNMs with polar surface groups, hydrophilic polymer layers, and bioactive molecules, and the specific surface chemistry that can prevent the interactions of CNMs with plasma proteins to suppress non-specific uptake and to enhance targeted uptake, aiming to advance the design of CNM-based therapeutics for precise theranostics.
... Moreover, various studies have reported that the GO contributes to the increase in cell viability [76,77]. Similarly, PLLA polymers have shown promising results when applied to bone scaffolds, as they increase the proliferation of various cell types [78,79]. ...
Accurately printing customizable scaffolds is a challenging task because of the complexity of bone tissue composition, organization, and mechanical behavior. Graphene oxide (GO) and poly-L-lactic acid (PLLA) have drawn attention in the field of bone regeneration. However, as far as we know, the Fischer–Koch model of the GO/PLLA association for three-dimensional (3D) printing was not previously reported. This study characterizes the properties of GO/PLLA-printed scaffolds in order to achieve reproducibility of the trabecula, from virtual planning to the printed piece, as well as its response to a cell viability assay. Fourier-transform infrared and Raman spectroscopy were performed to evaluate the physicochemical properties of the nanocomposites. Cellular adhesion, proliferation, and growth on the nanocomposites were evaluated using scanning electron microscopy. Cell viability tests revealed no significant differences among different trabeculae and cell types, indicating that these nanocomposites were not cytotoxic. The Fischer Koch modeling yielded satisfactory results and can thus be used in studies directed at diverse medical applications, including bone tissue engineering and implants.
... However, there are discrepancies in the published data on biodistribution and excretion pathways at the level of animal research. These variances are seen in a variety of organs and are directly related to the specific features of the synthesized NGOs [30, 46,51,[54][55][56]. This heterogeneity in findings underscores the complex interplay between the properties of the nanomaterials and their interaction with biological systems. ...
Graphene, fullerenes, diamond, carbon nanotubes, and carbon dots are just a few of the carbon-based nanomaterials that have gained enormous popularity in a variety of scientific disciplines and industrial uses. As a two-dimensional material in the creation of therapeutic delivery systems for many illnesses, nanosized graphene oxide (NGO) is now garnering a large amount of attention among these materials. In addition to other benefits, NGO functions as a drug nanocarrier with remarkable biocompatibility, high pharmaceutical loading capacity, controlled drug release capability, biological imaging efficiency, multifunctional nanoplatform properties, and the power to increase the therapeutic efficacy of loaded agents. Thus, NGO is a perfect nanoplatform for the development of drug delivery systems (DDSs) to both detect and treat a variety of ailments. This review article’s main focus is on investigating surface functionality, drug-loading methods, and drug release patterns designed particularly for smart delivery systems. The paper also examines the relevance of using NGOs to build DDSs and considers prospective uses in the treatment of diseases including cancer, infection by bacteria, and bone regeneration medicine. These factors cover the use of naturally occurring medicinal substances produced from plant-based sources.
... Notably, a study by Maryam Maghsudi et al. explored the interaction between silver nanoclusters (Ag NCs) and human blood plasma proteins and revealed alterations in the physical and chemical properties of both proteins and Ag NCs (Maghsudi et al., 2018). Similarly, nanomaterials characterized by exceptionally large surface areas, such as graphene and its derivatives, were also susceptible to protein adsorption, resulting in compromised functionalization (Palmieri et al., 2019). ...
Microbial infections continue to pose a significant health challenge, especially with an increase in drug-resistant bacteria. Conventional antibiotic treatments show limited efficacy, prompting researchers to explore alternative treatments. Photodynamic therapy (PDT) has emerged as a promising alternative that uses reactive oxygen species (ROS) to induce oxidative stress, offering the potential for cyclic treatment without fostering new drug resistance mechanisms. The success of PDT relies heavily on the selection of appropriate photosensitizers (PSs). Various nanomaterials are being developed as PSs or carriers to enhance the efficacy of PDT in the antibacterial field. In this comprehensive review, we discuss the four main ROS generated during PDT and outline their corresponding antibacterial mechanisms. Additionally, we highlight the prominent types of nanomaterials used as PSs or carriers in PDT. We analyze the current challenges associated with nanomaterial-based PDT for antibacterial therapy and propose potential strategies for optimizing their applications.
... Exploring the bionano interactions with the biological milieu has therefore emerged as the missing link between benchtop discoveries and the clinical applicability of nanomedicines. The formation of PC on graphene-based materials has been the subject of recent studies [20,21]. For instance, Liu et al. studied the influence of HSA on GO surface at different pH values and demonstrated that the attachment of GO to a model cell membrane was reduced in the presence of HSA corona [22]. ...
Graphene-based nanomaterials have attracted significant attention in the field of nanomedicine due to their unique atomic arrangement which allows for manifold applications. However, their inherent high hydrophobicity poses challenges in biological systems, thereby limiting their usage in biomedical areas. To address this limitation, one approach involves introducing oxygen functional groups on graphene surfaces, resulting in the formation of graphene oxide (GO). This modification enables improved dispersion, enhanced stability, reduced toxicity, and tunable surface properties. In this review, we aim to explore the interactions between GO and the biological fluids in the context of theranostics, shedding light on the formation of the “protein corona” (PC) i.e., the protein-enriched layer that formed around nanosystems when exposed to blood. The presence of the PC alters the surface properties and biological identity of GO, thus influencing its behavior and performance in various applications. By investigating this phenomenon, we gain insights into the bio-nano interactions that occur and their biological implications for different intents such as nucleic acid and drug delivery, active cell targeting, and modulation of cell signalling pathways. Additionally, we discuss diagnostic applications utilizing biocoronated GO and personalized PC analysis, with a particular focus on the detection of cancer biomarkers. By exploring these cutting-edge advancements, this comprehensive review provides valuable insights into the rapidly evolving field of GO-based nanomedicine for theranostic applications.
Graphical Abstract
... As an important derivative of graphene, a new 2D nanomaterial, GO has aroused widespread interest in the field of biomedicine, and has become one of the research hotspots in nanobiomedicine, especially nano-drug delivering [174]. The main advantages of GO [175] as a nano-drug loading system include: (1) super-large specific surface area, which can achieve ultra-high drug loading rate; (2) strong targeting and easy enrichment in tumor sites; (3) functionalized GO has good biocompatibility and stability under physiological conditions. ...
Two-dimensional (2D) nanomaterials, such as graphene, black phosphorus and transition metal dichalcogenides, have attracted increasing attention in biology and biomedicine. Their high mechanical stiffness, excellent electrical conductivity, optical transparency, and biocompatibility have led to rapid advances. Neuroscience is a complex field with many challenges, such as nervous system is difficult to repair and regenerate, as well as the early diagnosis and treatment of neurological diseases are also challenged. This review mainly focuses on the application of 2D nanomaterials in neuroscience. Firstly, we introduced various types of 2D nanomaterials. Secondly, due to the repairment and regeneration of nerve is an important problem in the field of neuroscience, we summarized the studies of 2D nanomaterials applied in neural repairment and regeneration based on their unique physicochemical properties and excellent biocompatibility. We also discussed the potential of 2D nanomaterial-based synaptic devices to mimic connections among neurons in the human brain due to their low-power switching capabilities and high mobility of charge carriers. In addition, we also reviewed the potential clinical application of various 2D nanomaterials in diagnosing and treating neurodegenerative diseases, neurological system disorders, as well as glioma. Finally, we discussed the challenge and future directions of 2D nanomaterials in neuroscience.
Graphical Abstract
... Recent advances on the exploration of GDMs, including graphene oxide (GO), on immune cell interactions induce important immunosuppressive actions over different myeloid cells 18 , being more extensively explored in the case of macrophages 19 . Particularly, GO seems to have an outstanding ability to induce macrophage cytotoxicity, but also to alter their phagocytic capacity and, very importantly, polarize them towards either pro-or anti-inflammatory phenotypes by manipulating different physico-chemical properties of this nanomaterial such as its oxidation degree 20,21 . ...
Multiple Sclerosis (MS) is a chronic, inflammatory disease of the central nervous system. Despite the pharmacological arsenal approved for MS, there are treatment-reluctant patients for whom cell therapy appears as the only therapeutic alternative. Myeloid-derived suppressor cells (MDSCs) are immature cells of the innate immune response able to immunosuppress T lymphocytes and to promote oligodendroglial differentiation in experimental autoimmune encephalomyelitis (EAE), a preclinical model for MS. Culture devices need to be designed so that MDSCs maintain a state of immaturity and immunosuppressive function similar to that exerted in the donor organism. Graphene oxide (GO) has been described as a biocompatible material with the capacity to biologically modulate different cell types, including immune cells. In the present work, we show how MDSCs isolated from immune organs of EAE mice maintain an immature phenotype and highly immunosuppressive activity on T lymphocytes after being cultured on 2D reduced GO films (rGO200) compared to those grown on glass. This activity is depleted when MDSCs are exposed to slightly rougher and more oxidized GO substrates (rGO90). The greater reduction in cell size of cells exposed to rGO90 compared to rGO200 is associated with the activation of apoptosis processes. Taken together, the exposure of MDSCs to GO substrates with different redox state and roughness appears as a good strategy to control MDSC activity in vitro. This versatility of GO nanomaterials and the impact of their physico-chemical properties in immunomodulation open the door to its possible selective therapeutic use for pathologies where MDSCs need to be enhanced or inhibited.
... Surface-charged fullerenes can inhibit the growth of M. avium and Mtb [11], GO in a reduced state exerts antibacterial effect against Mtb [13] and GO-Ethambutol particles inhibited M. smegmatis growth in axenic liquid culture [14,15]. It shall be noted that in many of these experiments functionalized forms rather than a "pure form" of GO were used, with the functional groups providing peculiar physical and chemical features [16]. Interestingly, the investigators focused their studies mainly at evaluating the activity of these CNM on bacteria, paying less attention to the effect of these molecules on eukaryotic cells. ...
Graphene Oxide has been proposed as a potential adjuvant to develop improved anti-TB treatment, thanks to its activity in entrapping mycobacteria in the extracellular compartment limiting their entry in macrophages. Indeed, when administered together with linezolid, Graphene Oxide significantly enhanced bacterial killing due to the increased production of Reactive Oxygen Species. In this work, we evaluated Graphene Oxide toxicity and its anti-mycobacterial activity on human peripheral blood mononuclear cells. Our data show that Graphene Oxide, different to what is observed in macrophages, does not support the clearance of Mycobacterium tuberculosis in human immune primary cells, probably due to the toxic effects of the nano-material on monocytes and CD4+ lymphocytes, which we measured by cytometry. These findings highlight the need to test GO and other carbon-based nanomaterials in relevant in vitro models to assess the cytotoxic activity while measuring antimicrobial potential.
... Futhermore, oxygen-containing functional groups on the GO surface and polymer matrix can increase the interfacial contact force via hydrogen bonding [17]. In this study, an interface for the conversion of 3D nanoprobes to 2D was designed utilizing GO's distinct nanostructure, large surface area, solubility in aqueous solution, and ease of modification of oxygen-containing functional groups [18]. Few comparative interface contact area studies have been undertaken in prior reports (mainly for biosensing applications [19]). ...
The electrical characteristic of cancer cells is neglected among tumor biomarkers. The development of nanoprobes with opposing charges for monitoring the unique electrophysiological characteristics of cancer cells. Micro-nano size adsorption binding necessitates consideration of the nanoprobe’s specific surface area. On the basis of the electrophysiological characteristics of circulating tumor cells (CTCs), clinical application and performance assessment are determined. To demonstrate that cancer cells have a unique pattern of electrophysiological patterns compared to normal cells, fluorescent nanoprobes with opposing charges were developed and fabricated. Graphene oxide (GO) was used to transform three-dimensional (3D) nanoprobes into two-dimensional (2D) nanoprobes. Compare 2D and 3D electrophysiological magnetic nanoprobes (MNP) in clinical samples and evaluate the adaptability and development of CTCs detection based on cell electrophysiology. Positively charged nanoprobes rapidly bind to negatively charged cancer cells based on electrostatic interactions. Compared to MNPs(+) without GO, the GO/MNPs(+) nanoprobe is more efficient and uses less material to trap cancer cells. CTCs can be distinguished from normal cells that are fully unaffected by nanoprobes by microscopic cytomorphological inspection, enabling the tracking of the number and pathological abnormalities of CTCs in the same patient at various chemotherapy phases to determine the efficacy of treatment. The platform for recognizing CTCs on the basis of electrophysiological characteristics compensates for the absence of epithelial biomarker capture and size difference capture in clinical performance. Under the influence of electrostatic attraction, the binding surface area continues to influence the targeting of cancer cells by nanoprobes. The specific recognition and detection of nanoprobes based on cell electrophysiological patterns has enormous potential in the clinical diagnosis and therapeutic monitoring of cancer.
... 269 Notably, studies have shown that graphene quantum dots (GQDs) are less toxic than graphene oxide (GO). 270,271 Sun et al constructed an antibacterial system by combining GQDs and low-concentration H 2 O 2 . 272 During the reaction, the POD-like activity of GQDs converts H 2 O 2 into -OH, which improves the antibacterial properties and avoids unnecessary damage caused by the use of high concentrations of H 2 O 2 . ...
Bacterial-infected wounds are a serious threat to public health. Bacterial invasion can easily delay the wound healing process and even cause more serious damage. Therefore, effective new methods or drugs are needed to treat wounds. Nanozyme is an artificial enzyme that mimics the activity of a natural enzyme, and a substitute for natural enzymes by mimicking the coordination environment of the catalytic site. Due to the numerous excellent properties of nanozymes, the generation of drug-resistant bacteria can be avoided while treating bacterial infection wounds by catalyzing the sterilization mechanism of generating reactive oxygen species (ROS). Notably, there are still some defects in the nanozyme antibacterial agents, and the design direction is to realize the multifunctionalization and intelligence of a single system. In this review, we first discuss the pathophysiology of bacteria infected wound healing, the formation of bacterial infection wounds, and the strategies for treating bacterially infected wounds. In addition, the antibacterial advantages and mechanism of nanozymes for bacteria-infected wounds are also described. Importantly, a series of nanomaterials based on nanozyme synthesis for the treatment of infected wounds are emphasized. Finally, the challenges and prospects of nanozymes for treating bacterial infection wounds are proposed for future research in this field.
... framework the results become clearer. Due to the sharp edges of GO and RgO, hemolytic-effects might be expected in vivo, possibly caused by nano-material blades disrupting cell-membranes, as reported for the GO interactions with the bacteria" (Figure 9) [4]. ...
This work starts after seeing a recent open letter for transparency related production a quality control technique of m RNA vaccine first signed by Tarro G, Luisetto M and Monsellato ML and an Editorial recognized by IMA Marijnskaya academy: Graphene and Derivates: Physico-Chemical and Toxicology properties in the m-RNA Vaccine Manufacturing Strategy Needed specific proof of absence for the regulatory aspects (accepted for publication). Other relevant evidences come from the work of Giovannini et al. related Dark Field microscopy assay of the blood of 1086 symptomatic subjects after vaccination with two types of mRNA vaccine of great interest on this field the work of Campra P and Young RO, Young Me Lee or Ki-Yeob J. The Aim of this work is to investigate the self-auto assembly properties of graphene and derivates in order to Find relationship in some biotechnological application like mRNA vaccine. After a review part an experimental hypotesys project will be submitted to the researcher to produce a global conclusion. The recent evidences published in last period induced the idea to more deeply study these properties for The clinico-toxicological aspects involved.
... Dendritic-cells fail to present antigens to lymphocytes when they uptake GO (2e). Lymphocyte activity is not inhibited, and BC protects lymphocytes from apoptosis (2f) [14]. ...
This work start after seeing an recent open letter for transparency related production an quality control technique of mRNA vaccine first signed by Tarro G, Luisetto M and Monsellato ML and an editorial recognized by IMA Marijnskaya academy: Graphene and Derivate: Physio-Chemical and Toxicology properties in the mRNA Vaccine Manufacturing Strategy, needed specific proof of absence for the regulatory aspects (accepted for publication). Other relevant evidences related to this topic comes from the work of Giovannini, et al. related Dark field microscope assay of the blood of 1086 symptomatic subjects after vaccination with two types of m-RNA vaccine of great interest on this field also the work of P Campra and Young RO, Young Me Lee or Ki-Yeob J. Observing all this recent evidences the aim of this work is to investigate not only the graphene presence (or not) in vials of mRNA Vaccine but also the self-auto assembly properties of graphene and derivate. This is in order to find relationship in some biotechnological application like m-RNA Vaccine's large scale production. After a review part an experimental hypothesis project will be submitted to the researcher to produce a global conclusion related the topic investigated. The recent evidences published induced the idea to more deeply study these properties for the clinico-toxicological aspects involved. The pro-coagulant properties of the coronavirus covid-19 Spike Protein are well knower by scientific literature as well as the toxicological profile of the graphene derivate. What can happen if this two substantive can act in patients with platelet disorder in the same time?
... It has been reported that GO attracts many of the adhesion molecules necessary for cell www.nature.com/scientificreports/ attachment and growth in culture medium, thus providing a biological environment to selectively increase cell activity [34][35][36] . ...
Antimicrobial surfactants contained in mouthrinse have excellent efficacy, but are not retained on the tooth surface (are rinsed away) due to their low water resistance and thus do not exhibit sustained antibacterial activity. We have developed a new coating method using graphene oxide (GO) that retains the surfactant on the tooth surface even after rinsing with water, thus providing a sustained antibacterial effect. Ultra-thin films of GO and an antimicrobial agent were prepared by (1) applying GO to the substrate surface, drying, and thoroughly rinsing with water to remove excess GO to form an ultrathin film (almost a monolayer, transparent) on the substrate surface, then (2) applying antimicrobial cationic surface active agents (CSAAs) on the GO film to form a composite coating film (GO/CSAA). GO/CSAA formation was verified by scanning electron microscopy, Raman spectroscopy, X-ray photoelectron spectroscopy, and ζ-potential and contact angle measurements. GO/CSAA was effective at inhibiting the growth of oral pathogens for up to 7 days of storage in water, and antibacterial activity was recovered by reapplication of the CSAA. Antibacterial GO/CSAA films were also formed on a tooth substrate. The results suggest that GO/CSAA coatings are effective in preventing oral infections.
... Bidimensional materials show different interesting physical properties, making them suitable for many potential applications, including energy storage [1][2][3], biomedical research [4][5][6], field-effect transistors (FETs) [7][8][9][10], as well as sensors and biosensing [11,12]. One of these materials is Molybdenum disulfide, MoS 2 , a layered dichalcogenide with a hexagonal structure reminiscent of graphene. ...
Using structural relaxation calculations and first-principles molecular dynamics (FPMD),
we performed numerical simulations to explore the interaction of a 2D MoS2 surface and a platinum atom, calculating the optical properties of the resulting material. We explored three initial positions for the interaction of the Pt atom and the pristine MoS2 surface, plus another position between Pt and the MoS2 surface with a sulfur vacancy VS. The surface absorbed the Pt atom in all cases considered, with absorption energies ranging from −2.77 eV to −5.83 eV. We calculated the optical properties and band structure of the two cases with the largest absorption energies (−3.45 eV and −5.83 eV). The pristine MoS2 is a semiconductor with a gap of around 1.80 eV. With the adsorption of the Pt atom (the −3.45 eV case), the material reduces its band gap to 0.95 eV. Additionally, the optical absorption in the visible range is greatly increased. The energy band structure of the 2D MoS2 with a sulfur vacancy VS shows a band gap of 0.74 eV, with consequent changes in its optical properties. After the adsorption of Pt atoms in the VS vacancy, the material has a band gap of 1.06 eV. In this case, the optical absorption in the visible range increases by about eight times. The reflectivity in the infrared range gets roughly doubled for both situations of the Pt-absorbed atom considered. Finally, we performed two FPMD
runs at 300 K to test the stability of the cases with the lowest and highest absorption energies observed, confirming the qualitative results obtained with the structural relaxations
... Graphene is a one-atom-thick two-dimensional material composed of honeycomb-type structure, sp2 hybridised carbon atoms. In a range of biological applications, graphene and graphene oxide (GO) have been utilised as antibacterial agents [94], diagnostic agent [95] and as scaffolds for bone regeneration [96]. In comparison to other organic dyes, GQDs have remarkable optical properties that are solely dependent on their shape, surface properties, and size which makes them ideal for bioimaging [97]. ...
Alzheimer's disease (AD) is one of the most common forms of dementia, affecting more than 50 million people globally. The onset of AD is linked to age, smoking, obesity, hypercholesterolemia, physical inactivity, depression, gender, and genetics of an individual. The accumulation of Aβ peptides and neurofibrillary tangles (NFTs) in the brain is one of the critical factors that lead to AD, which is known to disrupt neuronal signaling and causing neurodegeneration. As per the current understanding, inhibiting the accumulation of Aβ peptides and NFTs is crucial in the management/treatment of AD. Latest research studies show that nanoparticles have the potency of improving drug transport across the blood-brain barrier easily. Specifically, graphene quantum dots (GQDs), a type of semiconducting nanoparticles, have been established as effective inhibitors for blocking the aggregation of Aβ peptides. The small size of GQDs allows them to pass through the blood-brain barrier with ease. Moreover, GQDs have fluorescence properties, which can be used to detect the concentration of Aβ in vivo. In recent years, compared to other carbon materials, the low cytotoxicity and high biocompatibility of GQDs, give them an advantage in the suitability and clinical research for AD. In this manuscript, we have discussed the role of different types of nanoparticles in the transportation of encapsulated or co-assembled compound drugs for the treatment of AD and importantly, the role of GQDs in the diagnosis and management/treatment of AD.
... Bidimensional materials show different interesting physical properties, making them suitable for many potential applications, including energy storage [1][2][3], biomedical research [4][5][6], field-effect transistors (FETs) [7][8][9][10], as well as sensors and biosensing [11,12]. One of such materials is Molybdenum disulfide, MoS2, a layered dichalcogenide with a hexagonal structure similar to graphene. ...
Using first-principles molecular dynamics (FPMD), we performed numerical simulations at 300 K to explore the interaction of a 2D MoS2 surface and a platinum atom, calculating the optical properties of the resulting material. The pristine MoS2 is a semiconductor with a gap of around 1.8 eV. The Pt atom is chemisorbed by the surface with an adsorption energy of −1.718 eV. With the adsorption of the Pt atom, the material remains a semiconductor, and its energy band gap reduces to 1.04 eV. But changes in the material's energy band structure imply substantial changes in its optical properties. The energy band structure of the 2D MoS2 with a sulfur vacancy VS shows that the material becomes a conductor, and there are significant changes in its optical properties. We also found that the Pt atom chemisorbs in a sulfur vacancy of the material, with an adsorption energy of −4.1164 eV. After the adsorption of Pt atoms in the sulfur vacancy, the material becomes a semiconductor with a band gap of 1.06 eV, and the changes in the optical absorption and reflectivity are significant.
... De Paoli et al. (2014) reported that an IgG corona causes thrombocyte fragmentation, leading to induction of thrombocyte aggregation and release of platelet membrane microparticles. Palmieri et al. (2019) also extensively reviewed the adverse effects that protein corona coated graphene oxide has on blood integrity. Barbalinardo et al. (2018) reported that the cytotoxic activity of citrate-coated silver NPs toward NIH-3T3 cells was increased after protein corona formation, contrary to what was observed with CNTs (Gu et al. 2015) and graphene oxide (Hu et al. 2011). ...
The surfaces of nanoparticles become covered by biomolecules in biological fluids. This protein ‘corona’ modifies materials’ characteristics and biological activity. The composition of the protein corona is dynamic, abundant biomolecules that bind first are subsequently replaced by less abundant but more tightly bound ones. Here, we explore the formation of the silver nanoparticle protein corona on exposure to cell culture media containing 10% fetal bovine serum supplemented Dulbecco's Modified Eagle's medium. Sodium dodecyl-sulfate polyacrylamide gel electrophoresis and liquid chromatography-mass spectrometry/mass spectrometry analysis were used to monitor how different parameters such as incubation time, heating duration, cell culture medium, incubation temperature, and the number of washes affect the nanoparticle–protein corona complex. silver nanoparticles with and without bound proteins were characterized by electron microscopy, dynamic light scattering, and ultraviolet-visible-near-IR spectroscopy. The tetrazolium-based MTT assay was used to determine viability of A549 human lung adenocarcinoma cells treated with silver nanoparticles. Characterization of the nanoparticles before and after protein binding provided insights into their changing morphology on corona formation. Our results confirmed that the physiological environment directly affects protein corona formation on nanoparticle surfaces. In particular, incubation condition-dependent differences in the amount of bound proteins were observed. This work highlights the importance of environmental drivers of protein adsorption, which should be considered when predicting and/or controlling protein targets of silver nanoparticles.
... On their ''journey" through the body following i.v. injection, 2D materials will inevitably encounter and bind plasma proteins on their surface [130]. Growing evidence has been reported that pre-coating of nanomaterials with host proteins could enable regulatory control of nanomaterial-induced immune response, either preventing their rapid clearance by the mononuclear phagocyte system, for instance, by masking the complement-activating properties or boosting the immune response by coating with specific antibodies. ...
Two-dimensional (2D) materials such as the graphene-based materials, transition metal dichalcogenides, transition metal carbides and nitrides (MXenes), black phosphorus, hexagonal boron nitride, and others have attracted considerable attention due to their unique physicochemical properties. This is true not least in the field of medicine. Understanding the interactions between 2D materials and the immune system is therefore of paramount importance. Furthermore, emerging evidence suggests that 2D materials may interact with microorganisms – pathogens as well as commensal bacteria that dwell in and on our body. We discuss the interplay between 2D materials, the immune system, and the microbial world in order to bring a systems perspective to bear on the biological interactions of 2D materials. The use of 2D materials as vectors for drug delivery and as immune adjuvants in tumor vaccines, and 2D materials to counteract inflammation and promote tissue regeneration, are explored. The bio-corona formation on and biodegradation of 2D materials, and the reciprocal interactions between 2D materials and microorganisms, are also highlighted. Finally, we consider the future challenges pertaining to the biomedical applications of various classes of 2D materials.
... In spite of the large number of scientific studies on CNMs, often, it is difficult to anticipate which type is the best performer for specific applications, and even more so in the case of biological uses where there is a high degree of biomolecular complexity [27][28][29][30]. Interactions between CNMs and biomolecules, such as proteins [31] and DNA [32], are very relevant in this regard for their key roles in the determination of the dynamic corona on their surface [33], which, in turn, affects their fate in vivo [34], spanning their biodistribution [35] to immune response [36,37] and biodegradation [38,39]. ...
Carbon nanomaterials have attracted great interest for their unique physico-chemical properties for various applications, including medicine and, in particular, drug delivery, to solve the most challenging unmet clinical needs. Graphitization is a process that has become very popular for their production or modification. However, traditional conditions are energy-demanding; thus, recent efforts have been devoted to the development of greener routes that require lower temperatures or that use waste or byproducts as a carbon source in order to be more sustainable. In this concise review, we analyze the progress made in the last five years in this area, as well as in their development as drug delivery agents, focusing on active targeting, and conclude with a perspective on the future of the field.
In the antimicrobial resistance era, carbon‐based nanomaterials (CBNs) such as fullerenes, carbon dots, graphene, and their derivatives are promising therapeutic tools in combating viral diseases. This review shows that these materials have broad‐spectrum antiviral activity against 33 viruses belonging to different Baltimore groups. CBNs also exhibit antimicrobial activity against bacteria and fungi and possess a low risk of selecting for resistance, since their primary mode of antimicrobial action involves physically damaging the microbes. CBNs also offer additional promising properties, including enhanced antiviral effectiveness under diverse types of irradiation and facilitating antiviral immune responses. Their potential as antiviral agents is still in its infancy and future research should focus on their toxicity, antiviral mechanisms, pharmacokinetics, and bioavailability. They are also potential antiviral materials for preventing the transmission of viral diseases for use in face masks, shields, hospital and airport surfaces, and elevators, among others. It is anticipated that CBNs will play an increasingly significant role in the fight against viruses and infectious diseases.
The molecular layer that adsorbs on the biomaterial surface upon contacting body tissues and fluids, termed the conditioning layer (CL), influences cell behavior regulating scaffold integration and resilience in a patient-specific fashion. To predict and improve the clinical outcome of 3D-printed scaffolds, graphene coatings are employed in bone tissue engineering, due to the possibility to functionalize its chemical/physical properties. In this study, we investigated the composition and the influence of the CL on three different graphene oxide-based coatings of 3D-printed polycaprolactone (PCL) implants: graphene oxide (–GO), carboxylated GO (–GO–COOH) and reduced GO (–rGO). The effects of surface features and CL were evaluated in vitro using bone marrow-derived mesenchymal stromal cells (hBM-MSC). Our results showed that the CL formed on negatively charged PCL–GO–COOH and PCL–rGO scaffolds reduced cell adhesion, while simultaneously enhancing cell cluster formation and proliferation by a fivefold increase. The quantification of bone mineralized matrix highlighted that CL on both PCL–GO–COOH and PCL–rGO coatings sustained the osteogenic potential of these two types of GO. The analysis of CL components adsorbed on the scaffolds revealed that the PCL–GO–COOH and PCL–rGO coatings tend to entrap specific patterns of serum proteins (e.g. anti-adhesive and osteogenic modulators) and ions (carbonate and phosphate), suggesting a correlation between these enriched components and the observed biological outcomes of conditioned scaffolds. Lastly, PCL–rGO coatings maintained unique antibacterial properties after in vitro simulated CL formation, representing a suitable promising strategy to improve bone grafting capable of shaping CL formation while preserving the favorable osteoinductive properties of scaffolds.
Blood, a ubiquitous and fundamental carbohydrate material composed of plasma, red blood cells, white blood cells,
and platelets, has been playing an important role in biology, life science, history, and religious studies, while graphene has garnered significant attention due to its exceptional properties and extensive range of potential applications. Achieving environmentally friendly, cost-effective growth using hybrid precursors and obtaining high-quality graphene through a straightforward chemical vapor deposition process have been traditionally considered mutually exclusive. This study demonstrates that we can produce high-quality graphene domains with controlled thickness through a one-step growth process at atmospheric pressure using blood as a precursor. Raman spectroscopy confirms the uniformity of the blood-grown graphene films, and observing the half-integer quantum Hall effect in the measured devices highlights their outstanding electronic properties. This unprecedented approach opens possibilities for blood application, facilitating an unconventional route for graphene growth applications.
The biology of myeloid-derived suppressor cells (MDSCs) can be modified when grown on reduced graphene oxide (rGO) films. A higher oxidation state and roughness of rGO deplete MDSC activity by impacting on cell viability.
Two-dimensional (2D) materials have attracted tremendous interest ever since the isolation of atomically thin sheets of graphene in 2004 due to the specific and versatile properties of these materials. However, the increasing production and use of 2D materials necessitate a thorough evaluation of the potential impact on human health and the environment. Furthermore, harmonized test protocols are needed with which to assess the safety of 2D materials. The Graphene Flagship project (2013–2023), funded by the European Commission, addressed the identification of the possible hazard of graphene-based materials as well as emerging 2D materials including transition metal dichalcogenides, hexagonal boron nitride, and others. Additionally, so-called green chemistry approaches were explored to achieve the goal of a safe and sustainable production and use of this fascinating family of nanomaterials. The present review provides a compact survey of the findings and the lessons learned in the Graphene Flagship.
Carbon nanomaterials (CNMs), including carbon nanotubes (CNTs), graphene, and nanodiamonds (NDs), have shown great promise in detecting and treating numerous cancers, including kidney cancer. CNMs can increase the sensitivity of diagnostic techniques for better kidney cancer identification and surveillance. They enable targeted medicine delivery specifically to tumour locations, with little effect on healthy tissue. Because of their unique chemical and physical characteristics, they can avoid the body's defence mechanisms, making it easier to accumulate where tumours exist. Consequently, CNMs provide more effective drug delivery to kidney cancer cells. It also helps in improving the efficacy of treatment. This review explores the potential of several CNMs in improving therapeutic strategies for kidney cancer. We briefly covered the physicochemical properties and therapeutic applications of CNMs. Additionally, we discussed how structural modifications in CNMs enhance their precision in treating renal cancer. A thorough overview of CNM-based gene, peptide, and drug delivery strategies for the treatment of renal cancer is presented in this review. It covers information on other CNM-based therapeutic approaches, such as hyperthermia, photodynamic therapy, and photoacoustic therapy. Also, the interactions of CNMs with the tumour microenvironment (TME) are explored, including modulation of the immune response, regulation of tumour hypoxia, interactions between CNMs and TME cells, effects of TME pH on CNMs, and more. Finally, potential side effects of CNMs, such as toxicity, bio corona formation, enzymatic degradation, and biocompatibility, are also discussed.
Two-dimensional (2D) materials, such as MXenes and graphene, are tested as catalysts, sensors, and nanomedicine, but their safety and biocompatibility are poorly understood. In this study, Ti3C2 MXenes (single-layer and multilayer) and graphene oxide are prepared to evaluate their inflammatory potential in vivo and in vitro using the read-across approach. Pulmonary exposure of the test materials in mice led to significant dose-dependent increases in several toxicity endpoints of bronchoalveolar lavage fluid. The correlation of the oxidative potential of the test materials and the toxicity endpoints shows that both the intrinsic and intracellular oxidative potentials of 2D materials are key factors for the inflammation and damage of the plasma membrane. Scanning electron microscopy and in vitro studies using the differentiated THP-1 macrophage-like cells support this finding. Furthermore, the toxicity of test items is reduced when treated with N-acetyl cysteine, a scavenger for reactive oxygen species; this finding further supports the oxidative stress paradigm proposed in this study. This oxidative stress paradigm may apply to 2D materials in general and improve the understanding the toxicity mechanism of such materials, which would facilitate the design of safer functional 2D materials.
Biocompatible adsorbents play an essential role in hemoperfusion. Nevertheless, there are no hemoperfusion adsorbents that can simultaneously remove small and medium toxins, including bilirubin, urea, phosphor, heavy metals, and antibiotics. This bottleneck significantly impedes the miniaturization and portability of hemoperfusion materials and devices. Herein, a biocompatible protein‐polysaccharide complex is reported that exhibits “multi‐in‐one” removal efficacy for liver and kidney metabolism wastes, toxic metal ions, and antibiotics. Through electrostatic interactions and polysaccharide‐mediated coacervation, adsorbents can be prepared by simply mixing lysozyme (LZ) and sodium alginate (SA) together in seconds. The LZ/SA absorbent presented high adsorption capacities for bilirubin, urea, and Hg²⁺ of up to 468, 331, and 497 mg g⁻¹, respectively, and the excellent anti‐protein adsorption endowed LZ/SA with a record‐high adsorption capacity for bilirubin in the interference of serum albumin to simulate the physiological environment. The LZ/SA adsorbent also has effective adsorption capacity for heavy metals (Pb²⁺, Cu²⁺, Cr³⁺, and Cd²⁺) and multiple antibiotics (terramycin, tetracycline, enrofloxacin, norfloxacin, roxithromycin, erythromycin, sulfapyrimidine, and sulfamethoxazole). Various adsorption functional groups exposed on the adsorbent surface significantly contribute to the excellent adsorption capacity. This fully bio‐derived protein/alginate‐based hemoperfusion adsorbent has great application prospects in the treatment of blood‐related diseases.
Reduction of renal function, such as creatinine adsorption is one of the most common and dangerous diseases. Dedicated to this issue, developing high-performance, sustainable, and bio-compatible adsorbing materials is still challenging. Herein, barium alginate (BA) and BA containing few-layer graphene (FLG/BA) beads were synthesized in water from sodium alginate, also acting as bio-surfactant in in-situ exfoliation of graphite to FLG. The physicochemical characteristics of the beads demonstrated an excess of barium chloride used as a cross-linker. The efficiency and sorption capacity (Qe) of creatinine removal increase with processing duration reaching 82.1, 99.5 %, and 68.4, 82.9 mg·g-1 for BA and FLG/BA, respectively. The thermodynamic parameters detect the enthalpy change (ΔH°) of about -24.29 and -36.11 kJ·mol-1 and the entropy change (ΔS°) of around -69.24 and -79.46 kJ·mol-1 for BA and FLG/BA, respectively. During the reusability test, the removal efficiency decreases from the optimal first cycle to 69.1 and to 88.3 % in the sixth cycle for BA and FLG/BA, revealing superior stability of FLG/BA. The MD calculations confirm a higher adsorption capacity of FLG/BA composite compared to BA alone, clearly confirming a strong structure-property relation.
The Coronavirus Disease 2019 (COVID-19) pandemic has led to collaboration between nanotechnology scientists, industry stakeholders, and clinicians to develop solutions for diagnostics, prevention, and treatment of Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) infections. Nanomaterials, including carbon-based materials (CBM) such as graphene and carbon nanotubes, have been studied for their potential in viral research. CBM unique effects on microorganisms, immune interaction, and sensitivity in diagnostics have made them a promising subject of SARS-CoV-2 research. This review discusses the interaction of CBM with SARS-CoV-2 and their applicability, including CBM physical and chemical properties, the known interactions between CBM and viral components, and the proposed prevention, treatment, and diagnostics uses.
Procalcitonin (PCT) is one of the core biomarkers for the body’s systemic inflammatory response that is expressed uniformly in several organ tissues. PCT concentration levels rise significantly reaching the highest peak within 48 h after the bacterial infection starts. In this study, we evaluated different polymer-functionalized carbon nanotubes (CNTs) to enhance the performance of a sensing platform for the rapid detection of PCT over a large physiological range. CNTs were treated with polyethylene glycol (PEG), polyvinylpyrrolidone (PVP), and poly(L-arginine) P(Arg) to improve the nanostructures while promoting stability and reducing non-specific interaction. Different morphology analyses were performed to investigate the properties of the polymer-functionalized CNTs and compared them with the pristine material. Screen-printed carbon electrode (SPCE) sensors were modified with the three proposed nanocomposites and electrical characterization are performed. Afterward, the PCT antibody (100 ng/ml) is immobilized on the working electrodes before the electrochemical measurements with the analytes. Differential pulse voltammetry (DPV) responses were measured to quantify the PCT (27.43–20,000 pg/ml) in buffer solution and human plasma samples. CNT-PVP and CNT-P(Arg) electrodes presented a high correlation between the current peaks and the concentration of PCT. Subsequently, an interference study was carried out with inflammatory-related biomarkers that could cause a false positive response, the variation of the signal current was less than 3%. The SPCE devices also went through validation with the “gold-standard” laboratory method to verify the agreement with the proposed method. This study regarding the optimization of the CNTs is just the first part of our final target regarding a multiplexed point-of-care device using a small volume of the blood sample to provide a rapid diagnostic regarding inflammatory response.
Titanium (Ti) was an excellent medical metal material, but the lack of good antibacterial activity confined its further practical application. To solve this dilemma, a coating containing graphene oxide (GO) and copper (Cu) was prepared on the surface of Ti sheet (Ti/APS/GO/Cu). First, physical sterilization could be carried out through the sharp-edged sheet structure of GO. Second, the oxygen-containing functional group on the surface of GO and the released Cu2+ would generate reactive oxygen species for chemical sterilization. The synergistic effect of GO and Cu substantially enhanced the in vitro and in vivo antibacterial property of Ti sheet, thereby reducing bacterial-related inflammation. Quantitatively, the antibacterial rate of Ti/APS/GO/Cu against E. coli or S. aureus reached over 99%. Besides, Ti/APS/GO/Cu showed excellent biocompatibility and no toxicity to cell. Such work developed multiple sterilization avenues to design non-antibiotic, safe and efficient antibacterial implant material for the biomedical domain.
A semi-automated diffusion-dialysis purification procedure is proposed for the preparation of uncontaminated graphene oxide (GO) aqueous dispersions. The purification process is integrated with analytical-signal processing to control the purification degree online by several channels: oxidation-reduction potential, conductivity, and absorbance. This approach reduces the amounts of reagents for chemical treatment during dialysis. The total transition metal (Mn and Ti) content was reduced to a sub-ppb level (assessed by slurry nebulization in inductively coupled plasma optical atomic emission spectroscopy). Purified aqueous GO samples possess good stability for about a year with a zeta-potential of ca. −40 mV and a lateral size of ca. sub-µm. Purified GO samples showed increased antioxidant properties (up to five times compared to initial samples according to chemiluminometry by superoxide-radical (O2−) generated in situ from xanthine and xanthine oxidase with the lucigenin probe) and significantly decreased peroxidase-like activity (assessed by the H2O2–L-012 system).
This work start after seeing an recent open letter for transparency related production an quality control technique of mRNA vaccine first signed by Tarro G, Luisetto M and Monsellato ML and an editorial recognized by IMA Marijnskaya academy: graphene and derivates: physico-chemical and toxicology properties in the mRNA vaccine manufacturing strategy Needed specific proof of absence for the regulatory aspects (accepted for publication). Other relevant evidences comes from the work of Giovannini et al related DARKFIED microscope assay of the blood of 1086 symptomatic subjects after vaccination with two types of mRNA vaccine of great interest on this field the work of P Campra and Young RO, Young Me Lee or Ki-Yeob J. Aim of this work is to 2 Research Article | Mauro L, et al. Genesis J Surg Med. 2022, 1(2)-6. biotechnological application like mRNA vaccine. After a review part an experimental hypotesys project will be submitted to the researcher to produce a global conclusion. The recent evidences published in last period induced the idea to more deeply study this properties for The clinico-toxicological aspects involved.
This work start after seeing an recent open letter for transparency related production an quality control technique of mRNA vaccine first signed by Tarro G, Luisetto M and Monsellato ML and an editorial recognized by IMA Marijnskaya academy: graphene and derivates: physico-chemical and toxicology properties in the mRNA vaccine manufacturing strategy Needed specific proof of absence for the regulatory aspects (accepted for publication). Other relevant evidences comes from the work of Giovannini et al related DARKFIED microscope assay of the blood of 1086 symptomatic subjects after vaccination with two types of mRNA vaccine of great interest on this field the work of P Campra and Young RO, Young Me Lee or Ki-Yeob J. Aim of this work is to 2 Research Article | Mauro L, et al. Genesis J Surg Med. 2022, 1(2)-6. biotechnological application like mRNA vaccine. After a review part an experimental hypotesys project will be submitted to the researcher to produce a global conclusion. The recent evidences published in last period induced the idea to more deeply study this properties for The clinico-toxicological aspects involved.
Leveraging the phononic sensitivity and scalability of nano‐biointerfaces has accelerated the growth of unique and versatile biosensors. Graphene has the properties of a near‐ideal signal transducer, due to the strong coupling between its interfacial and phononic properties. This enables sensitive yet quick detection of surface interactions on graphene via Raman spectral analysis. The Raman‐active vibrational bands of graphene are demonstrated to be sensitive to structural, electrical, and interfacial modifications. This sensitivity is attributed to graphene's electron–phonon coupling and high quantum capacitance. The fundamental understanding of graphene phonons is crucial for developing reliable platforms for disease and infectious agent detection. This review provides a mechanistic explanation of these phenomena at the interface between graphene and various biosystems (including cancerous, bacterial, viral, and biophysical specimens) to set the foundation for next‐generation chemeo‐phononic medical devices. The interface of ultrasensitive graphene with various biological specimens enables the Raman‐active transducer to monitor the surface potential in real time. The underlying fundamentals of the phononic graphene transducer are explained, and the impact of the relevant works is described. Graphene's strong interfacial doping characteristic coupled with its phononics produces a robust chemeo‐phononic transducer that presents numerous novel biomedical applications.
Background
The effect of bionanointeractions on graphene-biomolecule nanohybrids is of great interest, since external influences on their structural and surface properties can significantly affect their biological activity.
Introduction
The effects of the fatty acid binding with shungite carbon (ShC) nanoparticles on the stability of aqueous dispersions of ShC and the oxidation state of ShC (oxygen-containing groups) were studied using linoleic acid (LA) as an example
Method
The size and surface charge (ζ -potential) of the ShC-LA associates formed at various LA concentrations in the dispersion were estimated using the dynamic light scattering method and the ultraviolet (UV) absorption spectra of dispersions were taken.
Result
The negative ShC charge becomes less negative upon LA binding, depending on LA concentration. The size of ShC upon functionalization by LA molecules does not depend on LA concentration, suggesting the predominance of surface rearrangement of NPs, rather than a change in their global structure. ShC - LA interaction is accompanied by an increase in absorption in the UV spectrum region of conjugated С=С bonds, the reduction of С=О groups, sp2 hybridization and bonds in the plane of graphene fragments, the basic structural units of ShC.
Conclusion
The results are interpreted in terms of the surface structural effects of LA on ShC that affect variations of the colloid and redox characteristics of ShC in aqueous dispersion.
Nanotechnology has a great potential to revolutionize the landscape of medicine, but an inadequate understanding of the nanomaterial-biological (nano-bio) interface hampers its ultimate clinical translation. Surface attachment of biomolecules provides a new biological identity of nanoparticles that plays a crucial role in vivo as it can activate the immune system triggering inflammatory responses, clearance from the body, and cellular toxicity. In this review, we summarize and critically analyze progress in understanding the relationship between the biological identity of nanoparticles and immune system activation. Accordingly, we discuss the implications of biomolecular corona on nanotoxicity, immune safety, and biocompatibility. We also highlight a perspective on engineering the biological identity of nanoparticles for modulating immunological responses.
Ultrasound has important applications, predominantly in the field of diagnostic imaging. Presently, colloidal systems such as microbubbles, phase-change emulsion droplets and particle systems with acoustic properties and multiresponsiveness are being developed to address typical issues faced when using commercial ultrasound contrast agents, and to extend the utility of such systems to targeted drug delivery and multimodal imaging. Current technologies and increasing research data on the chemistry, physics and materials science of new colloidal systems are also leading to the development of more complex, novel and application-specific colloidal assemblies with ultrasound contrast enhancement and other properties, which could be beneficial for multiple biomedical applications, especially imaging-guided treatments. In this article, we review recent developments in new colloids with applications that use ultrasound contrast enhancement. This work also highlights the emergence of colloidal materials fabricated from or modified with biologically derived and bio-inspired materials, particularly in the form of biopolymers and biomembranes. Challenges, limitations, potential developments and future directions of these next-generation colloidal systems are also presented and discussed.
Graphene and its derivatives possess some intriguing properties, which generates tremendous interests in various fields, including biomedicine. The biomedical applications of graphene-based nanomaterials have attracted great interests over the last decade, and several groups have started working on this field around the globe. Because of the excellent biocompatibility, solubility and selectivity, graphene and its derivatives have shown great potential as biosensing and bio-imaging materials. Also, due to some unique physicochemical properties of graphene and its derivatives, such as large surface
area, high purity, good bio-functionalizability, easy solubility, high drug loading capacity, capability of easy cell membrane penetration, etc., graphene-based nanomaterials become promising candidates for bio-delivery carriers. Besides, graphene and its derivatives have also shown interesting applications in the fields of cell-culture, cell-growth and tissue engineering. In this article, a comprehensive review on the applications of graphene and its derivatives as biomedical materials has been presented. The unique properties of graphene and its derivatives (such as graphene oxide, reduced graphene oxide, graphane, graphone, graphyne, graphdiyne, fluorographene and their doped versions) have been discussed, followed by discussions on the recent efforts on the applications of graphene and its derivatives in biosensing, bio-imaging, drug delivery and therapy, cell culture, tissue engineering and cell growth. Also, the challenges involved in the use of graphene and its derivatives as biomedical materials are discussed briefly, followed by the future perspectives of the use of graphene-based nanomaterials in bioapplications. The review will provide an outlook to the applications of graphene and its derivatives, and may open up new horizons to inspire broader interests across various disciplines.
Graphene-based nanomaterials (GBNs) have attracted increasing interests of the scientific community due to their unique physicochemical properties and their applications in biotechnology, biomedicine, bioengineering, disease diagnosis and therapy. Although a large amount of researches have been conducted on these novel nanomaterials, limited comprehensive reviews are published on their biomedical applications and potential environmental and human health effects. The present research aimed at addressing this knowledge gap by examining and discussing: (1) the history, synthesis, structural properties and recent developments of GBNs for biomedical applications; (2) GBNs uses as therapeutics, drug/gene delivery and antibacterial materials; (3) GBNs applications in tissue engineering and in research as biosensors and bioimaging materials; and (4) GBNs potential environmental effects and human health risks. It also discussed the perspectives and challenges associated with the biomedical applications of GBNs. Open image in new window
Methicillin-resistant Staphylococcus aureus (MRSA) is responsible for serious hospital infections worldwide and represents a global public health problem. Curcumin, the major constituent of turmeric, is effective against MRSA but only at cytotoxic concentrations or in combination with antibiotics. The major issue in curcumin-based therapies is the poor solubility of this hydrophobic compound and the cytotoxicity at high doses. In this paper, we describe the efficacy of a composite nanoparticle made of curcumin (CU) and graphene oxide (GO), hereafter GOCU, in MRSA infection treatment. GO is a nanomaterial with a large surface area and high drug-loading capacity. GO has also antibacterial properties due mainly to a mechanical cutting of the bacterial membranes. For this physical mechanism of action, microorganisms are unlikely to develop resistance against this nanomaterial. In this work, we report the capacity of GO to support and stabilize curcumin molecules in a water environment and we demonstrate the efficacy of GOCU against MRSA at a concentration below 2 µg ml-1. Further, GOCU displays low toxicity on fibroblasts cells and avoids haemolysis of red blood cells. Our results indicate that GOCU is a promising nanomaterial against antibiotic-resistant MRSA.
The systematic study of nanoparticle-biological interactions requires particles to be reproducibly dispersed in relevant fluids along with further development in the identification of biologically relevant structural details at the materials-biology interface. Here, we develop a biocompatible long-term colloidally stable water dispersion of few-layered graphene nanoflakes in the biological exposure medium in which it will be studied. We also report the study of the orientation and functionality of key proteins of interest in the biolayer (corona) that are believed to mediate most of the early biological interactions. The evidence accumulated shows that graphene nanoflakes are rich in effective apolipoprotein A-I presentation, and we are able to map specific functional epitopes located in the C-terminal portion that are known to mediate the binding of high-density lipoprotein to binding sites in receptors that are abundant in the liver. This could suggest a way of connecting the materials' properties to the biological outcomes.
The successful applications of graphene nanomaterials in nanobiotechnology and medicine as well as their effective translation into real clinical utility hinge significantly on a thorough understanding of their nanotoxicological profile. Of all aspects of biocompatibility, the hemocompatibility of graphene nanomaterials with different blood constituents in the circulatory system is one of the most important elements that needs to be well elucidated. Once administered into biological systems, graphene nanomaterials may inevitably come into contact with the surrounding plasma proteins and blood cells. Crucially, the interactions between these hematological entities and graphene nanomaterials will influence the overall efficacy of their biomedical applications. As such, a comprehensive understanding of the hemotoxicity of graphene nanomaterials is critically important. This review presents an up-to-date elucidation of the hemotoxicity of graphene nanomaterials through their interactions with blood proteins and cells, as well as offers some perspectives on the current challenges, opportunities, and future development of this important field. Copyright © Materials Research Society 2017 This is an Open Access article, distributed under the terms of the Creative Commons Attribution licence (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted re-use, distribution, and reproduction in any medium, provided the original work is properly cited.
Carbon nanomaterials are some of the most versatile nanomaterials. Along with increasing explorations into their utilization in a plethora of biological and biomedical applications, there have been emerging interests and needs in understanding the molecular hemocompatibility of these engineered nanomaterials when coming into contact with blood. Here, we evaluate the nano-bio interactions of one-dimensional (1D) and two-dimensional (2D) carbon nanomaterials with blood plasma proteins. Different facets of the nanomaterial–protein interactions, specifically, the adsorption, equilibrium binding and conformational stability of proteins upon association with carbon nanomaterials are established, based on the quantification of various parameters, such as association constant, binding cooperativity and protein secondary structural change. In light of our data, we demonstrate that the carbon nanomaterial–plasma protein interactions may be significantly influenced by the density of the oxygenated functionalities of the nanomaterials and to a certain extent, their dimensionality and surface area. This work offers a broad insight into the nano-bio interactions between carbon nanomaterials and blood plasma proteins and provides a strong basis for the design and use of 1D and 2D carbon nanomaterials for a wide variety of bioapplications.
The graphene family has captured the interest and the imagination of an increasing number of scientists working in different fields, ranging from composites to flexible electronics. In the area of biomedical applications, graphene is especially involved in drug delivery, biosensing and tissue engineering, with strong contributions to the whole nanomedicine area. Besides the interesting results obtained so far and the evident success, there are still many problems to solve, on the way to the manufacturing of biomedical devices, including the lack of standardization in the production of the graphene family members. Control of lateral size, aggregation state (single vs. few layers) and oxidation state (unmodified graphene vs. oxidized graphenes) is essential for the translation of this material into clinical assays. In this Tutorial Review we critically describe the latest developments of the graphene family materials into the biomedical field. We analyze graphene-based devices starting from graphene synthetic strategies, functionalization and processibility protocols up to the final in vitro and in vivo applications. We also address the toxicological impact and the limitations in translating graphene materials into advanced clinical tools. Finally, new trends and guidelines for future developments are presented.
For the first time a critical analysis of the influence that four different graphene oxide reduction methods have on the electrochemical properties of the resulting reduced graphene oxides (RGOs) is reported. Starting from the same graphene oxide, chemical (CRGO), hydrothermal (hTRGO), electrochemical (ERGO), and thermal (TRGO) reduced graphene oxide were produced. The materials were fully characterized and the topography and electroactivity of the resulting glassy carbon modified electrodes were also evaluated. An oligonucleotide molecule was used as a model of DNA electrochemical biosensing. The results allow for the conclusion that TRGO produced the RGOs with the best electrochemical performance for oligonucleotide electroanalysis. A clear shift in the guanine oxidation peak potential to lower values (~0.100 V) and an almost two-fold increase in the current intensity were observed compared with the other RGOs. The electrocatalytic effect has a multifactorial explanation because the TRGO was the material that presented a higher polydispersity and lower sheet size, thus exposing a larger quantity of defects to the electrode surface, which produces larger physical and electrochemical areas.
Graphene-based materials (GBMs) are emerging as attractive materials for biomedical applications. Understanding how these materials are perceived by and interact with the immune system is of fundamental importance. Phagocytosis is a major mechanism deployed by the immune system to remove pathogens, particles, and cellular debris. Here, we discuss recent studies on the interactions of GBMs with different phagocytic cells, including macrophages, neutrophils, and dendritic cells. The importance of assessing GBMs for endotoxin contamination is discussed as this may skew results. We also explore the role of the bio-corona for interactions of GBMs with immune cells. Finally, we highlight recent evidence for direct plasma membrane interactions of GBMs.
The development of new pharmacological strategies that evade bacterial resistance has become a compelling worldwide challenge. Graphene oxide (GO) can represent the nanotechnology answer being economical and easy to produce and to degrade and having multitarget specificity against bacteria. Several groups tried to define the interaction between GO sheets and human pathogens. Unfortunately, controversial results from inhibition to bacterial growth enhancement have been reported. The main difference among all experimental evidence relies on the environmental conditions adopted to study the bacteria-GO interaction. Indeed GO, stable in deionized water, undergoes a rapid and salt-specific DLVO-like aggregation that influences antimicrobial effects. Considering this phenomenon, the interaction of bacteria with GO aggregates having different sizes, morphologies, and surface potential can create a complex scenario that explains the contrasting results reported so far. In this article, we demonstrate that by modulating the GO stability in solution, the antibacterial or growth enhancement effect can be controlled on S. aureus and E. coli. GO at low concentration cuts microorganism membranes and at high concentration forms complexes with pathogens and inhibits or enhances bacterial growth in a surface potential-dependent manner. With the framework defined in this study, the clinical application of GO gets closer, and controversial results in literature can be explained.
Standing out as the new wonder bidimensional material, graphene oxide (GO) has aroused an exceptional interest in biomedical research by holding promise for being the antibacterial of future. First, GO possesses a specific interaction with microorganisms combined with a mild toxicity for human cells. Additionally, its antibacterial action seems to be directed to multiple targets in pathogens, causing both membranes mechanical injury and oxidative stress. Lastly, compared to other carbon materials, GO has easy and low-cost processing and is environment-friendly. This remarkable specificity and multi-targeting antibacterial activity come at a time when antibiotic resistance represents the major health challenge. Unfortunately, a comprehensive framework to understand how to effectively utilize this material against microorganisms is still lacking. In the last decade, several groups tried to define the mechanisms of interaction between GO flakes and pathogens but conflicting results have been reported. This review is focused on all the contradictions of GO antimicrobial properties in solution. Flake size, incubation protocol, time of exposure and species considered are examples of factors influencing results. These parameters will be summarized and analyzed with the aim of defining the causes of contradictions, to allow fast GO clinical application.
The application of graphene oxide (GO) as a potential vaccine adjuvant has recently attracted considerable attention. However, appropriate surface functionalization of GO is crucial to improve its biocompatibility and enhance its adjuvant activity. In this study, we developed a simple method to prepare chitosan (CS)-functionalized GO (GO-CS) and further investigated its potential as a nanoadjuvant. Compared with GO, GO-CS possessed considerably smaller size, positive surface charge, and better thermal stability. The functionalization of GO with CS was effective in decreasing the non-specific protein adsorption and improving its biocompatibility. Furthermore, GO-CS significantly activated RAW264.7 cells and stimulated more cytokines for mediating cellular immune response, which was mainly due to the synergistic immunostimulatory effect of both GO and CS. GO-CS exhibits strong potential as a safe nanoadjuvant for vaccines and immunotherapy.
Engineered nanomaterials promise to transform medicine at the bio–nano interface. However, it is important to elucidate how synthetic nanomaterials interact with critical biological systems before such products can be safely utilized in humans. Past evidence suggests that polyethylene glycol-functionalized (PEGylated) nanomaterials are largely biocompatible and elicit less dramatic immune responses than their pristine counterparts. We here report results that contradict these findings. We find that PEGylated graphene oxide nanosheets (nGO-PEGs) stimulate potent cytokine responses in peritoneal macrophages, despite not being internalized. Atomistic molecular dynamics simulations support a mechanism by which nGO-PEGs preferentially adsorb onto and/or partially insert into cell membranes, thereby amplifying interactions with stimulatory surface receptors. Further experiments demonstrate that nGO-PEG indeed provokes cytokine secretion by enhancing integrin β8-related signalling pathways. The present results inform that surface passivation does not always prevent immunological reactions to 2D nanomaterials but also suggest applications for PEGylated nanomaterials wherein immune stimulation is desired.
We unravel the role of flake dimensionality on the lithiation/de-lithiation processes and electrochemical performance of anodes based on few-(FLG) and multi-layer graphene (MLG) flakes prepared by liquid phase exfoliation (LPE) of pristine graphite. The flakes are sorted by lateral size (from 380 to 75 nm) and thickness from 20 (MLG) to 2 nm (FLG) exploiting a sedimentation-based separation in centrifugal field and, finally, deposited onto Cu disks for the realization of four binder-free anodes. The electrochemical results show that decreasing lateral size and thickness leads to an increase of the initial specific capacity from ≈590 to ≈1270mAhg−1. However, an increasing irreversible capacity is also associated to the reduction of flakes’ size. We find, in addition, that the preferential Li ions storage by adsorption rather than intercalation in small lateral size (<100 nm) FLG flakes has a detrimental effect on the average de-lithiation voltage, resulting on low voltage efficiency of these anodes. We believe that the results reported in this work, provide the guidelines for the practical exploitation of graphene-based electrodes.
Antibacterial surfaces have an enormous economic and social impact on the worldwide technological fight against diseases. However, bacteria develop resistance and coatings are often not uniform and not stable in time. The challenge is finding an antibacterial coating that is biocompatible, cost-effective, not toxic, and spreadable over large and irregular surfaces. Here we demonstrate an antibacterial cloak by laser printing of graphene oxide hydrogels mimicking the Cancer Pagurus carapace. We observe up to 90% reduction of bacteria cells. This cloak exploits natural surface patterns evolved to resist to microorganisms infection, and the antimicrobial efficacy of graphene oxide. Cell integrity analysis by scanning electron microscopy and nucleic acids release show bacteriostatic and bactericidal effect. Nucleic acids release demonstrates microorganism cutting, and microscopy reveals cells wrapped by the laser treated gel. A theoretical active matter model confirms our findings. The employment of biomimetic graphene oxide gels opens unique possibilities to decrease infections in biomedical applications and chirurgical equipment; our antibiotic-free approach, based on the geometric reduction of microbial adhesion and the mechanical action of Graphene Oxide sheets, is potentially not affected by bacterial resistance.
Pancreatic cancer is a very aggressive malignancy that is often diagnosed in the advanced stages, with the implication that long-term survivors are extremely rare. Thus, developing new methods for the early detection of pancreatic cancer is an urgent task for current research. To date, nanotechnology offers unprecedented opportunities for cancer therapeutics and diagnosis. The aim of this study is the development of a new pancreatic cancer diagnostic technology based on the exploitation of the nano-bio-interactions between nanoparticles and blood samples. In this study, blood samples from 20 pancreatic cancer patients and 5 patients without malignancy were allowed to interact with designed lipid nanoparticles, leading to the formation of a hard "protein corona" at the nanoparticle surface. After isolation, the protein patterns were characterized by sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS PAGE). We found that the protein corona of pancreatic cancer patients was much more enriched than that of healthy individuals. Statistical analysis of SDS-PAGE results allowed us to discriminate between healthy and pancreatic cancer patients with a total discriminate correctness rate of 88%.
Breast cancer (BC) results in ~40,000 deaths each year in the United States and even among survivors treatment of the disease may have devastating consequences, including increased risk for heart disease and cognitive impairment resulting from the toxic effects of chemotherapy. Aptamer-mediated drug delivery can contribute to improved treatment outcomes through the selective delivery of chemotherapy to BC cells, provided suitable cancer-specific antigens can be identified. We report here the use of capillary electrophoresis in conjunction with next generation sequencing to develop the first vitronectin (VN) binding aptamer (VBA-01; Kd 405 nmol/l, the first aptamer to vitronectin (VN; Kd = 405 nmol/l) , a protein that plays an important role in wound healing and that is present at elevated levels in BC tissue and in the blood of BC patients relative to the corresponding nonmalignant tissues. We used VBA-01 to develop DVBA-01, a dimeric aptamer complex, and conjugated doxorubicin (Dox) to DVBA-01 (7:1 ratio) using pH-sensitive, covalent linkages. Dox conjugation enhanced the thermal stability of the complex (60.2 versus 46.5°C) and did not decrease affinity for the VN target. The resulting DVBA-01-Dox complex displayed increased cytotoxicity to MDA-MB-231 BC cells that were cultured on plasticware coated with VN (1.8 × 10⁻⁶mol/l) relative to uncoated plates (2.4 × 10⁻⁶ mol/l), or plates coated with the related protein fibronectin (2.1 × 10⁻⁶ mol/l). The VBA-01 aptamer was evaluated for binding to human BC tissue using immunohistochemistry and displayed tissue specific binding and apparent association with BC cells. In contrast, a monoclonal antibody that preferentially binds to multimeric VN primarily stained extracellular matrix and vessel walls of BC tissue. Our results indicate a strong potential for using VN-targeting aptamers to improve drug delivery to treat BC.
Due to their unique physicochemical properties, graphene-family nanomaterials (GFNs) are widely used in many fields, especially in biomedical applications. Currently, many studies have investigated the biocompatibility and toxicity of GFNs in vivo and in intro. Generally, GFNs may exert different degrees of toxicity in animals or cell models by following with different administration routes and penetrating through physiological barriers, subsequently being distributed in tissues or located in cells, eventually being excreted out of the bodies. This review collects studies on the toxic effects of GFNs in several organs and cell models. We also point out that various factors determine the toxicity of GFNs including the lateral size, surface structure, functionalization, charge, impurities, aggregations, and corona effect ect. In addition, several typical mechanisms underlying GFN toxicity have been revealed, for instance, physical destruction, oxidative stress, DNA damage, inflammatory response, apoptosis, autophagy, and necrosis. In these mechanisms, (toll-like receptors-) TLR-, transforming growth factor β- (TGF-β-) and tumor necrosis factor-alpha (TNF-α) dependent-pathways are involved in the signalling pathway network, and oxidative stress plays a crucial role in these pathways. In this review, we summarize the available information on regulating factors and the mechanisms of GFNs toxicity, and propose some challenges and suggestions for further investigations of GFNs, with the aim of completing the toxicology mechanisms, and providing suggestions to improve the biological safety of GFNs and facilitate their wide application.
Carbon-based functional nanomaterials have attracted immense scientific interest from many disciplines and, due to their extraordinary properties, have offered tremendous potential in a diverse range of applications. Among the different carbon nanomaterials, graphene is one of the newest and is considered the most important. Graphene, a monolayer material composed of sp(2)-hybridized carbon atoms hexagonally arranged in a two-dimensional structure, can be easily functionalized by chemical modification. Functionalized graphene and its derivatives have been used in diverse nano-biotechnological applications, such as in environmental engineering, biomedicine, and biotechnology. However, the prospective use of graphene-related materials in a biological context requires a detailed comprehension of these materials, which is essential for expanding their biomedical applications in the future. In recent years, the number of biological studies involving graphene-related nanomaterials has rapidly increased. These studies have documented the effects of the biological interactions between graphene-related materials and different organizational levels of living systems, ranging from biomolecules to animals. In the present review, we will summarize the recent progress in understanding mainly the interactions between graphene and cells. The impact of graphene on intracellular components, and especially the uptake and transport of graphene by cells, will be discussed in detail.
Background
We have previously demonstrated that reduced graphene oxide (rGO) administered intravenously in rats was detected inside the hippocampus after downregulation of the tight and adherens junction proteins of the blood–brain barrier. While down-regulators of junctional proteins could be useful tools for drug delivery through the paracellular pathway, concerns over toxicity must be investigated before clinical application. Herein, our purpose was to trace whether the rGO inside the hippocampus triggered toxic alterations in this brain region and in target organs (blood, liver and kidney) of rats at various time points (15 min, 1, 3 h and 7 days).
Results
The assessed rGO-treated rats (7 mg/kg) were clinically indistinguishable from controls at all the time points. Hematological, histopathological (neurons and astrocytes markers), biochemical (nephrotoxicity and hepatotoxicity assessment) and genotoxicological based tests showed that systemic rGO single injection seemed to produce minimal toxicological effects at the time points assessed. Relative to control, the only change was a decrease in the blood urea nitrogen level 3 h post-treatment and increases in superoxide dismutase activity 1 h and 7 days post-treatment. While no alteration in leukocyte parameters was detected between control and rGO-treated animals, time-dependent leukocytosis (rGO-1 h versus rGO-3 h) and leukopenia (rGO-3 h versus rGO-7 days) was observed intra-treated groups. Nevertheless, no inflammatory response was induced in serum and hippocampus at any time.
Conclusions
The toxic effects seemed to be peripheral and transitory in the short-term analysis after systemic administration of rGO. The effects were self-limited and non-significant even at 7 days post-rGO administration.
Background
The blood–brain barrier (BBB) is a complex physical and functional barrier protecting the central nervous system from physical and chemical insults. Nevertheless, it also constitutes a barrier against therapeutics for treating neurological disorders. In this context, nanomaterial-based therapy provides a potential alternative for overcoming this problem. Graphene family has attracted significant interest in nanomedicine because their unique physicochemical properties make them amenable to applications in drug/gene delivery and neural interface.
Results
In this study, reduced graphene oxide (rGO) systemically-injected was found mainly located in the thalamus and hippocampus of rats. The entry of rGO involved a transitory decrease in the BBB paracellular tightness, as demonstrated at anatomical (Evans blue dye infusion), subcellular (transmission electron microscopy) and molecular (junctional protein expression) levels. Additionally, we examined the usefulness of matrix-assisted laser desorption/ionization (MALDI) mass spectrometry imaging (MSI) as a new imaging method for detecting the temporal distribution of nanomaterials throughout the brain.
Conclusions
rGO was able to be detected and monitored in the brain over time provided by a novel application for MALDI-MSI and could be a useful tool for treating a variety of brain disorders that are normally unresponsive to conventional treatment because of BBB impermeability.
Recent years have witnessed unprecedented increase in the use of nanoparticles in various sectors viz. electronics, catalysis, agriculture, textile, cosmetics, bio-medicine, packaging, house-holds and food-associated consumer products. This has led to the inevitable release of nanoparticles into the environment, which can have negative impact on living beings. Humans can also be exposed to these nanoparticles either intentionally or accidently. Nanoparticles can enter in the human body through food chain, inhalation, open wounds, drugs and intravenous injections etc. In majority of these cases, the nanoparticles may pass through the various cell layers, cell sap and finally enter into the blood. Upon interaction with biological fluid, nanoparticles come in close proximity particularly to the proteins present in the fluid. The assembly of proteins surrounding the nanoparticle's surface is called as protein corona and their complex is known as protein-nanoparticle complex. Formation of protein corona is a vibrant and driving process, which plays a pivotal role in the functioning of nanoparticles in biological systems. Moreover, due to interaction of proteins with nanoparticles, conformational changes may occur in the native structure of protein, which may lead to change in the functioning of proteins towards its cellular interaction. The present review provides in-depth knowledge about the formation of protein corona around nanoparticles and the factors regulating this process. Further, it discusses various techniques that can be used for the protein corona analysis and obtaining information about molecular consequences upon nanoparticle's exposure. Finally, the functional aspects of protein-nanoparticle complex have been discussed in detail. In-depth understanding of protein-nanoparticles complex can be instrumental to generate well-suited nanoparticles with desired surface characteristics in the way to biological application.
Graphene oxide (GO) has attracted significant interest as a template material for multiple applications due to its two-dimensional nature and established functionalization chemistries. However, for applications toward stem cell culture and differentiation, GO is often reduced to form reduced graphene oxide, resulting in a loss of oxygen content. Here, we induce a phase transformation in GO and demonstrate its benefits for enhanced stem cell culture and differentiation, while conserving the oxygen content. The transformation results in clustering of oxygen atoms on the GO surface, which greatly improves its ability toward substance adherence and results in enhanced differentiation of human mesenchymal stem cell (hMSCs) towards the osteogenic lineage. Moreover, the conjugating ability of modified GO strengthened, which was examined by auxiliary osteogenic growth peptide conjugation. Overall, our work demonstrates GO’s potential for stem cell applications, while maintaining its oxygen content, which could be enable further functionalization and fabrication of novel nano-bio interfaces.
Wafer-scale integration of reduced graphene oxide with H-terminated Si(1 1 1) surfaces has been accomplished by electrochemical reduction of a thin film of graphene oxide deposited onto Si by drop casting. Two reduction methods have been assayed and carried out in an acetonitrile solution. The initial deposit was subjected either to potential cycling in a 0.1 M TBAPF6/CH3CN solution at scan rates values of 20 mV s⁻¹ and 50 mV s⁻¹, or to a potentiostatic polarization at Eλ,c = −3 V for 450 s. The resulting interface has been characterized in its surface composition, morphology and electrochemical behavior by X-ray photoelectron spectroscopy, Raman spectroscopy, atomic force microscopy and electrochemical measurements. The results evidence that few-layer graphene deposits on H-Si(1 1 1) were obtained after reduction, and use of organic instead of aqueous medium led to a very limited surface oxidation of the Si substrate and a very low oxygen-to-carbon ratio. The described approach is fast, simple, economic, scalable and straightforward, as one reduction cycle is already effective in promoting the establishment of a graphene-Si interface. It avoids thermal treatments at high temperatures, use of aggressive chemicals and the presence of metal contaminants, and enables preservation of Si(1 1 1) surface from oxidation.
The fluorescence quenching property of graphene oxide (GO) has been newly demonstrated and applied for fluorescence imaging and biosensing. In this work, a new nanostructure was designed for effectively studying quenching ability of GO. The key element in this design is the fabrication of a layer of rigid and thickness-adjustable silica spacer for manipulating the distance between the GO and fluorophores. First, a silica core modified with organic dye molecules was prepared, followed by the formation of a silica shell with a tunable thickness. Afterwards, the GO was wrapped around silica nanoparticles based on the electrostatic interaction between negatively charged GO and positively charged silica. The quenching efficiency of GO to different dye molecules was studied at various spacer thicknesses and varying concentrations of GO. Fluorescence lifetime of fluorophores was measured to determine the quenching mechanism. We found that the quenching efficiency of GO was still around 30 % when the distance between dyes and GO was increased to more than 30 nm, which indicated the long-distance quenching ability of GO and confirmed previous theoretical calculation. The quenching mechanisms were proposed schematically based on our experimental results. We expected that the proposed nanostructure could act as a feasible model for studying GO quenching property and shed light on designing GO-based fluorescence sensing systems.
Neutrophils were previously shown to digest oxidized carbon nanotubes through a myeloperoxidase (MPO)-dependent mechanism, and graphene oxide (GO) was found to undergo degradation when incubated with purified MPO, but there are no studies to date showing degradation of GO by neutrophils. Here we produced endotoxin-free GO by a modified Hummers’ method and asked whether primary human neutrophils stimulated to produce neutrophil extracellular traps or activated to undergo degranulation are capable of digesting GO. Biodegradation was assessed using a range of techniques including Raman spectroscopy, transmission electron microscopy, atomic force microscopy, and mass spectrometry. GO sheets of differing lateral dimensions were effectively degraded by neutrophils. As the degradation products could have toxicological implications, we also evaluated the impact of degraded GO on the bronchial epithelial cell line BEAS-2B. MPO-degraded GO was found to be non-cytotoxic and did not elicit any DNA damage. Taken together, these studies have shown that neutrophils, key players of the immune system, can digest GO.
Many potential therapeutic compounds for brain diseases fail to reach their molecular targets due to the impermeability of the blood-brain barrier, limiting their clinical development. Nanotechnology-based approaches might improve compounds pharmacokinetics by enhancing binding to the cerebrovascular endothelium and translocation into the brain. Adsorption of apolipoprotein E4 onto polysorbate(®) 80-stabilized nanoparticles to produce a protein corona allows the specific targeting of cerebrovascular endothelium. This strategy increased nanoparticle translocation into brain parenchyma, and improved brain nanoparticle accumulation 3-fold compared to undecorated particles (119.8 vs 40.5 picomoles; p<0.01). Apolipoprotein-enhanced nanoparticles have high clinical translational potential and may improve the development of nanotechnology-based medicine for a variety of neurological diseases.
Following exposure to biological milieus (e.g. after systemic administration), nanoparticles (NPs) get covered by an outer biomolecular corona (BC) that defines many of their biological outcomes, such as the elicited immune response, biodistribution, and targeting capabilities. In spite of this, the BC role in regulating the cellular uptake and the subcellular trafficking properties of NPs has remained elusive. Here, we tackle this issue by employing multicomponent (MC) lipid NPs, human plasma (HP) and HeLa cells as models for nanoformulation, biological fluid, and target cell (respectively). By confocal fluorescence microscopy experiments and image correlation analyses, we quantitatively demonstrate that the BC promotes a neat switch of cell entry mechanism and subsequent intracellular trafficking, from macropinocytosis to clathrin-dependent endocytosis. Nano liquid chromatography tandem mass spectrometry identifies Apolipoproteins as the most abundant components of the BC tested here. Interestingly, this class of proteins target the LDL receptors, which are overexpressed in clathrin-enriched membrane domains. Our results highlight the crucial role of BC as an intrinsic trigger of specific NP-cell interactions and biological responses and set the basis for a rational exploitation of the BC for targeted delivery.
Blood is the main biological fluid of humans and most animals. Understanding the effects of the biotransformation of nanomaterials in blood plasma on their interactions with cells is fundamental for the evaluation of their health risks and applications. However, there is a lack of related information. The present work found that free radicals and biological molecules in human blood plasma simultaneously drive the formation of a biological corona on biodegraded graphene oxide (GO) nanosheets. Importantly, the above biotransformation affected the interactions of GO with cells. For example, the biotransformed GO induced lower levels of reactive oxygen species and cell ultrastructure damage than did the pristine GO. In addition to the well-known protein corona, the small-molecule corona on the biodegraded GO also plays a critical role in the reduction of GO cytotoxicity through quenching excessive free radical generation. A metabolomics analysis revealed that biotransformation reduced the oxidative stress induced by GO mainly via upregulation of the fatty acid metabolism and downregulation of the galactose metabolism. Overall, the present work clearly shows that the biotransformation of GO in blood plasma influences the nanotoxicity of GO. Compared with pristine nanomaterials, biotransformed materials are more biologically relevant to assessments of their environmental health risks.
We summarized the findings of toxicity studies on graphene-based nanomaterials (GNMs) in laboratory mammals. The inhalation of graphene (GP) and graphene oxide (GO) induced only minimal pulmonary toxicity. Bolus airway exposure to GP and GO caused acute and subchronic pulmonary inflammation. Large-sized GO (L-GO) was more toxic than small-sized GO (S-GO). Intratracheally administered GP passed through the air-blood barrier into the blood and intravenous GO distributed mainly in the lungs, liver, and spleen. S-GO and L-GO mainly accumulated in the liver and lungs, respectively. Limited information showed the potential behavioral, reproductive, and developmental toxicity and genotoxicity of GNMs. There are indications that oxidative stress and inflammation may be involved in the toxicity of GNMs. The surface reactivity, size, and dispersion status of GNMs play an important role in the induction of toxicity and biodistribution of GNMs. Although this review paper provides initial information on the potential toxicity of GNMs, data are still very limited, especially when taking into account the many different types of GNMs and their potential modifications. To fill the data gap, further studies should be performed using laboratory mammals exposed using the route and dose anticipated for human exposure scenarios.
Graphene-based materials have attracted increasing attentions due to their atomically-thick two-dimensional structures, high conductivity, excellent mechanical properties, and large specific surface areas. The combination of biomolecules with graphene-based materials offers a promising way to fabricate novel graphene-biomolecule hybrid nanomaterials with unique functions in biology, medicine, nanotechnology, and materials science. In this review, we focus on a summarization of the recent studies in functionalizing graphene-based materials using different biomolecules, such as DNA, peptide, protein, enzyme, carbohydrate, and virus. The different interactions between graphene and biomolecules at the molecular level are demonstrated and discussed in detail. In addition, the potential applications of the created raphenebiomolecule
nanohybrids in drug delivery, cancer treatment, tissue engineering, biosensor, bioimaging, energy materials,
and other nanotechnological applications are presented. This review will be helpful for knowing the modification of graphene with biomolecules, understanding the interactions between graphene and biomolecules at the molecule level, and designing functional graphene-based nanomaterials for unique properties for various applications.
With mounting evidence that nanomaterials can trigger adverse innate immune responses such as complement activation, there is increasing attention to the development of strategies that mask the complement-activating properties of nanomaterials. The current gold standard to reduce complement activation of nanomaterials is the covalent attachment of polymer coatings on nanomaterial surfaces, even though this strategy provides only moderate protection against complement activation. Akin to protein coronas that form on nanomaterial surfaces in physiological fluids, noncovalent strategies based on protein adsorption would offer a simplified, biomimetic approach to mitigate complement activation. Herein, we demonstrate that precoating graphene-based nanomaterials with purified, natural proteins enables regulatory control of nanomaterial-triggered complement activation. When the graphene-based nanomaterials were coated with complement factor H, nearly complete protection (>90% reduction) against complement activation (a “stealth effect”) was achieved. By contrast, coating the nanomaterials with a passivating layer of bovine or human serum albumins achieved moderate protection (~40% reduction), whereas immunoglobulin G amplified complement activation by several-fold. Taken together, our results demonstrate that surface-bound factor H, as well as serum albumins, can prevent graphene oxide-triggered complement activation, thereby offering a facile approach to inhibit complement activation completely down to naturally occurring levels.
Despite the advances in biomedical applications of nanoparticle (NP) and numerous publications, few NPs have made it to clinical trials and even fewer have reached clinical practice. This wide gap between bench discoveries and clinical applications is mainly because of our limited understanding of the biological identity of NPs. In physiological environments, NPs are coated by a 'protein corona' (PC), critically affecting physiological and therapeutic responses. To date, nearly all studies attempting to characterize the PC have been conducted in vitro. Here, we review recent advances in our understanding of the in vivo PC. We also discuss recent developments of quantitative models to predict biological interactions and how they offer new opportunities for the clinical translation of NPs.
The present work experimentally investigates the interaction of aromatic amino acids viz., tyrosine, tryptophan, and phenylalnine with novel two-dimensional (2D) materials including graphene, graphene oxide (GO), and boron nitride (BN). Photoluminescence, micro-Raman spectroscopy, and cyclic voltammetry were employed to investigate the nature of interactions and possible charge transfer between 2D materials and amino acids. Graphene and GO were found to interact strongly with aromatic amino acids through π−π stacking, charge transfer, and H-bonding. Particularly, it was observed that both physi and chemisorption are prominent in the interactions of GO/graphene with phenylalanine and tryptophan while tyrosine exhibited strong chemisorption on graphene and GO. In contrast, BN exhibited little or no interactions, which could be attributed to localized π-electron clouds around N atoms in BN lattice. Lastly, the adsorption of amino acids on 2D materials was observed to considerably change their biological response in terms of reactive oxygen species generation. More importantly, these changes in the biological response followed the same trends observed in the physi and chemisorption measurements.
In the growing area of nanomedicine, graphene-based materials (GBMs) are some of the most recent explored nanomaterials. For the majority of GBM applications in nanomedicine, the immune system plays a fundamental role. It is necessary to well understand the complexity of the interactions between GBMs, the immune cells and the immune components, and how they could be of advantage for novel effective diagnostic and therapeutic approaches. In this review, we aimed at painting the current picture of GBMs in the background of the immune system. The picture we have drawn looks like a cubist image, a sort of Picasso-like portrait looking at the topic from all perspectives: the challenges (due to the potential toxicity) and the potentiality like the conjugation of GBMs to biomolecules to develop advanced nanomedicine tools. In this context, we have described and discussed: i) the impact of graphene on immune cells; ii) graphene as immunobiosensor; and iii) antibodies conjugated to graphene for tumor targeting.
We investigate the molecular interactions between graphene oxide (GO) and blood plasma proteins, in particular, the influence of GO on the intrinsic fluorescence of these proteins. We observe that GO acts as an efficient quencher of the intrinsic fluorescence of albumin, globulin, and fibrinogen. Interestingly, we also note for the first time that, in addition to the robust fluorescence quenching, GO is capable of selectively amplifying the fluorescence emission of fibrinogen up to approximately 30% or 1.3 fold under certain concentrations but not those of albumin and globulin. We suggest that GO may possibly play a dual role in controlling the intrinsic fluorescence emission of the plasma proteins. Furthermore, this role switching may be influenced by the competition between the aggregation and encapsulation effects. We propose that the GO-induced intrinsic fluorescence quenching is driven by the physical encapsulation of the plasma proteins by GO nanosheets. Contrastingly, the GO-mediated fluorescence amplification is promoted by an aggregation of fibrinogen.
We investigate the molecular interactions between graphene oxide (GO) and human blood plasma proteins. To gain an insight into the bio-physico-chemical activity of GO in biological and biomedical applications, we performed a series of biophysical assays to quantify the molecular interactions between GO with different lateral sizes distribution and the three essential human blood plasma proteins. We elucidate the various aspects of the GO-protein interactions, particularly, the adsorption, binding kinetics and equilibrium, and conformational stability, through the determination of quantitative parameters, such as GO-protein association constants, binding cooperativity, and binding-driven protein structural change. We demonstrate that the molecular interactions between GO and plasma proteins are significantly dependent on the lateral size distribution and mean lateral sizes of the GO nanosheets and their subtle variations may markedly influence the GO-protein interactions. Consequently, we propose the existence of size-dependent molecular interactions between GO nanosheets and plasma proteins, and importantly, the presence of specific critical mean lateral sizes of GO nanosheets in achieving the highest association and fluorescence quenching efficiency of the plasma proteins. We anticipate that this work will offer a sound basis for the design of graphene-based and other related nanomaterials for a plethora of biological and biomedical applications.
Graphene oxide (GO)-based nanocarriers have been frequently studied due to their high drug loading capacity. However, the unsatisfactory biocompatibility of these GO-based nanocarriers hampers their use in clinical settings. This review discusses how each of the physicochemical characteristics (e.g., size, surface area, surface properties, number of layers and particulate states) and surface coatings on GO affect its in vitro and in vivo nanotoxicity. We provide an overview on the effect of GO properties in interactions with cells such as red blood cells, macrophages and cell lines, and experimental organisms including rodents, rabbits and Zebrafish, offering some guidelines for development of safe GO-based nanocarriers. We conclude the paper by outlining the challenges involving GO-based formulations and future perspectives of this research in the biomedical field.
The unique physicochemical properties of two dimensional (2D) graphene oxide (GO) could greatly benefit the biomedical field; however, recent research demonstrated that GO could induce in vitro and in vivo toxicity. We determined the mechanism of GO induced toxicity, and our in vitro experiments revealed that pristine GO could impair cell membrane integrity and functions including regulation of membrane- and cytoskeleton-associated genes, membrane permeability, fluidity and ion channels. Furthermore, GO induced platelet depletion, pro-inflammatory response and pathological changes of lung and liver in mice. To improve the biocompatibility of pristine GO, we prepared a series of GO derivatives including aminated GO (GO-NH2), poly(acrylamide)-functionalized GO (GO-PAM), poly(acrylic acid)-functionalized GO (GO-PAA) and poly(ethylene glycol)-functionalized GO (GO-PEG), and compared their toxicity with pristine GO in vitro and in vivo. Among these GO derivatives, GO-PEG and GO-PAA induced less toxicity than pristine GO, and GO-PAA was the most biocompatible one in vitro and in vivo. The differences in biocompatibility were due to the differential compositions of protein corona, especially IgG, formed on their surfaces that determine their cell membrane interaction and cellular uptake, the extent of platelet depletion in blood, thrombus formation under short-term exposure and the pro-inflammatory effects under long-term exposure. Overall, our combined data delineated the key molecular mechanisms underlying the in vivo and in vitro biological behaviors and toxicity of pristine GO, and identified a safer GO derivative that could be used for future applications.
Benefiting from their unique physicochemical properties, graphene derivatives have attracted great attention in biomedicine. In this study, we carefully engineered graphene oxide (GO) as vaccine adjuvants for immunotherapy using urease B (Ure B) as the model antigen. Ure B is a specific antigen for Helicobacter pylori, which is a class I carcinogen for gastric cancer. Polyethylene glycol (PEG) and various types of polyethylenimine (PEI) were used as coating polymers. Compared with single-polymer modified GOs (GO-PEG and GO-PEI), certain dual-polymer modified GO (GO-PEG-PEI) can act as a positive modulator to promote the maturation of dendritic cells (DCs) and enhance their cytokine secretion through activation of multiple toll-like receptor (TLR) pathways while showing low toxicity. Moreover, this GO-PEG-PEI can serve as an antigen carrier to effectively shuttle antigens into DCs. These two advantages enable GO-PEG-PEI to serve as a novel vaccine adjuvant. In the subsequent in vivo experiments, compared with free Ure B and clinically used aluminum-adjuvant-based vaccine (Alum-Ure B), GO-PEG-PEI-Ure B induces stronger cellular immunity via intradermal administration, suggesting promising applications in cancer immunotherapy. Our work not only presents a novel, highly effective GO-based vaccine nano-adjuvant, but also highlights critical roles of surface chemistry for the rational design of nano-adjuvants.
The non-covalent functionalization of graphene oxide (GO) with chitosan (CS) and dextran (Dex) was successfully developed via layer-by-layer self-assembly technique for anti-cancer drug delivery application. The CS/Dex functionalized GO nanocomposites (GO-CS/Dex) exhibited a diameter of about 300 nm and a thickness of 60 nm. Anti-cancer drug doxorubicin hydrochloride (DOX) was loaded into the nanocomposites via π-π stacking and electrostatic attraction and DOX-loaded nanocomposites exhibited noticeable pH-sensitive DOX release behaviors with release acceleration in acidic environment. The functionalization with CS and Dex not only strongly improved the dispersibility of both GO and DOX loaded GO nanosheets in physiological conditions, but also decreased the non-specific protein adsorption of GO nanosheets, which was beneficial for the biomedical applications. Furthermore, it was observed that the GO-CS/Dex nanocomposites was able to be up-taken by MCF-7 cells and DOX loaded GO-CS/Dex nanocomposites had a strong cytotoxicity to the cancer cells. Overall, GO-CS/Dex could prove to be a suitable candidate for anti-cancer drug delivery.
Current studies have revealed the immune effects of graphene oxide (GO) and have utilized them as vaccine carriers and adjuvants. However, GO easily induces strong oxidative stress and inflammatory reaction at the site of injection. It is very necessary to develop an alternative adjuvant based on graphene oxide derivatives for improving immune responses and decreasing side effects. Carnosine (Car) is an outstanding and safe antioxidant. Herein, the feasibility and efficiency of ultra-small graphene oxide decorated with carnosine as an alternative immune adjuvant were explored. OVA@GO-Car was prepared by simply mixing ovalbumin (OVA, a model antigen) with ultra-small GO covalently modified with carnosine (GO-Car). We investigated the immunological properties of the GO-Car adjuvant in model mice. Results show that OVA@GO-Car can promote robust and durable OVA specific antibody response, increase lymphocyte proliferation efficiency and enhance CD4+ T and CD8+ T cells activation. The presence of Car in GO also probably contribute to enhancing the antigen-specific adaptive immune response through modulating the expression of some cytokines, including IL-6, CXCL1, CCL2 and CSF3. In addition, the safety of GO-Car as an adjuvant was evaluated comprehensively. No symptoms such as allergic response, inflammatory redness swelling, raised surface temperatures, physiological anomalies of blood and remarkable weight changes were observed. Besides, after modification with carnosine, histological damages caused by GO-Car in lung, muscle, kidney and spleen became weaken significantly. This study sufficiently suggest that GO-Car as a safe adjuvant can effectively enhance humoral and innate immune responses against antigens in vivo.
The interaction of graphene oxide (GO) with Bovine Serum Albumin (BSA) in aqueous buffer solution have been investigated with various spectroscopic and imaging techniques. At single molecular resolution this interaction has been performed using Fluorescence Correlation Spectroscopy (FCS) and Fluorescence Lifetime Imaging Microscopy (FLIM) techniques. The conformational dynamics of BSA on GO's influence have been explored by FCS and circular dichroism (CD) spectroscopy. For the FCS studies BSA was labeled covalently by a fluorophore, Alexa Fluor 488. On the addition of GO in phosphate buffer of 10 mM at pH 7.4 the diffusion time (τD) and the hydrodynamic radius (Rh) of BSA increases due to adsorption of BSA. Conformational relaxation time components of native BSA drastically vary with the addition of GO, signifying the change of conformational dynamics of BSA after addition of GO. The adsorption isotherm also indicates significant adsorption of BSA on GO surface. Adsorption of BSA on GO surface has shown in direct images of Atomic Force Microscopy (AFM) and FLIM. Fluorescence quenching study of BSA with addition of GO also indicates that there is strong interaction between BSA and GO.
Nanoparticle (NP) exposure to biological fluids in the body results in protein binding to the NP surface, which forms a protein coating that is called the "protein corona". To simplify studies of protein-NP interactions and protein corona formation, NPs are incubated with biological solutions, such as human serum or human plasma and the effects of this exposure are characterized in vitro. Yet, how NP exposure to these two different biological milieus affects protein corona composition and cell response has not been investigated. Here, we explore the differences between the protein coronas that form when NPs are incubated in human serum versus human plasma. NP characterization indicated that NPs that were exposed to human plasma had higher amounts of proteins bound to their surfaces, and were slightly larger in size than those exposed to human serum. In addition, significant differences in corona composition were also detected with gel electrophoresis and liquid chromatography-mass spectrometry/mass spectrometry, where a higher fraction of coagulation proteins and complement factors were found on the plasma-exposed NPs. Flow cytometry and confocal microscopy showed that the uptake of plasma-exposed NPs was higher than that of serum-exposed NPs by RAW 264.7 macrophage immune cells, but not by NIH 3T3 fibroblast cells. This difference is likely due to the elevated amounts of opsinins, such as fibrinogen, on the surfaces of the NPs exposed to plasma, but not serum, because these components trigger NP internalization by immune cells. Because the human plasma better mimics the composition of the in vivo environment, namely blood, in vitro protein corona studies should employ human plasma, and not human serum, so the biological phenomena that is observed is more similar to that occurring in vivo.
