Microchimica Acta最新文献

Single-level biomarker detection has the limitation of insufficient accuracy in cancer diagnosis. Therefore, the strategy of developing highly sensitive, multi-channel biosensors for high-throughput ctDNA determination is critical to improve the accuracy of early diagnosis of clinical tumors. Herein, in order to achieve efficient detection of up to ten targets for early diagnosis of ovarian cancer, a DNA-nanoswitch-based multi-channel (DNA-NSMC) biosensor was built based on the multi-module catalytic hairpin assembly-mediated signal amplification (CHA) and toehold-mediated DNA strand displacement (TDSD) reaction. Only two different fluorescence signals were used as outputs, combined with modular segmentation strategy of DNA-nanoswitch-based reaction platform; the multi-channel detection of up to ten targets was successfully achieved for the first time. The experimental results suggest that the proposed biosensor is a promising tool for simultaneously detecting multiple biomarkers for the early diagnosis of ovarian cancer, offering new strategies for the early screening, diagnosis, and treatment not only for ovarian cancer but also for other cancers.

Nano- and micro-carriers of therapeutic molecules offer numerous advantages for drug delivery, and the shape of these particles plays a vital role in their biodistribution and their interaction with cells. However, analysing how microparticles are taken up by cells presents methodological challenges. Qualitative methods like microscopy provide detailed imaging but are time-consuming, whereas quantitative methods such as flow cytometry enable high-throughput analysis but struggle to differentiate between internalised and surface-bound particles. Instead, imaging flow cytometry combines the best of both worlds, offering high-resolution imaging with the efficiency of flow cytometry, allowing for quantitative analysis at the single-cell level. This study focuses on fluorescently labelled silicon oxide microchips of various morphologies but related surface areas and volumes: rectangular cuboids and apex-truncated square pyramid microchips fabricated using photolithography techniques, offering a reliable basis for comparison with the more commonly studied spherical particles. Imaging flow cytometry was utilised to evaluate the effect of particle shape on cellular uptake using RAW 264.7 cells and revealed phagocytosis of particles with all shapes. Increasing the particle dose enhanced the uptake, while macrophage stimulation had minimal effect. Using a ratio particle:cell of 10:1 cuboids and spheres showed an uptake rate of approximately 50%, in terms of the percentage of cells with internalised particles, and the average number of particles taken up per cell ranging from about 1-1.5 particle/cell for all the different shapes. This study indicates how differently shaped micro-carriers offer insights into particle uptake variations, demonstrating the potential of non-spherical micro-carriers for precise drug delivery applications.

Macrophages are among the most important components of the innate immune system where the interaction of pathogens and their phagocytosis occur as the first barrier of immunity. When nanomaterials interact with the human body, they have to face macrophages as well. Thus, understanding of nanomaterials-macrophage interactions and underlying mechanisms is crucial. For this purpose, various methods are used. In this study, surface-enhanced Raman scattering (SERS) is proposed by studying lipopolysaccharide (LPS) induced macrophage polarization using gold nanoparticles (AuNPs) as an alternative to the current approaches. For this purpose, the murine macrophage cell line, RAW 264.7 cells, was polarized by LPS, and polarization mechanisms were characterized by nitrite release and reactive oxygen species (ROS) formation and monitored using SERS. The spectral changes were interpreted based on the molecular pathways induced by LPS. Furthermore, polarized macrophages by LPS were exposed to the toxic AuNPs doses to monitor the enhanced phagocytosis and related spectral changes. It was observed that LPS induced macrophage polarization and enhanced AuNP phagocytosis by activated macrophages elucidated clearly from SERS spectra in a label-free non-destructive manner.

Atherosclerosis cardiovascular disease (ASCVD) has become one of the leading death causes in humans. Low-density lipoprotein (LDL) is an important biomarker for assessing ASCVD risk level. Thus, monitoring LDL levels can be an important means for early diagnosis of ASCVD. Herein, a novel electrochemical aptasensor for determination LDL was designed based on nitrogen-doped reduced graphene oxide-hemin-manganese oxide nanoparticles (NrGO-H-Mn₃O₄ NPs) integrated with clustered regularly interspaced short palindromic repeats and associated proteins (CRISPR/Cas12a) system. NrGO-H-Mn₃O₄ NPs not only have a large surface area and remarkable enhanced electrical conductivity but also the interconversion of different valence states of iron in hemin can provide an electrical signal. Nonspecific single-stranded DNA (ssDNA) was bound to NrGO-H-Mn₃O₄ NPs to form a signaling probe and was immobilized on the electrode surface. The CRISPR/Cas12a system has excellent trans-cleavage activity, which can be used to cleave ssDNA, thus detaching the NrGO-H-Mn₃O₄ NPs from the sensing interface and attenuating the electrical signal. Significant signal change triggered by the target was ultimately obtained, thus achieving sensitive detection of the LDL in range from 0.005 to 1000.0 nM with the detection limit of 0.005 nM. The proposed sensor exhibited good stability, selectivity, and stability and achieved reliable detection of LDL in serum samples, demonstrating its promising application prospects for the diagnostic application of LDL.

An intense cathodic electrochemiluminescence (ECL) is reported from a polarized glassy carbon electrode (GCE) in peroxydisulfate solution. After the polarization in 1 M Na₂SO₄ at the potential of - 3.7 V for 3 s, carbon nanosheets (C-NSs) were in situ grown on the surface of the GCE. Measured in 100 mM K₂S₂O₈ solution, the ECL intensity of the GCE/C-NSs is 112-fold that of a bare GCE. The ECL spectrum revealed that the true ECL luminophore in the GCE/C-NSs-peroxydisulfate system is O₂/S₂O₈^2- which is promoted by C-NSs. When Cu²⁺ was electrochemically enriched and reduced to Cu(0) on the catalytic sites of C-NSs, the ECL from GCE/C-NSs/Cu in K₂S₂O₈ solution was decreased with increasing logarithmic concentration of Cu²⁺ in the range from 10 pM to 1 μM, with a limit of detection (LOD) of 3 pM. An immunoanalysis method is proposed via a biometallization strategy using CuS nanoparticles as the tags and carcinoembryonic antigen (CEA) as the model analyte. After the immune recognition in the microplate, the CuS tags in the immunocomplex were dissolved and the resultant Cu²⁺ was electrochemically enriched and reduced on the catalytic sites of C-NSs, quenching the ECL intensity of GCE/C-NSs-O₂/S₂O₈^2- system. The proposed ECL immunoanalysis method was used to quantify CEA in actual serum samples with an LOD of 1.0 fg mL^-1, possessing the advantages of simple electrode modification, high sensitivity and good reproducibility.

Tryptophan(Trp) is being explored as a potential biomarker for various diseases associated with decreased tryptophan levels; however, metabolomic methods are expensive and time-consuming and require extensive sample analysis, making them urgently needed for trace detection. To exploit the properties of Ti₃C₂ MXenes a rational porous methyl orange (MO)-delaminated Ti₃C₂ MXene was prepared via a facile mixing process for the electrocatalytic oxidation of Trp. The hollow-like 3D structure with a more open structure and the synergistic effect of MO and conductive Ti₃C₂ MXene enhanced its electrochemical catalytic capability toward Trp biosensing. More importantly, MO can stabilize Ti₃C₂ MXene nanosheets through noncovalent π-π interactions and hydrogen bonding. Compared with covalent attachment, these non-covalent interactions preserve the electronic conductivity of the Ti₃C₂ MXene nanosheets. Finally, the addition of MO-derived nitrogen (N) and sulfur (S) atoms to Ti₃C₂ MXene enhanced the electronegativity and improved its affinity for specific molecules, resulting in high-performance electrocatalytic activity. The proposed biosensor exhibited a wide linear response in concentration ranges of 0.01-0.3 µM and 0.5-120 µM, with a low detection limit of 15 nM for tryptophan detection, and high anti-interference ability in complex media of human urine and egg white matrices. The exceptional abilities of the MO/Ti₃C₂ nanocatalyst make it a promising electrode material for the detection of important biomolecules.

An electrochemical biosensor based on dual-amplified nucleic acid mode and biocatalytic silver deposition was constructed using catalytic hairpin assembly-hybrid chain reaction (CHA-HCR). The electrochemical detection of silver on the electrode by linear sweep voltammetry (LSV) can be utilized to quantitatively measure miR-205-5p since the amount of silver deposited on the electrode is proportional to the target nucleic acid. The current response values exhibit strong linearity with the logarithm of miR-205-5p concentrations ranging from 0.1 pM to 10 μM, and the detection limit is 28 fM. A consistent trend was found in the results of the qRT-PCR and electrochemical biosensor techniques, which were employed to determine the total RNA recovered from cells, respectively. Moreover, the constructed sensor was used to assess miR-205-5p on various cell counts, and the outcomes demonstrated the excellent analytical efficiency of the proposed strategy. The recoveries ranged from 97.85% to 115.3% with RSDs of 2.251% to 4.869% in human serum samples. Our electrochemical biosensor for miR-205-5p detection exhibits good specificity, high sensitivity, repeatability, and stability. It is a potentially useful sensing platform for tumor diagnosis and tumor type identification in clinical settings.

As a kind of aminoglycoside antibiotics, kanamycin (KAN) is widely applied to animal husbandry and aquaculture. However, the abuse of KAN causes the large-scale discharge of it into rivers, lakes and groundwater, which threatens environmental safety and human health. Therefore, it is imperative to develop a method that is applicable to detect KAN in an efficient and accurate way. The colorimetric method based on enzymes provides a feasible solution for the detection of organic pollutants. However, the extensive application of natural enzymes is constrained by high cost and low stability. Herein, a polyoxometalate-based nanozyme, namely [H₇SiW₉V₃O₄₀(DPA)₃]·4H₂O (SiW₉V₃/DPA) (DPA = dipyridylamine), is synthesized. As a low-cost nanozyme with high stability compared to natural enzymes, SiW₉V₃/DPA performs well in laccase-mimicking activity. It can be used to induce chromogenic reaction between 2,4-dichlorophenol (2,4-DP) and 4-aminoantipyrine (4-AP), which generates red products. With the addition of KAN, the color fades. That is to say, KAN can be detected with colorimetric assay in the concentration range 0.1 to 100 μM with high selectivity and low limit of detection (LOD) of 6.28 μM. Moreover, SiW₉V₃/DPA is applied to KAN detection in lake and river water and milk with satisfactory results. To sum up, polyoxometalate-based nanozyme is expected to provide a promising solution to the detection of organic pollutants in the aquatic environment.

As an ideal transition metal oxide, Co₃O₄ is a P-type semiconductor with excellent electrical conductivity, non-toxicity and low cost. This work reports the successful construction of Co₃O₄ materials derived from metal-organic frameworks (MOFs) using a surfactant micelle template-solvothermal method. The modified electrodes are investigated for their ability to electrochemically detect Pb²⁺ and Cu²⁺ in aqueous environments. By adjusting the mass ratios of alkaline modifiers, the morphological microstructures of Co₃O₄-X exhibit a transition from distinctive microspheres composed of fiber stacks to rods. The results indicate that Co₃O₄-1(NH₄F/CO(NH₂)₂ = 1:0) has a distinctive microsphere structure composed of stacked fibers, unlike the other two materials. Co₃O₄-1/GCE is used as the active material of the modified electrode, it shows the largest peak response currents to Pb²⁺ and Cu²⁺, and efficiently detects Pb²⁺ and Cu²⁺ in the aqueous environment individually and simultaneously. The linear response range of Co₃O₄-1/GCE for the simultaneous detection of Pb²⁺ and Cu²⁺ is 0.5-1.5 μM, with the limits of detection (LOD, S/N = 3) are 9.77 nM and 14.97 nM, respectively. The material exhibits a favorable electrochemical response, via a distinctive Co₃O₄-1 microsphere structure composed of stacked fibers. This structure enhances the number of active adsorption sites on the material, thereby facilitating the adsorption of heavy metal ions (HMIs). The presence of oxygen vacancies (O_V) can also facilitate the adsorption of ions. The Co₃O₄-1/GCE electrode also exhibits excellent anti-interference ability, stability, and repeatability. This is of great practical significance for detecting Pb²⁺ and Cu²⁺ in real water samples and provides a new approach for developing high-performance metal oxide electrochemical sensors derived from MOFs.

A nanohybrid-modified glassy carbon electrode based on conducting polypyrrole doped with carbon quantum dots (QDs) was developed and used for the electrochemical detection of anti-tissue transglutaminase (anti-tTG) antibodies. To improve the polypyrrole conductivity, carrier mobility, and carrier concentration, four types of carbon nanoparticles were tested. Furthermore, a polypyrrole-modified electrode doped with QDs was functionalized with a PAMAM dendrimer and transglutaminase 2 protein by cross-linking with N-hydroxysuccinimide (NHS)/N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride (EDC). The steps of electrode surface modification were surveyed via electrochemical measurements (differential pulse voltammetry (DPV), impedance spectroscopy, and X-ray photoelectron spectroscopy (XPS)). The surface characteristics were observed by scanning electron microscopy (SEM), Fourier transform infrared (FTIR) spectroscopy, and contact angle measurements. The obtained modified electrode exhibited good stability and repeatability. DPV between - 0.1 and 0.6 V (vs. Ag/AgCl 3 M KCl reference electrode) was used to evaluate the electrochemical alterations that occur after the antibody interacts with the antigen (transglutaminase 2 protein), for which the limit of detection was 0.79 U/mL. Without the use of a secondary label, (anti-tTG) antibodies may be detected at low concentrations because of these modified electrode features.