Gold Disk Research Articles

A Label-Free Electrochemical Detection of Cadmium Ions Using Aptamer-Based Biosensor

Cadmium is considered one of the most toxic pollutants that can be found in water as well as in soil, which might accumulate in living organisms causing severe effects such as skeletal, cardiovascular, and respiratory diseases. Hence, it is necessary to develop methods allowing for sensitive and fast detection as well as portability. One possibility is the application of biosensors containing aptamer strands as receptor layer selective towards cadmium ions. Here, we present studies on the utilization of DNA aptamer strand for fabrication of sensing layer toward cadmium ions on gold disk macroelectrodes. It is shown that proposed aptasensor enables Cd2+ ions detection in the range from 10 to 50 nM with LOD of 9.5 nM and exhibits high selectivity towards cadmium cations with a response at least two times higher than that for interfering ions. Moreover, studies on the stability revealed that sensing layer preserved its binding properties after storage and allowed for cadmium ions detection in the range from 10 to 50 nM and the aptamer sensing layer could be regenerated and applied for another set of analysis of cadmium ions.

Label-free electrochemical immunosensor for rapid and highly sensitive detection of Brucella abortus in serum

Accurate and efficient detection of Brucella is crucial to control the disease and mitigate its impact on animal health and human safety. In this study, a simple and rapid label-free electrochemical immunosensor for the detection of Brucella abortus was fabricated based on the principle of current decrease after specific capture of the bacteria. A certain proportion of multi-walled carbon nanotubes (MWCNTs), dopamine hydrochloride (DA), and gold nanoparticles (AuNPs) were modified on the surface of a gold disc electrode. Then, Brucella abortus antibody and bovine serum albumin (BSA) were immobilized in sequence to complete the assembly of the sensor. The electrodes from various assembly steps were characterized by a cyclic voltammetry (CV) curve. The immunosensor exhibited high sensitivity with a wide linear range between 10 CFU/mL and 108 CFU/mL. Additionally, the sensor displayed good repeatability, stability, and interference resistance. The recovery rates for low, medium, and high concentrations of bacteria were all within the range of 90 % to 100 %, indicating a high level of accuracy. The method established in this study can provide reference and assistance for early diagnosis of Brucella abortus in clinical practice.

Graphene-assisted metamaterial absorber for refractive index sensor at lower optical frequencies

Numerical studies on a graphene-assisted metamaterial absorber operating at lower optical frequencies are presented. The metamaterial absorber comprises one-dimensional gold grating with a gold disk array in grooves on a monolayer graphene supported by a glass substrate. Numerical simulations are conducted using Computer Simulation Technology Microwave Studio to investigate the underlying absorption mechanisms in the system. The absorber shows a very sharp absorption band with almost unity absorptance and nearly 20 nm full width at half maximum at a wavelength of 766 nm. The simulated absorption spectrum of the absorber is theoretically validated using coupled mode theory, showing close agreement with the theoretical predictions. The absorption occurs due to the simultaneous excitation of localized and propagating surface plasmons under the transverse electric mode (where the electric field is aligned parallel to the length of the grating ridge) excitation in the system. The sensing performance of the absorber has been evaluated, demonstrating a sensitivity as high as 667 nm/RIU. Due to the robustness of the design, this absorber can be used to sense biological, chemical, gaseous, and even acidic samples since it is not comprised of any photoresist. The absorber holds potential for diverse applications, including graphene-based devices, narrowband filters, thermal emitters, solar cells, imaging apparatus, etc.

Design of a Rotating Disk Electrode setup operating under high pressure and temperature: Application to CO2 reduction on gold

We describe the design and development of a rotating disk electrode (RDE) cell capable of operating at pressures up to 200 bar and temperatures up to 200 °C. This setup enables electrochemical surface characterization through techniques such as voltammetry and impedance spectroscopy, under different mass transport regimes. Furthermore, evaluation of catalytic performance, including CO2 reduction, is possible as the system works in a semi-continuous mode interfaced with online gas sample measurements. As a proof of concept of the high-pressure cell designed, we examined the temperature-dependent changes in the cyclic voltammograms (CVs) of polycrystalline gold up to 150 °C and 50 bar. Additionally, online catalytic performance of CO2 reduction to CO on a rotating polycrystalline gold disk electrode was investigated under different pressure and temperature. Our results indicate a positive impact of temperature on the faradaic efficiency (FE) towards CO up to 50 °C, beyond which a rapid drop in performance was observed at atmospheric pressure. Conversely, increasing pressure positively affected CO2 solubility in the electrolyte, resulting in enhanced FE towards CO, reaching approximately 90 % at 6 bar compared to 40 % at atmospheric pressure. Notably, further increases in pressure did not significantly alter the FE, but led to higher current densities. Moreover, at pressures exceeding 6 bar, we observed a plateau in efficiency at temperatures higher than 50 °C. This observation suggests that increasing pressure can sustain CO2 electrolysis, validating the hypothesis that increasing CO2 solubility would suppress catalytic decay at higher temperatures. This study opens up promising avenues for future investigations in electrocatalysis, ranging from fundamental explorations of surface modifications induced by variations in temperature and pressure to the development of high-performance catalysts.

Honeycomb gold specimen supports enabling orthogonal focussed ion beam-milling of elongated cells for cryo-ET

Cryo-focussed ion beam (FIB)-milling is a powerful technique that opens up thick, cellular specimens to high-resolution structural analysis by electron cryotomography (cryo-ET). FIB-milled lamellae can be produced from cells on grids, or cut from thicker, high-pressure frozen specimens. However, these approaches can put geometrical constraints on the specimen that may be unhelpful, particularly when imaging structures within the cell that have a very defined orientation. For example, plunge frozen rod-shaped bacteria orient parallel to the plane of the grid, yet the Z-ring, a filamentous structure of the tubulin-like protein FtsZ and the key organiser of bacterial division, runs around the circumference of the cell such that it is perpendicular to the imaging plane. It is therefore difficult or impractical to image many complete rings with current technologies. To circumvent this problem, we have fabricated monolithic gold specimen supports with a regular array of cylindrical wells in a honeycomb geometry, which trap bacteria in a vertical orientation. These supports, which we call “honeycomb gold discs”, replace standard EM grids and when combined with FIB-milling enable the production of lamellae containing cross-sections through cells. The resulting lamellae are more stable and resistant to breakage and charging than conventional lamellae. The design of the honeycomb discs can be modified according to need and so will also enable cryo-ET and cryo-EM imaging of other specimens in otherwise difficult to obtain orientations.

The 2.5th generation enzymatic sensors based on the construction of quasi-direct electron transfer type NAD(P)-Dependent dehydrogenases

We introduce a versatile method to convert NAD+ or NADP+ -dependent dehydrogenases into quasi-direct electron transfer (quasi-DET)-type dehydrogenases, by modifying with a mediator on the enzyme surface toward the development of 2.5th generation enzymatic sensors. In this study, we use β-hydroxybutyrate (BHB) dehydrogenase (BHBDh) from Alcaligenes faecalis (AfBHBDh) as a representative NAD+ or NADP+ -dependent dehydrogenase. BHBDhs are important in ketone monitoring, especially for the diagnosis of diabetic ketoacidosis. We modified AfBHBDh with a thiol-reactive phenazine ethosulfate (trPES). We designed, constructed, and modified mutant BHBDhs harboring cysteine residues within 20 Å from the C4 nicotinamide in NAD+/NADH. Mutants Ser65Cys, Thr96Cys, and Lys106Cys showed indistinguishable catalytic activities from the wild-type enzyme, even after trPES modification. These trPES-modified mutants were immobilized on gold disk electrodes via amine coupling with succinimide-groups of dithiobis (succinimidyl hexanoate) self-assembled monolayers for electrochemical measurements. Considering there is a wide range of BHB concentrations, we exploited the linear regression in log scales. The linear range for the sensors with trPES-modified BHBDh mutants Ser65Cys, Thr96Cys, and Lys106Cys were 0.1–4.0 mM in both buffer solution and artificial interstitial fluid (ISF). They have limits of detection of 0.047 mM for Ser65Cys, 0.15 mM for Thr96Cys, and 0.060 mM for Lys106Cys in buffer solution, and 0.12 mM, 0.089 mM, and 0.044 mM in artificial ISF, respectively. These results indicate that redox mediator modification of NAD(P)-dependent dehydrogenases converts them into quasi-DET-type dehydrogenases, thereby enabling their utilization in 2.5th generation enzymatic sensors, which will facilitate the construction of enzymatic sensors suitable for continuous monitoring systems.

Electrochemical detection of manganese ions using aptamer-based layers

Heavy metals are one of the major pollutants found in drinking water and their abnormal level may pose a threat to human's health and life. Manganese also belongs to heavy metals group, and it is generally used in production of batteries, fertilizers, and ceramics. Even though, Mn is necessary for proper development of central nervous system, its elevated concentration might lead to certain diseases such as epilepsies, cell death in focal cerebral ischemia as well as neurodegenerative diseases such as Huntington and Alzheimer. Hence, it is crucial to elaborate novel methods for manganese ions detection that could be applied for in situ analysis of water samples.Herein, we present the studies on the electrochemical detection of manganese ions using aptamer-modified electrodes. This is the first attempt of application of aptamer strands as receptor layers for electrochemical analysis of manganese ions and for that purpose gold disk electrodes served as transducers, which were further modified with disulfide - based aptamers and 6-mercapto-1-hexanol blocking agent. The electrochemical measurements concerned the choice of the conditions for formation of aptamer receptor layer as well as the type of redox indicator that served as the source of current signal. The studies referred to the definition of aptasensor working parameters including the verification of the possibility of manganese ion detection in cell culture media. It was shown that it was possible to detect Mn2+ ions within 25 nM–1 μM concentration and the proposed aptasensor exhibited high selectivity towards target analyte for which at least 2 – times higher response was recorded than for interfering ions. Moreover, the possibility of Mn2+ detection in real samples was depicted followed by stability and regeneration studies.

Toward Complete Optical Coupling to Confined Surface Polaritons.

Optical coupling between propagating light and confined surface polaritons plays a pivotal role in the practical design of nanophotonic devices. However, the coupling efficiency decreases dramatically with the degree of mode confinement due to the mismatch that exists between the light and polariton wavelengths, and despite the intense efforts made to explore different mechanisms proposed to circumvent this problem, the realization of a flexible scheme to efficiently couple light to polaritons remains a challenge. Here, we experimentally demonstrate an efficient coupling of light to surface-plasmon polaritons assisted by engineered dipolar scatterers placed at an optimum distance from the surface. Specifically, we fabricate gold disks separated by a silica spacer from a planar gold surface and seek to achieve perfect coupling conditions by tuning the spacer thickness for a given scatterer geometry that resonates at a designated optical frequency. We measure a maximum light-to-plasmon coupling cross section of the order of the square of the light wavelength at an optimum distance that results from the interplay between a large particle-surface interaction and a small degree of surface-driven particle-dipole quenching, both of which are favored at small separations. Our experiments, in agreement with both analytical theory and electromagnetic simulations, support the use of optimally placed engineered scatterers as a disruptive approach to solving the long-standing problem of in/out-coupling in nanophotonics.

Studies on the Aptasensor Miniaturization for Electrochemical Detection of Lead Ions.

Lead poses severe effects on living organisms, and since Pb2+ ions tend to accumulate in different organs, it is crucial to monitor Pb2+ concentration in samples such as water and soil. One of the approaches is the utilization of biosensors combined with aptamer-based layers for the electrochemical detection of lead ions. Herein, we present the studies of applying miniaturized screen-printed transducers as solid surfaces to fabricate aptamer layers. As the research is the direct continuation of our previous studies regarding the use of gold disk electrodes, the working parameters of elaborated aptasensors were defined, including the range of linear response (10-100 nM), selectivity as well as stability, regeneration, and feasibility of application for the analysis of real samples. This was achieved using voltammetric techniques including cyclic and square-wave voltammetry in the presence of methylene blue redox indicator.

Task-based detectability in anatomical background in digital mammography, digital breast tomosynthesis and synthetic mammography

Objective. Determining the detectability of targets for the different imaging modalities in mammography in the presence of anatomical background noise is challenging. This work proposes a method to compare the image quality and detectability of targets in digital mammography (DM), digital breast tomosynthesis (DBT) and synthetic mammography. Approach. The low-frequency structured noise produced by a water phantom with acrylic spheres was used to simulate anatomical background noise for the different types of images. A method was developed to apply the non-prewhitening observer model with eye filter (NPWE) in these conditions. A homogeneous poly(methyl) methacrylate phantom with a 0.2 mm thick aluminium disc was used to calculate 2D in-plane modulation transfer function (MTF), noise power spectrum (NPS), noise equivalent quanta, and system detective quantum efficiency for 30, 50 and 70 mm thicknesses. The in-depth MTFs of DBT volumes were determined using a thin tungsten wire. The MTF, system NPS and anatomical NPS were used in the NPWE model to calculate the threshold gold thickness of the gold discs contained in the CDMAM phantom, which was taken as reference. Main results. The correspondence between the NPWE model and the CDMAM phantom (linear Pearson correlation 0.980) yielded a threshold detectability index that was used to determine the threshold diameter of spherical microcalcifications and masses. DBT imaging improved the detection of masses, which depended mostly on the reduction of anatomical background noise. Conversely, DM images yielded the best detection of microcalcification s. Significance. The method presented in this study was able to quantify image quality and object detectability for the different imaging modalities and levels of anatomical background noise.

Gold film deposition by infrared laser photothermal treatment on 3D-printed electrodes: electrochemical performance enhancement and application.

3D printing has attracted the interest of researchers due to its creative freedom, low cost, and ease of operation. Because of these features, this technology has produced different types of electroanalytical platforms. Despite their popularity, the thermoplastic composites used for electrode fabrication typically have high electrical resistance, resulting in devices with poor electrochemical performance. Herein, we propose a new strategy to improve the electrochemical performance of 3D-printed electrodes and to gain chemical selectivity towards glucose detection. The approach involves synthesising a nanostructured gold film using an infrared laser source directly on the surface of low-contact resistance 3D-printed electrodes. The laser parameters, such as power, focal distance, and beam scan rate, were carefully optimised for the modification steps. Scanning electronic microscopy and energy-dispersive X-ray spectroscopy confirmed the morphology and composition of the nanostructured gold film. After modification, the resulting electrodes were able to selectively detect glucose, encouraging their use for sensing applications. When compared with a gold disc electrode, the gold-modified 3D-printed electrode provided a 44-fold current increase for glucose oxidation. As proof of concept, the devices were utilised for the non-enzymatic catalytic determination of glucose in drink samples, demonstrating the gold film's catalytic nature and confirming the analytical applicability with more precise results than commercial glucometers.

Label-free and ultrasensitive electrochemical detection of nucleic acids based on an exonuclease III-assisted target recycling amplification strategy using a heated gold disk electrode.

The present work demonstrates a label-free, rapid and ultrasensitive electrochemical sensor for specific DNA detection with an exonuclease III (Exo III)-assisted target recycling amplification strategy and elevated electrode temperature at a heated gold disk electrode (HAuDE). The proposed electrochemical DNA (E-DNA) sensor was designed such that in the presence of the target DNA, the electrode self-assembled capture probe hybridizes with the target DNA to form a duplex structure, which triggers Exo III to specifically recognize this structure and selectively digest the capture probe, while the released target DNA underwent recycling to hybridize with a new capture probe, leading to the gradual digestion of a large amount of capture probes. It was found that during the digestion period, the activity of Exo III could be significantly improved by elevating electrode temperature, thus promoting the digestion reaction and improving the sensitivity for target DNA detection. Furthermore, an electrochemical indicator ([Ru(NH3)6]3+) was electrostatically bound to the capture probe, leading to a significant square wave voltammetry (SWV) response, which directly related to the amount and length of the capture probes remaining in the electrode and provided a quantitative measure for target DNA detection. The proposed strategy realized the highly sensitive detection of the target DNA with a detection limit of 26 aM (S/N = 3) at an electrode temperature of 40 °C during the digestion period, which was about two magnitudes lower than that at 24 °C.

Synthesis and Some Electrochemical Properties of Carbazole Derivative Monomers for Optoelectronic Device Design

Low-cost instruments are used to achieve high sensitivity, accuracy and precision with electrochemical techniques, making them the most suitable analytical methods used to investigate the electrochemical properties of a new molecule. Among these techniques, cyclic voltammetry, which is widely employed, allows for the determination of electronic characteristics such as molecular energy levels and orbital structure, redox properties, sensor capabilities, surface activity, and more. The carbazole molecule can undergo derivatization from various positions, allowing for alterations in its electrical and optical properties. These compounds serve as heterocyclic building blocks that can be utilized as materials for organic sensitizers and semiconductors in optoelectronic devices. Due to the presence of a hydrogen atom in the nitrogen-hydrogen (N-H) bond within the carbazole structure, which can be replaced with different functional groups, carbazole is highly suitable for nitrogen-based derivatization studies. In this study, two different carbazole monomers (IIa and IIIa), which could be potential optoelectronic, were synthesized using the Ullman and Suzuki-Miyaura reaction and characterized using 1H NMR, 13C NMR, UV-Vis, and Flouresence spectroscopy techniques. The cyclic voltammetry experiments were performed using three different working electrodes (Au, Gc, Pt disk electrodes) for each compound. Since the optimum oxidation of the compounds was obtained from the voltammograms on the gold disk electrode, this electrode was used in the calculation of energy levels. HOMO and Eg values of the compounds were deduced from cyclic voltammetry experiments and the optical absorption bands, respectively.

Bacteriophage long tail fibre proteins as a biorecognition element in electrochemical biosensors for the detection of Salmonella in lake water

We developed an electrochemical impedance spectroscopy (EIS) biosensor based on bacteriophage long tail fibre proteins (LTFPs) to capture and detect Salmonella. First, based on the genetic information of a Salmonella enterica bacteriophage (SEP37), we found that the genes ORF99, ORF100, and ORF256 encode proteins associated with host recognition. We used the pETdue1 plasmid to construct ORF99–ORF100 and ORF256 co-expression vectors to express (LTFPs) in Escherichia coli and used purified LTFPs as specific biorecognition elements. Then, we fixed gold nanoparticles (AuNPs), cysteamine (Cys), and LTFPs on a gold disk electrode surface via layer-by-layer assembly to construct an EIS biosensor. Salmonella in the sample can be captured by LTFPs on the electrode surface and reduce the electron transfer sites. We applied the EIS biosensor to detect Salmonella spiked in lake water. It showed a good linear relationship within the concentration range of 10 ∼ 1 × 106 colony-forming units (CFU)/mL. The detection limit was 9 CFU/mL. This method based on bacteriophage LTFPs does not require complex pretreatment steps and extends the application of tail fibre proteins as a biorecognition element in the detection field.

Photoluminescence Enhancement of Monolayer WS2 by n-Doping with an Optically Excited Gold Disk.

In nanophotonics and quantum optics, we aim to control and manipulate light with tailored nanoscale structures. Hybrid systems of nanostructures and atomically thin materials are of interest here, as they offer rich physics and versatility due to the interaction between photons, plasmons, phonons, and excitons. In this study, we explore the optical and electronic properties of a hybrid system, a naturally n-doped monolayer WS2 covering a gold disk. We demonstrate that the nonresonant excitation of the gold disk in the high absorption regime efficiently generates hot carriers via localized surface plasmon excitation, which n-dope the monolayer WS2 and enhance the photoluminescence emission by regulating the multiexciton population and stabilizing the neutral exciton emission. The results are relevant to the further development of nanotransistors in photonic circuits and optoelectronic applications.

The cyanidation of sub microscopic gold: An experimental and molecular modeling study

Sub microscopic refractory gold ore is an important resource of gold with the depletion of free-milling gold deposits in this century. It has been generally accepted that submicroscopic gold mainly in the form of solid-solution and colloidal particles are physically locked inside mineral phases and impervious to cyanide solution. However, some researches on synthetic nanoparticles show they have very different physical and chemical properties which can affect their dissolution behavior.This research has been conducted to investigate the dissolution behavior of gold sub microscopic particles experimentally and theoretically using density functional theory. For this purpose, 20 nm gold particles have been synthesized and dissolved in cyanide solution under controlled condition. The rate of dissolution has been monitored during cyanidation and compared with a gold disk. The results depict the rate of dissolution per surface area is significantly reduced in gold nanoparticles (30 times, the surface ratio of 14/9 m2/g and 0.006 m2/g in samples is 2500 times smaller while the dissolution rate in nanoparticles is 83 times faster). Also, the smaller nanoparticles have much slower dissolution kinetics than large nanoparticles (5.485 × 10−5 and 4.280 × 10−6 g/m2.s for 16 and 2 nm particles, respectively). Two different dissolution regimes can be identified for >3 nm and < 3 nm nanoparticles. In larger particles, by increasing the number of surface atoms with weaker bond energy and also lower oxidation potential, the dissolution rate increases compared to bulk gold. In nanoparticles smaller than 3 nm, the electrochemical reaction in the cyanide medium is disrupted by the reduction of oxidation potential and decreasing the conductivity as the requirement for electrochemical dissolution of gold in cyanide.The molecular modeling study on the gold clusters of 1 to 20 atom confirm that the oxidation of small clusters of gold is more difficult than bulk gold due to the high ionization energy (9.66 vs. 5.4 eV). Also, the monoatomic gold has a much weaker interaction with cyanide (−4.36 vs. -1.6 eV) and higher interaction with oxygen (−0.64 vs. -0.15 eV) compared to the bulk gold. The HOMO of monoatomic and bulk gold (−9.22 and − 4.5 eV) as well as LUMO of cyanide and oxygen (1.734 and − 4.657 eV), as two reactants in cyanidation process confirm the results based on HSAB theory. As a result, oxygen competes with cyanide for adsorption on the surface of gold nanoparticles, disrupting the first step of electrochemical dissolution of gold, as the adsorption of cyanide and the formation of the Au-CN− complex. This is possible by deactivating the surface by creating a resistant layer of gold oxide or electrostatic adsorption of oxygen on the gold surface and passivating the surface. The modeling data are very well in agreement with the experimental results indicating the inherent inactivity of free surface gold sub microscopic particles during the cyanidation process.

Salt-induced nanoporous gold electrodes: One-step scalable fabrication of high-performance flexible electrochemical sensors from gold nanoparticles

Here, a simple filtration method for the fabrication of salt-induced nanoporous gold electrodes (SINPGEs) from gold nanoparticles (AuNPs) is reported. This method employs sodium chloride (NaCl) to trigger the aggregation, room-temperature sintering and region-selective deposition of AuNPs on patterned polyvinylidene fluoride (PVDF) membranes during filtration, which enables the one-step scalable fabrication of flexible SINPGEs three-electrode arrays with high conductivity, good mechanical property, large surface area and extremely low cost. Moreover, compared with traditional gold disc electrodes, the nanoporous structure of SINPGEs was demonstrated to apparently improve the electrochemical responses of various electroactive species, e.g., dopamine (DA), uric acid (UA), ascorbic acid (AA) and hydrogen peroxide (H2O2), reflected by the apparent enlargement of redox currents. In addition, the passivation effect of chloride ions (Cl−) on the oxidation of H2O2 at SINPGEs can be greatly repressed by a simple electrochemical activation approach, producing a sensitive H2O2 sensor with a calibration range of 1.0–1000 μM in the presence of 0.1 M Cl−. The present work thus establishes a new approach to the scalable construction of gold electrode-based electrochemical sensors with high performance and low cost.

Plasmonic gold disk nanoarrays-based surface lattice resonance sensor with high figure of merit and its near-field and far-field coupling characteristics

In the field of localized surface plasmon resonance sensing, narrow linewidth, high-quality factor, and high absorptivity have always been the goals that researchers are committed to achieving. The primary methods for achieving this goal are near-field coupling and far-field coupling. Due to the limitations of nanomanufacturing processes, current research often only focuses on single coupling methods such as far-field coupling or near-field coupling, with few relevant reports combining the two coupling methods. Based on this, this paper proposes a single-layer gold nano-disk array structure, which is simple and easy to manufacture. Through modal field analysis and structural parameter adjustment, some significant results have been achieved. On the one hand, the suggested gold nano-disk array structure yields a resonant peak based on far-field coupling with a full width at half maximum (FWHM) of less than 0.05 nm and a figure of merit of more than 15 000 (the quality factor is more than 20 000). On the other hand, it has been successfully accomplished to combine near-field coupling and far-field coupling, leading to a composite coupling peak that incorporates the benefits of both types of coupling. Moreover, this coupling peak has excellent characteristics such as narrow FWHM (less than 3 nm) and high absorptivity (up to 80.5%), which is superior to most reported sensing structures and more in line with practical sensing application requirements. In this paper, the potential of the gold disk array structure as a high-performance sensing structure is further explored, providing reference and new ideas for the design of easy-to-prepare and high-performance electromagnetic resonance devices.

Heating promoted super sensitive electrochemical detection of p53 gene based on alkaline phosphatase and nicking endonuclease Nt.BstNBI-assisted target recycling amplification strategy at heated gold disk electrode

An ultrasensitive electrochemical biosensor for detecting p53 gene was fabricated based on heated gold disk electrode coupling with endonuclease Nt.BstNBI-assisted target recycle amplification and alkaline phosphatase (ALP)-based electrocatalytic signal amplification. For biosensor assembling, biotinylated ssDNA capture probes were first immobilized on heated Au disk electrode (HAuDE), then combined with streptavidin-alkaline phosphatase (SA-ALP) by biotin-SA interaction. ALP could catalyze the hydrolysis of ascorbic acid 2-phosphate (AAP) to produce ascorbic acid (AA). While AA could induce the redox cycling to generate electrocatalytic oxidation current in the presence of ferrocene methanol (FcM). When capture probes hybridized with p53, Nt.BstNBI would recognize and cleave the duplexes and p53 was released for recycling. Meanwhile, the biotin group dropt from the electrode surface and subsequently SA-ALP could not adhere to the electrode. The signal difference before and after cleavage was proportional to the p53 gene concentration. Furthermore, with electrode temperature elevated, the Nt.BstNBI and ALP activities could be increased, greatly improving the sensitivity and efficiency for p53 detection. A detection limit of 9.5 × 10−17 M could be obtained (S/N = 3) with an electrode temperature of 40 °C, ca. four magnitudes lower than that at 25 °C.

Evaluation of Human Gingival Fibroblasts (HGFs) Behavior on Innovative Laser Colored Titanium Surfaces.

The use of ytterbium laser to obtain colored titanium surfaces is a suitable strategy to improve the aesthetic soft tissue results and reduce implant failures in oral rehabilitation. To investigate the relationship between novel laser-colored surfaces and peri-implant soft tissues, Human Gingival Fibroblasts (HGFs) were cultured onto 12 colored titanium grade 1 light fuchsia, dark fuchsia, light gold, and dark gold disks and their viability (MTT Assay), cytotoxicity (lactate dehydrogenase release), and collagen I secretion were compared to the machined surface used as control. Optical and electronic microscopies showed a HGF growth directly correlated to the roughness and wettability of the colored surfaces. A higher viability percentage on dark fuchsia (125%) light gold (122%), and dark gold (119%) samples with respect to the machined surface (100%) was recorded. All specimens showed a statistically significant reduction of LDH release compared to the machined surface. Additionally, a higher collagen type I secretion, responsible for an improved adhesion process, in light fuchsia (3.95 μg/mL) and dark gold (3.61 μg/mL) compared to the machined surface (3.59 μg) was recorded. The in vitro results confirmed the innovative physical titanium improvements due to laser treatment and represent interesting perspectives of innovation in order to ameliorate aesthetic dental implant performance and to obtain more predictable osteo and perio-osteointegration long term implant prognosis.