Graphene/Polymer Nanocomposites for Human Health Monitoring
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This entry is adapted from the peer-reviewed paper 10.3390/polym14051030

Graphene/polymer nanocomposites (GPNs) are largely explored in the development of sensing devices to monitor human health parameters, due to the excellent electrical and mechanical properties of the graphene filler combined with the chemical versatility of the surrounding polymer matrix.

graphene polymers sensors nanocomposites human health monitoring

1. Graphene/Polymer Nanocomposites: Fabrication and Properties

Graphene/polymer nanocomposites (GPNs) with polymer matrix are commonly fabricated following three different methods: solution blending, in situ polymerization and melt mixing [1][2]. The most largely used technique is solution blending, which involves the solubilization of the polymer in a suitable solvent and the mixing with graphene to form a homogeneous dispersion. Generally, polymers such as polystyrene, polycarbonate, polyacrylamide, polyimides and poly(methyl methacrylate) are mixed with graphene oxide [3][4][5], which can be previously functionalized with isocyanates, alkylamine or alkyl-chlorosilanes in order to improve its dispersibility in organic solvents.

The fabrication of GPNs by in situ polymerization is based on the polymerization of the matrix in the presence of the selected filler, starting from a mixture of monomer and reinforcement [1]. Typically, this approach allows to obtain a good grade of dispersion of graphene-based nanofillers avoiding their previous exfoliation. In the melt mixing technique, the filler is dispersed in the polymer matrix exploiting high temperatures and shear forces [6]. The polymer phase is brought to melting at high temperatures, thus facilitating the dispersion or intercalation of the graphene oxide nanoplatelets without the use of organic, often toxic, solvents.

The properties of the GPN nanocomposites are strictly related to the spatial distribution and alignment of the graphene nanofiller, and to its interfacial adhesion with the polymer phase. In particular, GPNs with low loadings of functionalized graphene sheets generally exhibit a shift in the glass transition temperature [4], if compared with the value of the uncharged polymer. This behavior can be ascribed to a reduced mobility of the polymer chains at the interface between the filler and the matrix [7][8]. Therefore, the effect of constraint applied on the chains can directly induce an increase in glass transition temperature [9].

In terms of thermal conductivity, the performance of GPNs can be evaluated referring to the 2D geometry of the graphene fillers. These are characterized by a lower interfacial thermal resistance that provides a higher thermal conductivity to the host polymer matrix [10][11]. Nevertheless, the 2D structure can be source of anisotropy in the nanocomposites arrangement, for which the in-plane thermal conductivity results as much as ten times higher than the cross-plane conductivity [12]. This is typically evaluated following the percolation theory, therefore considering phonons as the main mode for thermal conduction in polymers. Covalent bonding between the filler and the polymer matrix can reduce phonon scattering at the interface leading to an overall enhancement of the GPN thermal conductivity [13].

The electrical conductivity behavior of GPNs can be analyzed considering the influence of different factors and their overall effect. In particular, the characteristics of the specific graphene-based filler, such as its aspect ratio and morphology, as well as its inter-sheet junction, can affect the electrical performances of GPNs [14]. In the same way, processing and dispersion, and the related state of aggregation and alignment of the nanoparticles concur to determine the electrical behavior of the resulting nanocomposites [15]. Several theoretical models and experiments were to assess the role of nanofiller shape, geometry and state of dispersion on the percolation threshold of graphene/polymer nanocomposites [16][17].

As mentioned above, the overall performance of polymer-based nanocomposite materials can be related to the quality and stability of the polymer/nanofiller interphase region. Typically, the physical and mechanical properties and the chemical composition of this region are different from those of the bulk polymer matrix [18][19]. In the case of an interphase stiffer than the surrounding polymer, this can result in higher overall stiffness and strength of the composite, but with lower resistance to fracture [20]. The interphase properties can affect the mechanical behavior of the nanocomposites also depending on the morphology and size of this region. In fact, several studies show that its thickness can be tailored with the aim to achieve both higher strength and improved toughness of the resulting nanocomposites [21][22][23]. Force-modulation atomic force microscopy (AFM) and nanoindentation are common techniques that are used to investigate the interphase and its properties [18][24]. In particular, AFM phase imaging is currently considered a useful tool to evaluate the thickness and the relative stiffness of the interphase, since it involves much lower interaction forces between the probe and the sample than force modulation or nanoindentation [20]. The arrangement of graphene-based fillers inside the polymer matrix is also investigated in order to assess the state of dispersion at the microstructural level and its impact on the nanocomposite properties. Results reveal that graphene-based fillers, such as graphene oxide or graphene nanoplatelets, can arrange differently in the host polymer, originating structural states that can be classified as stacked, intercalated or exfoliated [1].

The intercalated state can be considered a particular stacked structure, which is characterized by a greater interlayer spacing, but within a few nanometers [25]. Generally, in the exfoliated structure, graphene nanoplatelets have the largest interfacial contact with the polymer matrix, and this allows to improve the performances of the composites in different ways. Due to the interactions with the matrix, the exfoliated phase can exhibit a curved shape. The rumpled shape assumed by the filler can result in a mechanical interlocking acting as a possible mechanism of strengthening. The compatibility between the host polymer and the nanoplatelets is one of the major factors determining the filler morphology in the matrix: the nanoplatelets are characterized by a more extended conformation for high polymer/filler affinity or, conversely, a crumpled conformation when the affinity decreases [26]. Finally, the processing method used to fabricate the nanocomposite also affects its microstructure to a great extent: solution mixing or in situ polymerization generally induce an exfoliated and randomly oriented status of the nanoplatelets, whereas from the melt mixing technique a more oriented and intercalated or stacked structure of the nanoplatelets is generated [27].

2. Graphene/Polymer Nanocomposites: Applications in Human Health Monitoring

Many sensors based on nanocomposite materials find applications in the monitoring of human health parameters [28][29]. In this perspective, the sensing devices need to be comfortable to wear, biocompatible, and lightweight [30][31][32]. They need to interface with human body, showing at the same time high selectivity and sensitivity to detect and quantify specific signals or analytes.

Graphene, graphene oxide and chemically modified graphene are widely employed to fabricate nanocomposites suitable for detecting biological analytes, such as uric acid and ascorbic acid [33], hydroquinone and catechol [34], and nucleic bases [35][36]. The presence of functional groups on the nanocomposite surface is fundamental to create hydrogen bonds with the analytes, so the strength of these bonds and the distance between the interaction sites and the reaction center make possible the discrimination of the analytes.

DNA molecules can be immobilized on graphene surface by physical adsorption or chemical binding, thus creating sensitive platforms where each binding event with the analyte can be detected through the changes of the electric or electrochemical properties of these platforms [37][38]. Noncovalent interactions can be promoted through the physical adsorption, involving π-π stacking interactions between the DNA nucleobases and the aromatic surface of graphene. In particular, in the case of single-stranded DNA (ssDNA), stable aqueous dispersions of graphene/DNA can be obtained, without traces of sedimentation for months [39]. Double-stranded DNA (dsDNA) is also used as dispersing agent for graphene nanoplatelets. However, less stable aqueous solutions are obtained due to the weaker hydrophobic interactions arising from the base pairing of the nucleobases. Nevertheless, the graphene/dsDNA affinity can be significantly enhanced by further functionalizing graphene oxide with polar groups, which are able to establish electrostatic interactions with the DNA bases. The immobilization of DNA on graphene through covalent bonds is generally carried out after functionalizing the DNA with amino groups, which are able to interact with the graphene oxide surface via carbodiimide chemistry [40]. In particular, amine-terminated ssDNA can be linked to the surface of graphene oxide directly or through the involvement of specific molecules that act as carriers.

Single-stranded DNA was covalently immobilized on a polyaniline/graphene (PAN/GN) nanocomposite, which was applied onto a glassy carbon electrode (GCE) and used for HIV-1 gene detection [41]. In particular, the negatively-charged phosphate backbone of the HIV-1 binds to the sensitive surface via π-π∗ stacking interactions. The hybridization between the ssDNA probe and the target HIV-1 generates double-stranded DNA (dsDNA), which induces an increase of the electron transfer resistance that can be correlated with the concentration of the gene. The sensitivity and the selectivity of this nanocomposite were tested, and a low detection limit of 1.0 × 10−16 M for the target HIV-1 was measured. 

The development of new composite materials using biomolecules, such as enzymes, has allowed to extend even more the field of sensing applications in medical diagnosis and bio-industrial analysis. Several sensitive nanocomposites have been realized using natural polymers as matrix, such as gelatin, alginate and chitosan, due to their intrinsic biodegradability and biocompatibility that make it suitable for biomedical applications [42]. In particular, chitosan was combined with graphene to develop nanocomposite materials with sensing properties useful for monitoring human health [43][44][45].

Xie et al. developed an immunosensor based on graphene and chitosan-modified screen-printed carbon electrode (SPCE) [46]. The phospho-p53 capture antibody was adsorbed on the surface of the graphene-chitosan/SPCE. A sandwich immunocomplex was formed between the targeted phospho-p5315 antigen, the phospho-p53 capture antibody, the antigen, and biotinylated phospho-p5315 detection antibody, which was previously marked with horseradish peroxidase (HRP). The high surface area of graphene allowed to immobilize a large amount of capture antibody, increasing the sensitivity of this nanocomposite immunosensor.

Nanocomposite films based on glucose oxidase (GOD), platinum (Pt), functional graphene sheets (FGS) and chitosan were developed for glucose sensing [47]. The electrocatalytic action of FGS and Pt nanoparticles towards hydrogen peroxide (H2O2) was exploited to obtain a sensitive biosensor with a detection limit of 0.6 μM of glucose. The performance of this type of sensor can be ascribed to the large surface area and the fast electron transfer of graphene and Pt nanoparticles. This sensor showed good reproducibility and long-term stability, with a negligible response to other compounds such as ascorbic acid and uric acid. The GOD/Pt/FGS/chitosan sensitive nanocomposite can be useful for both clinical and home-care devices for a rapid monitoring of glucose.

Glucose sensing was also performed with composite films made of graphene, chitosan and uric acid, which were deposited onto glassy carbon electrodes [44]. A molecularly imprinted electrochemical sensor was obtained, and its sensitivity mechanism was analyzed by electrochemical impedance spectroscopy and chronocoulometric methods. A comparison between graphene-doped and undoped sensors was carried out, with results demonstrating an improvement in terms of sensitivity due to the high surface area and good electronic conduction of graphene.

Sensitive films based on EDTA-modified reduced graphene (EDTA-RG) and Nafion were fabricated and tested as dopamine detectors [48]. Graphene was chemically modified by silanization using N-(trimethoxysilylpropyl) ethylenediamine triacetic acid (EDTA-silane). The selectivity was investigated by using dopamine and ascorbic acid. Experimental tests demonstrated that the sulfuric groups of Nafion and the carboxylic groups of EDTA-RG interfere with the diffusion of ascorbic acid, thus enabling the selective detection of dopamine.

More recently, biosensing has seen advances towards more complex structures that are able to enhance the overall sensitivity of the detecting surface. A sensing composite material was realized using fractal nanoplatinum with a cauliflower-like morphology, which was developed on a reduced graphene oxide paper [49]. Platinum was electrodeposited on the graphene-nanocellulose sheets using pulsed sonoelectrodeposition. As a result, a conductive nanocomposite paper with a high electroactive surface was obtained and then functionalized using glucose oxidase (via chitosan encapsulation) or RNA aptamer (via covalent linking). In this way, the material sensitivity towards glucose or Escherichia coli bacteria can be activated. Depending on the type of the enzyme selected, good performances in terms of sensitivity and response times were obtained.

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