In this paper, we present the first highly porous gold (h-PG) microneedles-based second-generation biosensor for minimally invasive monitoring of glucose in artificial interstitial fluid (ISF). A highly porous microneedles-based electrode was prepared by a simple electrochemical self-templating method that involves two steps, gold electrodeposition and hydrogen bubbling at the electrode, which were realized by applying a potential of −2 V versus a saturated calomel electrode (SCE). The highly porous gold surface of the microneedles was modified by immobilization of 6-(ferrocenyl)hexanethiol (FcSH) as a redox mediator and subsequently by immobilization of a flavin adenine dinucleotide glucose dehydrogenase (FAD-GDH) enzyme using a drop-casting method. The microneedles-based FcSH/FAD-GDH biosensor allows for the detection of glucose in artificial interstitial fluid with an extended linear range (0.1–10 mM), high sensitivity (50.86 µA cm⁻² mM⁻¹), stability (20% signal loss after 30 days), selectivity (only ascorbic acid showed a response about 10% of glucose signal), and a short response time (3 s). These properties were favourably compared to other microneedles-based glucose biosensors reported in the literature. Finally, the microneedle-arrays-based second-generation biosensor for glucose detection was tested in artificial interstitial fluid opportunely spiked with different concentrations of glucose (simulating healthy physiological conditions while fasting and after lunch) and by placing the electrode into a simulated chitosan/agarose hydrogel skin model embedded in the artificial ISF (continuous glucose monitoring). The obtained current signals had a lag-time of about 2 min compared to the experiments in solution, but they fit perfectly into the linearity range of the biosensor (0.1–10 mM). These promising results show that the proposed h-PG microneedles-based sensor could be used as a wearable, disposable, user-friendly, and automated diagnostic tool for diabetes patients.

One of the challenges in the field of biosensors and biofuel cells is to establish a highly efficient electron transfer rate between the active site of redox enzymes and electrodes to fully access the catalytic potential of the biocatalyst and achieve high current densities. We report on very efficient direct electron transfer (DET) between cellobiose dehydrogenase (CDH) from Phanerochaete sordida (PsCDH) and surface modified single walled carbon nanotubes (SWCNT). Sonicated SWCNTs were adsorbed on the top of glassy carbon electrodes and modified with aryl diazonium salts generated in situ from p-aminobenzoic acid and p-phenylenediamine, thus featuring at acidic pH (3.5 and 4.5) negative or positive surface charges. After adsorption of PsCDH, both electrode types showed excellent long-term stability and very efficient DET. The modified electrode presenting p-aminophenyl groups produced a DET current density of 500 μA cm⁻² at 200 mV vs normal hydrogen reference electrode (NHE) in a 5 mM lactose solution buffered at pH 3.5. This is the highest reported DET value so far using a CDH modified electrode and comes close to electrodes using mediated electron transfer. Moreover, the onset of the electrocatalytic current for lactose oxidation started at 70 mV vs NHE, a potential which is 50 mV lower compared to when unmodified SWCNTs were used. This effect potentially reduces the interference by oxidizable matrix components in biosensors and increases the open circuit potential in biofuel cells. The stability of the electrode was greatly increased compared with unmodified but cross-linked SWCNTs electrodes and lost only 15% of the initial current after 50 h of constant potential scanning.

An efficient and inexpensive catalyst for the oxygen reduction reaction (ORR) is the key missing component for large-scale development of fuel cells. Bio-inspired tethered electrocatalysts could be the solution to this problematic reaction. Either unsubstituted Fe phthalocyanine (FePc) or Fe hexadecachloro-phthalocyanine (16(Cl)FePc) was anchored to carbon nanotubes (CNTs) via a pyridine axial ligand. The results show that the fifth coordination plays a major role in increasing the catalytic activity of FePc and 16(Cl)FePc for the ORR. The coordination also allows the decoupling of the metal centre from the carbon support, thus changing the geometrical and electronic structure and hindering the production of H₂O₂. The pentacoordinated catalysts were stable in acidic pH according to the rotating disk analysis, but the activity of the hexadecachloro compound was not higher than that of the unsubstituted phthalocyanine. Cl atoms reduced the coupling between O₂ and Fe, mismatching the energy of the frontier orbitals and lowering the activity towards the reduction of O₂.