Systematic Study of Various Functionalization Steps for Ultrasensitive Detection of SARS-CoV-2 with Direct Laser-Functionalized Au-LIG Electrochemical Sensors.

The 2019 coronavirus (COVID-19) pandemic impaired global health, disrupted society, and slowed the economy. Early detection of the infection using highly sensitive diagnostics is crucial in preventing the disease's spread. In this paper, we demonstrate electrochemical sensors based on laser induced graphene (LIG) functionalized directly with gold (Au) nanostructures for the detection of SARS-CoV-2 with an outstanding limit of detection (LOD) of ∼1.2 ag·mL-1. To achieve the optimum performance, we explored various functionalization parameters to elucidate their impact on the LOD, sensitivity, and linearity. Specifically, we investigated the effect of (i) gold precursor concentration, (ii) cross-linker chemistry, (iii) cross-linker and antibody incubation conditions, and (iv) antigen-sensor interaction (diffusion-dominated incubation vs pipette-mixing), as there is a lack of a systematic study of these parameters. Our benchmarking analysis highlights the critical role of the antigen-sensor interaction and cross-linker chemistry. We showed that pipette-mixing enhances sensitivity and LOD by more than 1.6- and 5.5-fold, respectively, and also enables multimodal readout compared to diffusion-dominated incubation. Moreover, the PBA/Sulfo-NHS: EDC cross-linker improves the sensitivity and LOD compared to PBASE. The sensors demonstrate excellent selectivity against other viruses, including HCoV-229E, HCoV-OC43, HCoV-NL63, and influenza H5N1. Beyond the ability to detect antigen fragments, our sensors enable the detection of antigen-coated virion mimics (which are a better representative of the real infection) down to an ultralow concentration of ∼5 particles·mL-1.

Recent advances in graphene-based electroanalytical devices for healthcare applications.

Graphene, with its outstanding mechanical, electrical, and biocompatible properties, stands out as an emerging nanomaterial for healthcare applications, especially in building electroanalytical biodevices. With the rising prevalence of chronic diseases and infectious diseases, such as the COVID-19 pandemic, the demand for point-of-care testing and remote patient monitoring has never been greater. Owing to their portability, ease of manufacturing, scalability, and rapid and sensitive response, electroanalytical devices excel in these settings for improved healthcare accessibility, especially in resource-limited settings. The development of different synthesis methods yielding large-scale graphene and its derivatives with controllable properties, compatible with device manufacturing - from lithography to various printing methods - and tunable electrical, chemical, and electrochemical properties make it an attractive candidate for electroanalytical devices. This review article sheds light on how graphene-based devices can be transformative in addressing pressing healthcare needs, ranging from the fundamental understanding of biology in in vivo and ex vivo studies to early disease detection and management using in vitro assays and wearable devices. In particular, the article provides a special focus on (i) synthesis and functionalization techniques, emphasizing their suitability for scalable integration into devices, (ii) various transduction methods to design diverse electroanalytical device architectures, (iii) a myriad of applications using devices based on graphene, its derivatives, and hybrids with other nanomaterials, and (iv) emerging technologies at the intersection of device engineering and advanced data analytics. Finally, some of the major hurdles that graphene biodevices face for translation into clinical applications are discussed.

Linker-free Functionalization of Phosphorene Nanosheets by Sialic Acid Biomolecules.

The importance of sialic acid on cell functions has been recently unveiled, and consequently, great attention has been paid to its interaction with tumor cells. In this line of research, we have realized phosphorene nanosheets functionalized with sialic acid molecules for biological applications with no need for another linker molecule. The formation of phosphorene sheets is feasible by using hydrogen plasma treatment and conversion of amorphous phosphorus on silicon substrates into highly crystalline nanosheets. Through immersion of these freshly prepared nanosheets into an aqueous solution containing sialic acid molecules, the formation of chemical binding between biomolecules and P atoms is initiated to form a carpet-like coverage. We have studied these structures by using Raman spectroscopy, electron microscopy, FTIR-ATR spectroscopy, and X-ray photoelectron spectroscopy. While XPS supports the passivation of sialic-activated phosphorene nanosheets (SAP) against oxidation in air or aqueous solutions, the FTIR analysis corroborates the evolution of P-O-C and P-C bonds between such biomolecules and the sheet surface. Moreover, the high-resolution TEM images demonstrate a considerable reduction in the lattice spacing from 0.32 nm for pristine phosphorene to 0.30 nm. Similarly, Raman spectroscopy depicts a shift in A2g in-plane vibrations, owing to the evolution of stress in the passivated sheets. To investigate their biocompatibility, we examined the toxicity of these bioactivated structures and observed no or little sign of toxicity. For the latter evaluation, we exploited MTT, flow cytometry, and animal models for in vivo investigations.

Electrochemical Sensors Based on MoSx -Functionalized Laser-Induced Graphene for Real-Time Monitoring of Phenazines Produced by Pseudomonas aeruginosa.

Pseudomonas aeruginosa (P. aeruginosa) is an opportunistic pathogen causing infections in blood and implanted devices. Traditional identification methods take more than 24 h to produce results. Molecular biology methods expedite detection, but require an advanced skill set. To address these challenges, this work demonstrates functionalization of laser-induced graphene (LIG) for developing flexible electrochemical sensors for P. aeruginosa based on phenazines. Electrodeposition as a facile approach is used to functionalize LIG with molybdenum polysulfide (MoSx ). The sensor's limit of detection (LOD), sensitivity, and specificity are determined in broth, agar, and wound simulating medium (WSM). Control experiments with Escherichia coli, which does not produce phenazines, demonstrate specificity of sensors for P. aeruginosa. The LOD for pyocyanin (PYO) and phenazine-1-carboxylic acid (PCA) is 0.19 × 10-6  and 1.2 × 10-6  m, respectively. Furthermore, the highly stable sensors enable real-time monitoring of P. aeruginosa biofilms over several days. Comparing square wave voltammetry data over time shows time-dependent generation of phenazines. In particular, two configurations-"Normal" and "Flipped"-are studied, showing that the phenazines time dynamics vary depending on how cells interact with sensors. The reported results demonstrate the potential of the developed sensors for integration with wound dressings for early diagnosis of P. aeruginosa infection.

Electrochemical generation of microbubbles by carbon nanotube interdigital electrodes to increase the permeability and material uptakes of cancer cells.

Artificial cavitation as a prerequisite of sonoporation, plays an important role on the ultrasound (US) assisted drug delivery systems. In this study, we have proposed a new method of microbubble (MB) generation by local electrolysis of the medium. An integrated interdigital array of three-electrode system was designed and patterned on a nickel-coated quartz substrate and then, a short DC electrical pulse was applied that consequently resulted in distributed generation of microbubbles at the periphery of the electrodes. Growth of the carbon nanotube (CNT) nanostructures on the surface of the electrodes approximately increased the number of generated microbubbles up to 7-fold and decreased their average size from ∼20 µm for bare to ∼7 µm for CNT electrodes. After optimizing the three-electrode system, biocompatibility assays of the CNT electrodes stimulated by DC electrical micropulses were conducted. Finally, the effect of the proposed method on the sonoporation efficiency and drug uptake of breast cells were assessed using cell cycle and Annexin V/PI flow cytometry analysis. These results show the potential of electrochemical generation of MBs by CNT electrodes as an easy, available and promising technique for artificial cavitation and ultrasound assisted drug delivery.