Adaptive in vivo device for theranostics of inflammation: Real-time monitoring of interferon-γ and aspirin.

Cytokines mediate and control immune and inflammatory responses. Complex interactions exist among cytokines, inflammation, and the innate and adaptive immune responses in maintaining homeostasis, health, and well-being. On-demand, local delivery of anti-inflammatory drugs to target tissues provides an approach for more effective drug dosing while reducing the adverse effects of systemic drug delivery. This work demonstrates a proof-of-concept theranostic approach for inflammation based on analyte-kissing induced signaling, whereby a drug (in this report, aspirin) can be released upon the detection of a target level of a proinflammatory cytokine (i.e., interferon-γ (IFN-γ)) in real time. The structure-switching aptamer-based biosensor described here is capable of quantitatively and dynamically detecting IFN-γ both in vitro and in vivo with a sensitivity of 10 pg mL-1. Moreover, the released aspirin triggered by the immunoregulatory cytokine IFN-γ is able to inhibit inflammation in a rat model, and the release of aspirin can be quantitatively controlled. The data reported here provide a new and promising strategy for the in vivo detection of proinflammatory cytokines and the subsequent therapeutic delivery of anti-inflammatory molecules. This universal theranostic platform is expected to have great potential for patient-specific personalized medicine. STATEMENT OF SIGNIFICANCE: We developed an adaptive in vivo sensing device whereby a drug, aspirin, can be released upon the detection of a proinflammatory cytokine, interferon-γ (IFN-γ), in real time with a sensitivity of 10 pg mL-1. Moreover, the aspirin triggered by IFN-γ depressed inflammation in the rat model and was delivered indirectly through blood and cerebrospinal fluid or directly to the inflammation tissue or organ without adverse gastrointestinal effects observed in the liver and kidney. We envision that, for the first time, patients with chronic inflammatory disease can receive the right intervention and treatment at the right time. Additionally, this technology may empower patients to monitor their personalized health and disease management program, allowing real-time diagnostics, disease monitoring, and precise and effective treatments.

On-chip structure-switching aptamer-modified magnetic nanobeads for the continuous monitoring of interferon-gamma ex vivo.

Cytokines are cell signaling molecules that indicate the health status of the body. In this study, we developed a microfluidic device integrated with structure-switching aptamers capable of continuously tracking the concentration of the cytokine interferon gamma (IFN-γ) in cell culture medium and blood serum. First, a ferrocene (Fc)-labeled structure-switching signaling aptamer with a hairpin structure targeting IFN-γ was immobilized on magnetic nanobeads by the strongest noncovalent interactions between streptavidin and biotin. The aptamer-modified magnetic nanobeads were trapped on a customized microfluidic chip by a magnetic field to form the sensing interface. The binding of IFN-γ could trigger the hairpin structure of the aptamer to unfold, pushing Fc redox molecules away from the sensing interface and consequently switching off the electrochemical signal. The change in the redox current of Fc was quantitatively related to the concentration of IFN-γ in a linear range of 10-500 pg mL-1 and with the lowest detection limit of 6 pg mL-1. This microfluidic device was specific to IFN-γ in the presence of overabundant serum proteins and allowed the continuous monitoring of IFN-γ without adding exogenous reagents. It provided a universal point-of-care biosensing platform for the real-time detection of a spectrum of analytes.

Graphene Oxide Based Recyclable in Vivo Device for Amperometric Monitoring of Interferon-γ in Inflammatory Mice.

Cytokine sensing is challenging due to their typically low abundances in physiological conditions. Nanomaterial fabricated interfaces demonstrated unique advantages in ultrasensitive sensing. Here, we demonstrate an amperometric sensing device based on graphene oxide (GO) and structure-switching aptamers for long-term detection of cytokines in a living organism. The device incorporates a single layer of GO acting as a signal amplifier on glassy carbon electrodes. The hairpin aptamers specific to interferon-γ (IFN-γ), which were loaded with redox probes, are covalently attached to GO to serve as biorecognition moieties. IFN-γ was able to trigger the configuration change of aptamers while releasing the trapped redox probes to introduce the electrochemical signal. This in vivo device was capable of quantitatively and dynamically detecting IFN-γ down to 1.3 pg mL-1 secreted by immune cells in cell culture medium with no baseline drift even at a high concentration of other nonspecific proteins. The biocompatible devices were also implanted into subcutaneous tissue of enteritis mice, where they performed precise detection of IFN-γ over 48 h without using physical barriers or active drift correction algorithms. Moreover, the device could be reused even after multiple rounds of regeneration of the sensing interface.

Graphene Oxide Signal Reporter Based Multifunctional Immunosensing Platform for Amperometric Profiling of Multiple Cytokines in Serum.

Cytokines are small proteins and form complicated cytokine networks to report the status of our health. Thus, accurate profiling and sensitive quantification of multiple cytokines is essential to have a comprehensive and accurate understanding of the complex physiological and pathological conditions in the body. In this study, we demonstrated a robust electrochemical immunosensor for the simultaneous detection of three cytokines IL-6, IL-1ß, and TNF-α. First, graphene oxides (GO) were loaded with redox probes nile blue (NB), methyl blue (MB), and ferrocene (Fc), followed by covalent attachment of anti-cytokine antibodies for IL-6, IL-1ß, and TNF-α, respectively, to obtain Ab2-GO-NB, Ab2-GO-MB, and Ab2-GO-Fc, acting as the signal reporters. The sensing interface was fabricated by attachment of mixed layers of 4-carboxylic phenyl and 4-aminophenyl phosphorylcholine (PPC) to glassy carbon surfaces. After that, the capture monoclonal antibody for IL-6, IL-1ß, and TNF-α was modified to the carboxylic acid terminated sensing interface. And finally a sandwich assay was developed. The quantitative detection of three cytokines was achieved by observing the change in electrochemical signal from signal reporters Ab2-GO-NB, Ab2-GO-MB, and Ab2-GO-Fc. The designed system has been successfully used for detection of three cytokines (IL-6, IL-1ß, and TNF-α) simultaneously with desirable performance in sensitivity, selectivity, and stability, and recovery of 93.6%-105.5% was achieved for determining cytokines spiked in the whole mouse serum.

Graphene Oxide Thin Film with Dual Function Integrated into a Nanosandwich Device for in Vivo Monitoring of Interleukin-6.

Graphene oxide (GO), with its exceptional physical and chemical properties and biocompatibility, holds a tremendous potential for sensing applications. In this study, GO, acting both as the electron-transfer bridge and the signal reporter, was attached on the interface to develop a label-free electrochemical nanosandwich device for detection of interleukin-6 (IL-6). First, a single layer of GO was covalently modified on gold electrodes, followed by attachment of anti-IL-6 capture antibody to form the sensing interface. The 4-aminophenyl phosphorylcholine was further attached to the surface of GO to minimize nonspecific protein adsorption. For reporting the presence of analyte, the anti-IL-6 detection antibody was covalently modified to the GO, which has been integrated with the redox probe Nile blue (NB). Finally, a nanosandwich assay was fabricated on gold surfaces for detection of IL-6 on the basis of the electrochemical signal of NB. The prepared nanosandwiches demonstrated high selectivity and stability for detection of IL-6 over the range of 1-300 pg mL-1 with the lowest detectable concentration of 1 pg mL-1. The device was successfully used for monitoring of IL-6 secretion in RAW cells and live mice. By tailoring the GO surface with functional components, such devices were able to detect the analyte in vivo without causing inflammatory response.

Advances on Aryldiazonium Salt Chemistry Based Interfacial Fabrication for Sensing Applications.

Aryldiazonium salts as coupling agents for surface chemistry have evidenced their wide applications for the development of sensors. Combined with advances in nanomaterials, current trends in sensor science and a variety of particular advantages of aryldiazonium salt chemistry in sensing have driven the aryldiazonium salt-based sensing strategies to grow at an astonishing pace. This review focuses on the advances in the use of aryldiazonium salts for modifying interfaces in sensors and biosensors during the past decade. It will first summarize the current methods for modification of interfaces with aryldiazonium salts, and then discuss the sensing applications of aryldiazonium salts modified on different transducers (bulky solid electrodes, nanomaterials modified bulky solid electrodes, and nanoparticles). Finally, the challenges and perspectives that aryldiazonium salt chemistry is facing in sensing applications are critically discussed.

Decoration of Reduced Graphene Oxide Nanosheets with Aryldiazonium Salts and Gold Nanoparticles toward a Label-Free Amperometric Immunosensor for Detecting Cytokine Tumor Necrosis Factor-α in Live Cells.

In this study, a label-free electrochemical immunosensor was developed for detection of cytokine tumor necrosis factor-alpha (TNF-α). First, AuNPs loaded reduced graphene oxides nanocomposites (RGO-ph-AuNP) were prepared, and then, a mixed layer of 4-carbxyphenyl and 4-aminophenyl phosphorylcholine (PPC) was modified to the surface of AuNPs for the subsequent modification of anti-TNF-α capture antibody (Ab1) to form the capture surface (Au-RGO-ph-AuNP-ph-PPC(-ph-COOH)) for the analyte TNF-α with the antifouling property. For reporting the presence of analyte, the anti-TNF-α detection antibody (Ab2) was modified to the graphene oxides which have been modified with the 4-ferrocenylaniline through diazonium chemistry to form Ab2-GO-ph-Fc. Then, a sandwich assay was formed on gold surfaces for the quantitative detection of TNF-α based on the electrochemical signal of ferrocene. X-ray photoelectron spectra (XPS), transmission electron microscopy (TEM), Fourier transform infrared spectroscopy (FT-IR), UV-vis, and electrochemistry were used for characterization of the stepwise fabrications on the interface. The prepared electrochemical immunosensor was successfully used for the detection of TNF-α over the range of 0.1-150 pg mL-1. The lowest detection limit of this immunosensor is 0.1 pg mL-1 TNF-α in 50 mM phosphate buffer at pH 7.0. The fabricated immunosensor provided high selectivity and stability and can be used to detect TNF-α secreted by live BV-2 cells with comparable accuracy to enzyme-linked immunosorbent assay (ELISA) but with lower limit of detection.

Nanocomposites of gold nanoparticles and graphene oxide towards an stable label-free electrochemical immunosensor for detection of cardiac marker troponin-I.

A stable label-free amperometric immunosensor is presented based on gold nanoparticles and graphene oxide nanocomposites for detection of cardiac troponin-I in the early diagnosis of myocardial infarction. For designing of the sensing platform, firstly the nanocomposites based on GO and AuNPs were prepared and anchored on electrode surfaces. The formed nanocomposites provided a platform with big surface area for loading anti-cTnI capture antibody, and worked as a bridge for fast electron transfer subsequently increased the sensitivity. Moreover, the linkages between AuNP, GO, and electrodes were based on covalent bonding by aryldiazonium salt coupling chemistry, which favors the stability of the sensing interface. Finally, the anti-cTnI detection antibody was immobilized on GO tailored with ferrocene molecules, functioning as the signal reporter for the detection of cTnI. The modification process was monitored using electrochemistry, SEM, XPS. The herein immunosensor demonstrates a good selectivity and high sensitivity against human-cTnI, and is capable of detecting cTnI at concentrations as low as 0.05 ng mL(-1), which is 100 times lower than that possible by conventional methods. It is potential to design the portable sensing platform based on AuNPs and GO nanocomposites for future point-of-care diagnostics.