Influence of enzyme immobilization and skin-sensor interface on non-invasive glucose determination from interstitial fluid obtained by magnetohydrodynamic extraction.

We integrated a magnetohydrodynamic fluid extractor with an amperometric glucose biosensor to develop a wearable device for non-invasive glucose monitoring. Reproducible fluid extraction through the skin and efficient transport of the extracted fluid to the biosensor surface are prerequisites for non-invasive glucose monitoring. We optimized the enzyme immobilization and the interface layer between the sensing device and the skin. The monitoring device was evaluated by extracting fluid through porcine skin followed by glucose detection at the biosensor. The biosensor featured a screen-printed layer of Prussian Blue that was coated with a layer containing glucose oxidase. Both physical entrapment of glucose oxidase in chitosan and tethering of glucose oxidase to electrospun nanofibers were evaluated. Binding of glucose oxidase to nanofibers under mild conditions provided a stable biosensor with analytical performance suitable for accurate detection of micromolar concentrations of glucose. Hydrogels of varying thickness (95-2000 µm) as well as a thin (30 µm) nanofibrous polycaprolactone mat were studied as an interface layer between the biosensor and the skin. The effect of mass transfer phenomena at the biosensor-skin interface on the analytical performance of the biosensor was evaluated. The sensing device detected glucose extracted through porcine skin with an apparent (overall) sensitivity of -0.8 mA/(M·cm2), compared to a sensitivity of -17 mA/(M·cm2) for measurement in solution. The amperometric response of the biosensor correlated with the glucose concentration in the fluid that had been extracted through porcine skin with the magnetohydrodynamic technique.

Integrating Carbon Nanomaterials with Metals for Bio-sensing Applications.

Age structure in most developed countries is changing fast as the average lifespan is increasing significantly, calling for solutions to provide improved treatments for age-related neurological diseases and disorders. In order to address these problems, a reliable way of recording information about neurotransmitters from in vitro and in vivo applications is needed to better understand neurological diseases and disorders as well as currently used treatments. Likewise, recent developments in medicine, especially with the opioid crisis, are demanding a swift move to personalized medicine to administer patient needs rather than population-wide averages. In order to enable the so-called personalized medicine, it is necessary to be able to do measurements in vivo and in real time. These actions require sensitive and selective detection of different analytes from very demanding environments. Current state-of-the-art materials are unable to provide sensitive and selective detection of neurotransmitters as well as the required time resolution needed for drug molecules at a reasonable cost. To meet these challenges, we have utilized different metals to grow carbon nanomaterials and applied them for sensing applications showing that there are clear differences in their electrochemical properties based on the selected catalyst metal. Additionally, we have combined atomistic simulations to support optimizing materials for experiments and to gain further understanding of the atomistic level reactions between different analytes and the sensor surface. With carbon nanostructures grown from Ni and Al + Co + Fe hybrid, we can detect dopamine, ascorbic acid, and uric acid simultaneously. On the other hand, nanostructures grown from platinum provide a feasible platform for detection of H2O2 making them suitable candidates for enzymatic biosensors for detection of glutamate, for example. Tetrahedral amorphous carbon electrodes have an ability to detect morphine, paracetamol, tramadol, and O-desmethyltramadol. With carbon nanomaterial-based sensors, it is possible to reach metal-like properties in sensing applications using only a fraction of the metal as seed for the material growth. We have also seen that by using nanodiamonds as growth catalyst for carbon nanofibers, it is not possible to detect dopamine and ascorbic acid simultaneously, although the morphology of the resulting nanofibers is similar to the ones grown using Ni. This further indicates the importance of the metal selection for specific applications. However, Ni as a continuous layer or as separate islands does not provide adequate performance. Thus, it appears that metal nanoparticles combined with fiber-like morphology are needed for optimized sensor performance for neurotransmitter detection. This opens up a new research approach of application-specific nanomaterials, where carefully selected metals are integrated with carbon nanomaterials to match the needs of the sensing application in question.

Unmodified and multi-walled carbon nanotube modified tetrahedral amorphous carbon (ta-C) films as in vivo sensor materials for sensitive and selective detection of dopamine.

Unmodified and multi-walled carbon nanotube (MWCNT) modified tetrahedral amorphous carbon (ta-C) films of 15 and 50 nm were investigated as potential in vivo sensor materials for the detection of dopamine (DA) in the presence of the main interferents, ascorbic acid (AA) and uric acid (UA). The MWCNTs were grown directly on ta-C by chemical vapor deposition (designated as ta-C+CNT) and were characterized with X-ray photoelectron spectroscopy, Raman spectroscopy, scanning and transmission electron microscopy. Electroanalytical sensitivity and selectivity were determined with cyclic voltammetry. Biocompatibility of the materials was assessed with cell cultures of mouse neural stem cells (mNSCs). The detection limits of DA for both ta-C and ta-C+CNT electrodes ranged from 40 to 85 nM, which are well within the required range for in vivo detection. The detection limits were lower for both ta-C and ta-C+CNT electrodes with 50 nm of ta-C compared to 15 nm. The ta-C electrodes showed a large dynamic linear range of 0.01-100 µM but could not resolve between the oxidation peaks of DA, AA and UA. Modification with MWCNTs, however, resulted in excellent selectivity and all three analytes could be detected simultaneously at physiologically relevant concentrations using cyclic voltammetry. Based on cell culture of mNSCs, both ta-C and ta-C+CNT exhibited good biocompatibility, demonstrating their potential as in vivo sensor materials for the detection of DA.

Electrochemical Fouling of Dopamine and Recovery of Carbon Electrodes.

A significant problem with implantable sensors is electrode fouling, which has been proposed as the main reason for biosensor failures in vivo. Electrochemical fouling is typical for dopamine (DA) as its oxidation products are very reactive and the resulting polydopamine has a robust adhesion capability to virtually all types of surfaces. The degree of DA fouling of different carbon electrodes with different terminations was determined using cyclic voltammetry (CV) and scanning electrochemical microscopy (SECM) approach curves and imaging. The rate of electron transfer kinetics at the fouled electrode surface was determined from SECM approach curves, allowing a comparison of insulating film thickness for the different terminations. SECM imaging allowed the determination of different morphologies, such as continuous layers or islands, of insulating material. We show that heterogeneous modification of carbon electrodes with carboxyl-amine functionalities offers protection against formation of an insulating polydopamine layer, while retaining the ability to detect DA. The benefits of the heterogeneous termination are proposed to be due to the electrostatic repulsion between amino-functionalities and DA. Furthermore, we show that the conductivity of the surfaces as well as the response toward DA was recovered close to the original performance level after cleaning the surfaces for 10-20 cycles in H2SO4 on all materials but pyrolytic carbon (PyC). The recovery capacity of the PyC electrode was lower, possibly due to stronger adsorption of DA on the surface.

Integrated Carbon Nanostructures for Detection of Neurotransmitters.

Carbon-based materials, such as diamond-like carbon (DLC), carbon nanofibers (CNFs), and carbon nanotubes (CNTs), are inherently interesting for neurotransmitter detection due to their good biocompatibility, low cost and relatively simple synthesis. In this paper, we report on new carbon-hybrid materials, where either CNTs or CNFs are directly grown on top of tetrahedral amorphous carbon (ta-C). We show that these hybrid materials have electrochemical properties that not only combine the best characteristics of the individual "building blocks" but their synergy makes the electrode performance superior compared to conventional carbon based electrodes. By combining ta-C with CNTs, we were able to realize electrode materials that show wide and stable water window, almost reversible electron transfer properties and high sensitivity and selectivity for detecting dopamine in the presence of ascorbic acid. Furthermore, the sensitivity of ta-C + CNF hybrids towards dopamine as well as glutamate has been found excellent paving the road for actual in vivo measurements. The wide and stable water window of these sensors enables detection of other neurotransmitters besides DA as well as capability of withstanding higher potentials without suffering from oxygen and hydrogen evolution.

Improving the function of dopamine electrodes with novel carbon materials.

For therapeutic purposes, an accurate measurement of dopamine level in situ would be highly desirable. A novel strategy for the selective determination of dopamine concentration based on the diamond-like carbon (DLC) electrode is presented in this abstract. The developed DLC electrode is able to detect 10 µM dopamine and has improved sensitivity compared to platinum. Compared to carbon fiber electrodes, the DLC electrode is more stable because the background current is much lower.