A lipid membrane supported on an artificial extracellular matrix made of polyelectrolyte multilayers: towards nanoarchitectonics at the cellular interface.

To implement a specific function, cells recognize multiple physical and chemical cues and exhibit molecular responses at their interfaces - the boundary regions between the cell lipid-based membrane and the surrounding extracellular matrix (ECM). Mimicking the cellular external microenvironment presents a big challenge in nanoarchitectonics due to the complexity of the ECM and lipid membrane fragility. This study reports an approach for the assembly of a lipid bilayer, mimicking the cellular membrane, placed on top of a polyelectrolyte multilayer cushion made of hyaluronic acid and poly-L-lysine - a nanostructured biomaterial, which represents a 3D artificial ECM. Model proteins, lysozyme and α-lactalbumin, (which have similar molecular masses but carry opposite net charges) have been employed as soluble signalling molecules to probe their interaction with these hybrids. The formation of a lipid bilayer and the intermolecular interactions in the hybrid structure are monitored using a quartz crystal microbalance and confocal fluorescence microscopy. Electrostatic interactions between poly-L-lysine and the externally added proteins govern the transport of proteins into the hybrid. Designed ECM-cell mimicking hybrids open up new avenues for modelling a broad range of cell membranes and ECM and their associated phenomena, which can be used as a tool for synthetic biology and drug screening.

Self-degrading graphene sheets for tumor therapy.

Low biodegradability of graphene derivatives and related health risks are the main limiting factors for their in vivo biomedical applications. Here, we present the synthesis of enzyme-functionalized graphene sheets with self-degrading properties under physiological conditions and their applications in tumor therapy. The synergistic enzyme cascade glucose oxidase and myeloperoxidase are covalently conjugated to the surface of graphene sheets and two-dimensional (2D) platforms are obtained that can produce sodium hypochlorite from glucose. The enzyme-functionalized graphene sheets with up to 289 nm average size are degraded into small pieces (≤40 nm) by incubation under physiological conditions for 24 h. Biodegradable graphene sheets are further loaded with doxorubicin and their ability for tumor therapy is evaluated in vitro and in vivo. The laser-triggered release of doxorubicin in combination with the enzymatic activity of the functionalized graphene sheets results in a synergistic antitumor activity. Taking advantage of their neutrophil-like activity, fast biodegradability, high photo- and chemotherapeutic effects, the novel two-dimensional nanoplatforms can be used for tumor therapeutic applications.

Functionalized nanographene sheets with high antiviral activity through synergistic electrostatic and hydrophobic interactions.

As resistance to traditional drugs emerges for treatment of virus infections, the need for new methods for virus inhibition increases. Graphene derivatives with large surface areas have shown strong activity against different viruses. However, the inability of current synthetic protocols to accurately manipulate the structure of graphene sheets in order to control their antiviral activity remains a major challenge. In this work, a series of graphene derivatives with defined polyglycerol sulfate and fatty amine functionalities have been synthesized and their interactions with herpes simplex virus type 1 (HSV-1) are investigated. While electrostatic interactions between polyglycerol sulfate and virus particles trigger the binding of graphene to virus, alkyl chains induce a high antiviral activity by secondary hydrophobic interactions. Among graphene sheets with a broad range of alkyl chains, (C3-C18), the C12-functionalized sheets showed the highest antiviral activity, indicating the optimum synergistic effect between electrostatic and hydrophobic interactions, but this derivative was toxic against the Vero cell line. In contrast, sheets functionalized with C6- and C9-alkyl chains showed low toxicity against Vero cells and a synergistic inhibition of HSV-1. This study shows that antiviral agents against HSV-1 can be obtained by controlled and stepwise functionalization of graphene sheets and may be developed into antiviral agents for future biomedical applications.

Scalable Production of Nanographene and Doping via Nondestructive Covalent Functionalization.

A new method for top-down, one-pot, gram-scale production of high quality nanographene by incubating graphite in a dilute sodium hypochlorite solution at only 40 °C is reported here. The produced sheets have only 4 at% oxygen content, comparable with nanographene grown by chemical vapor deposition. The nanographene sheets are covalently functionalized using a nondestructive nitrene [2+1] cycloaddition reaction that preserves their π-conjugated system. Statistical analyses of Raman spectroscopy and X-ray photoelectron spectroscopy indicate a low number of sp3 carbon atoms on the order of 2% before and 4% after covalent functionalization. The nanographene sheets are significantly more conductive than conventionally prepared nanographene oxide, and conductivity further increases after covalent functionalization. The observed doping effects and theoretical studies suggest sp2 hybridization for the carbon atoms involved in the [2+1] cycloaddition reaction leading to preservation of the π-conjugated system and enhancing conductivity via n-type doping through the bridging N-atom. These methods are easily scalable, which opens the door to a mild and efficient process to produce high quality nanographenes and covalently functionalize them while retaining or improving their physicochemical properties.

Multivalent Interactions between 2D Nanomaterials and Biointerfaces.

2D nanomaterials, particularly graphene, offer many fascinating physicochemical properties that have generated exciting visions of future biological applications. In order to capitalize on the potential of 2D nanomaterials in this field, a full understanding of their interactions with biointerfaces is crucial. The uptake pathways, toxicity, long-term fate of 2D nanomaterials in biological systems, and their interactions with the living systems are fundamental questions that must be understood. Here, the latest progress is summarized, with a focus on pathogen, mammalian cell, and tissue interactions. The cellular uptake pathways of graphene derivatives will be discussed, along with health risks, and interactions with membranes-including bacteria and viruses-and the role of chemical structure and modifications. Other novel 2D nanomaterials with potential biomedical applications, such as transition-metal dichalcogenides, transition-metal oxide, and black phosphorus will be discussed at the end of this review.

Directed Graphene-Based Nanoplatforms for Hyperthermia: Overcoming Multiple Drug Resistance.

Multidrug resistance (MDR), which leads tumors resistance to traditional anticancer drugs, can cause the failure of chemotherapy treatments. Herein, we present a new way to overcome this problem using smart multifunctional graphene-based drug delivery systems which can target subcellular organelles and show synergistic hyperthermia and chemotherapy. Mitochondria-targeting ligands are conjugated onto the doxorubicin-loaded, polyglycerol-covered nanographene sheets to actively accumulate them inside the mitochondria after charge-mediated cellular internalization. Upon near-infrared (NIR) irradiation, adenosine triphosphate (ATP) synthesis and mitochondrial function were inhibited and doxorubicin released into the cellular interior. The hyperthermia-accelerated drug release led to a highly selective anticancer efficiency, confirmed by in vitro and in vivo experiments.