Tuning the Probe-Bilayer Architecture of Silver Nanoneedle-based Ion Channel Probes.

Ion channel probes, as one of the ion channel platforms, provide an appealing opportunity to perform localized detection with a high precision level. These probes come basically in two classes: glass and metal. While the glass-based probes showed the potential to be employed for molecular sensing and chemical imaging, these probes still suffer from limited resolution and lack of control over protein insertion. On the other hand, metal-based nanoneedle probes (gold and silver) have been recently developed to allow reducing probe dimensions to the nanoscale geometry. More specifically, silver probes are preferable owing to their ability to mitigate the channel current decay observed with gold probes and provide a stable DC channel current. However, there are still some challenges related to the probe design and bilayer curvature that render such probes insensitive to small changes in the tip-substrate distance. Herein, we introduce two main pathways to control the probe-bilayer architecture; the first is by altering the probe shape and geometry during the fabrication process of silver probes. The second pathway is by altering the surface characteristics of the silver probe via an electrophoretic deposition process. Our findings reveal that varying the electrochemical etching parameters results in different probe geometries and producing sharper tips with a 2-fold diameter reduction. In addition, the electrophoretic deposition of a cathodic paint on the silver nanoneedle surface led to a miniaturized exposed silver tip that enables the formation of a confined bilayer. We further investigated the characteristics of bilayers supported on both the sharper nanoneedles and the HSR-coated silver probes produced by controlling the etching conditions and electrodeposition process, respectively. We believe this work paves the way to rationally design silver nanoneedle ion channel probes, which are well suited for localized molecular sensing and chemical imaging.

Maintaining Single-Channel Recordings on a Silver Nanoneedle through Probe Design and Feedback Tip Positioning Control.

Ion channel proteins showed great promise in the field of nanopore sensing and molecular flux imaging applications due to the atomic-level precision of the pore size and a high signal-to-noise ratio. More specifically, ion channel probes, where the protein channels are integrated at the end of a solid probe, can achieve highly localized detection. Metal probe materials such as gold and silver have been developed to support lipid bilayers and enable the use of smaller probes, or nanoneedles, compared to more traditional glass micropipette ion channel probes. Silver probes are preferable because they support sustained DC stable channel current due to the AgCl layer formed around the tip during the fabrication process. However, one of the current challenges in ion channel measurements is maintaining a single-channel recording. Multiple protein insertions complicate data analysis and destabilize the bilayer. Herein, we combine the promising probe material (Ag/AgCl) with an approach based on current feedback-controlled tip positioning to maintain long-term single-channel recordings for up to 3 h. We develop a hybrid positioning control system, where the channel current is used as feedback to control the vertical movement of the silver tip and, subsequently, control the number of protein channels inserted in the lipid membrane. Our findings reveal that the area of the lipid bilayer decreases with moving the silver tip up (i.e., decreasing the displacement in the z-direction). By reducing the bilayer area around the fine silver tip, we minimize the probability of multiple insertions and remove unwanted proteins. In addition, we characterize the effect of lipid properties such as fluidity on the lipid membrane area. We believe that the use of silver nanoneedles, which enables DC stable channel current, coupled with the developed tip displacement mechanism will offer more opportunities to employ these probes for chemical imaging and mapping different surfaces.

Recent advances in ion-channel probes for nanopore sensing: Insights into the probe architectures.

This review introduces the recent advances in the nanopore sensing platform, ion channel probes (ICPs), with a particular focus on the different probe design (2011-2022). The use of ion channel proteins has emerged in different applications to understand the dynamics of many biological processes and characterize or detect biomolecules. The development of utilizing protein channels in nanopore sensing has led to diverse platforms in which the ion channels, or biological nanopores, can be embedded in a lipid membrane. Ion channel probes, where the ion channels are integrated at the tip of a solid probe, enable higher spatially-resolved detection of small molecules and extend the applications of ion channels to map different surfaces and perform chemical imaging. Different probe materials and designs have been exploited throughout the last decade, which opens the door for multiple probe architecture and applications. We provide more insights into the advances of ICP designs that render them well-suited for further applications.

Silver Nanoneedle Probes Enable Sustained DC Current, Single-Channel Resistive Pulse Nanopore Sensing.

Resistive pulse sensing using ion channel proteins (biological nanopores) has been evolving as a single-molecule approach to detect small biomolecules owing to atomically precise pore size reproducibility, high signal-to-noise ratio, and molecular selectivity. The incorporation of biological nanopores in sensing platforms requires a stable lipid membrane that can be formed by a variety of methods such as the painting method and droplet-based techniques. However, these methods are limited by the fragility of the unsupported bilayer or the need for specific microdevices. Electrode-supported bilayers, in which a metal electrode is used as a support structure, have been recently developed using a fine gold nanoneedle. We previously described the utility of the gold nanoneedle-supported ion channel probe to detect small molecules with high spatial resolution; however, it exhibited a channel current decay over time, which affected the binding frequency of the target molecule to the protein pore as well. Here, we introduce a silver nanoneedle probe to support the lipid bilayer formation and ion channel measurements. The silver nanoneedle mitigates the current decay observed on gold electrodes and produces stable DC channel currents. Our findings propose the formation of a AgCl layer creating a nonpolarizable electrode. The new nanoneedle is successfully applied for single-molecule detection of sulfonated ß-cyclodextrin (S7ßCD) using αHL as a test bed protein. We believe that this new silver nanoneedle platform has great potential given the relative ease of lipid bilayer formation and stable open channel currents.

Development and Validation of Ecofriendly HPLC-MS Method for Quantitative Assay of Amoxicillin, Dicloxacillin, and Their Official Impurity in Pure and Dosage Forms.

Novel, accurate, selective, and rapid high-performance liquid chromatography mass spectrometry method was developed for simultaneous analysis of amoxicillin trihydrate, dicloxacillin sodium, and their official impurity 6-aminopenicillanic acid. The chromatographic separation was carried out by applying the mixture on a C18 column (3.5 µm ps, 100 mm × 4.6 mm id) using acetonitrile:water (65 : 35 by volume) as a mobile phase within only 4 min. The quantitative analysis was executed using single quadrupole mass spectrometer in which electrospray ionization, selected ion monitoring, and negative mode were operated. The retention times were 1.61, 2.54, and 3.50 mins for amoxicillin, 6-aminopenicillanic acid, and dicloxacillin, respectively. The method was validated in linear ranges of 2-28 µg mL-1, 2-35 µg mL-1, and 1-10 µg mL-1 for amoxicillin, dicloxacillin, and 6-aminopenicillanic acid, respectively. The results obtained from the suggested HPLC/MS were statistically compared with those obtained from the reported HPLC method, where no significant difference appeared respecting accuracy and precision. According to the analytical eco-scale assessment method, the proposed method was proved to be greener than the reported one, where the analysis time and the amount of the wasted effluent decreased.

MXene Nanosheets May Induce Toxic Effect on the Early Stage of Embryogenesis.

MXene (Ti3C2Tx), as a novel 2D material, has produced a great interest due to its promising properties in biomedical applications, nevertheless, there is a lack of studies dedicated to investigate the possible toxic effect of MXene in embryos. Herein, we aim to scrutinize the potential toxicity of MXene nanosheets on the early stage of the embryo as well as angiogenesis. Avian embryos at 3 and 5 days of incubation were used as an experimental model in this investigation. Our findings reveal that MXene may produce adverse effect on the early stage of embryogenesis as ∼46% of MXene-exposed embryos died during 1-5 days after exposure. We also found that MXene at tested concentration inhibits angiogenesis of the chorioallantoic membrane of the embryo after 5 days of incubation. More significantly, RT-PCR analysis of seven genes, which are key regulators of cell proliferation, survival, cell death and angiogenesis, revealed that these genes were deregulated in brain, heart and liver tissues from MXene-treated embryos in comparison with their matched controls. Our study clearly suggests that MXene at studied concentration might induce a toxic effect on the early stage of embryogenesis; nevertheless, more investigations are necessary to understand the effect at low concentrations and elucidate its mechanism at the early stage of normal development.

Plasmonic MXene-based nanocomposites exhibiting photothermal therapeutic effects with lower acute toxicity than pure MXene.

Purpose: Here, we fabricated two plasmonic 2D Ti3C2Tx-based nanocomposites (Au/MXene and Au/Fe3O4/MXene) with similarly high anti-cancer photothermal therapy (PTT) capabilities, but with less in vivo toxicity than a pure MXene. Methods: Au/MXene was synthesized by in situ reduction of tetrachloroauric acid using NaBH4 on Ti3C2Tx flakes. For targeted PTT, magnetic Au/Fe3O4/MXene was synthesized via a reaction between freshly prepared magnetite Fe3O4 NPs and MXene solution, followed by in situ integration of gold nanoparticles (AuNPs). Results: Morphological characterization by XRD, SEM, and TEM revealed the successful synthesis of Au/MXene and Au/Fe3O4/MXene. Both new composites exhibited a significant in vitro dose-dependent PTT effect against human breast cancer cells MCF7. Interestingly, in vivo acute toxicity assays using zebrafish embryos indicated that Au/MXene and Au/Fe3O4/MXene had less embryonic mortality (LC50 ≫ 1000 µg/mL) than pure MXene (LC50=257.46 µg/mL). Conclusion: Our new Au/MXene and Au/Fe3O4/MXene nanocomposites could be safer and more suitable than the pure MXene for biomedical applications, especially when targeted PTT is warranted.

Melt Electrospinning Designs for Nanofiber Fabrication for Different Applications.

Nanofibers have been attracting growing attention owing to their outstanding physicochemical and structural properties as well as diverse and intriguing applications. Electrospinning has been known as a simple, flexible, and multipurpose technique for the fabrication of submicro scale fibers. Throughout the last two decades, numerous investigations have focused on the employment of electrospinning techniques to improve the characteristics of fabricated fibers. This review highlights the state of the art of melt electrospinning and clarifies the major categories based on multitemperature control, gas assist, laser melt, coaxial, and needleless designs. In addition, we represent the effect of melt electrospinning process parameters on the properties of produced fibers. Finally, this review summarizes the challenges and obstacles connected to the melt electrospinning technique.

Recent advances in functional nanostructures as cancer photothermal therapy.

Being a non-invasive and relatively safe technique, photothermal therapy has attracted a lot of interest in the cancer treatment field. Recently, nanostructure technology has entered the forefront of cancer therapy owing to its ability to absorb near-infrared radiation as well as efficient light to heat conversion. In this study, key nanostructures for cancer therapy including gold nanoparticles, magnetite iron oxide nanoparticles, organic nanomaterials, and novel two-dimensional nanoagents such as MXenes are discussed. Furthermore, we briefly discuss the characteristics of the nanostructures of these photothermal nanomaterial agents, while focusing on how nanostructures hold potential as cancer therapies. Finally, this review offers promising insight into new cancer therapy approaches, particularly in vivo and in vitro cancer treatments.

Recent Overviews in Functional Polymer Composites for Biomedical Applications.

Composite materials are considered as an essential part of our daily life due to their outstanding properties and diverse applications. Polymer composites are a widespread class of composites, characterized by low cost, facile processing methods, and varied applications ranging from daily-use issues to highly complicated electronics and advanced medical combinations. In this review, we focus on the most important fabrication techniques for bioapplied polymer composites such as electrospinning, melt-extrusion, solution mixing, and latex technology, as well as in situ methods. Additionally, significant and recent advances in biomedical applications are spotlighted, such as tissue engineering (including bone, blood vessels, oral tissues, and skin), dental resin-based composites, and wound dressing.

Partial Least-Squares and Linear Support Vector Regression Chemometric Methods for Simultaneous Determination of Amoxicillin Trihydrate and Dicloxacillin Sodium in the Presence of Their Common Impurity.

Two multivariate chemometric models, namely, partial least-squares regression (PLSR) and linear support vector regression (SVR), are presented for the analysis of amoxicillin trihydrate and dicloxacillin sodium in the presence of their common impurity (6-aminopenicillanic acid) in raw materials and in pharmaceutical dosage form via handling UV spectral data and making a modest comparison between the two models, highlighting the advantages and limitations of each. For optimum analysis, a three-factor, four-level experimental design was established, resulting in a training set of 16 mixtures containing different ratios of interfering species. To validate the prediction ability of the suggested models, an independent test set consisting of eight mixtures was used. The presented results show the ability of the two proposed models to determine the two drugs simultaneously in the presence of small levels of the common impurity with high accuracy and selectivity. The analysis results of the dosage form were statistically compared to a reported HPLC method, with no significant difference regarding accuracy and precision, indicating the ability of the suggested multivariate calibration models to be reliable and suitable for routine analysis of the drug product. Compared to the PLSR model, the SVR model gives more accurate results with a lower prediction error, as well as high generalization ability; however, the PLSR model is easy to handle and fast to optimize.

Development and Validation of HPLC and HPTLC Methods for Determination of Cefoperazone and Its Related Impurities.

Validated sensitive and highly selective methods were developed for the quantitative determination of cefoperazone sodium (CEF) in the presence of its reported impurities; 7-aminocephalosporanic acid (7-ACA) and 5-mercapto-1-methyl-tetrazole (5-MER). Method A is high-performance liquid chromatography (HPLC), where the mixture of CEF and the reported impurities; 7-ACA and 5-MER were separated on a C8 column (5 µm ps, 250 mm × 4.6 i.d.) using methanol:0.05 M KH2PO4 buffer (22.5:77.5 v/v, pH 7.5) as a mobile phase. The three components were detected at 254 nm with a concentration range of 10-90 µg mL(-1) and the mean percentage recovery 99.67% (SD 1.465). Method B is high-performance thin layer chromatography (HPTLC), where the mixture of CEF and the reported impurities were separated on silica gel HPTLC F254 plates using (acetone:methanol:ethyl acetate:2% sodium lauryl sulfate:glacial acetic acid) (3:2:3:0.8:0.2, by volume) as a developing system and scanning at 254 nm over a concentration range of 1-10 µg per band with the mean percentage recovery 99.95% (SD 1.335). The proposed methods were statistically compared with a reported HPLC method with no significant difference regarding accuracy and precision; indicating the ability of the proposed methods to be reliable and suitable for routine analysis of drug product. The proposed HPTLC method proved to be more sensitive, while the HPLC gave more reproducible results besides saving time.

Determination of Cefoperazone Sodium in Presence of Related Impurities by Linear Support Vector Regression and Partial Least Squares Chemometric Models.

A comparison between partial least squares regression and support vector regression chemometric models is introduced in this study. The two models are implemented to analyze cefoperazone sodium in presence of its reported impurities, 7-aminocephalosporanic acid and 5-mercapto-1-methyl-tetrazole, in pure powders and in pharmaceutical formulations through processing UV spectroscopic data. For best results, a 3-factor 4-level experimental design was used, resulting in a training set of 16 mixtures containing different ratios of interfering moieties. For method validation, an independent test set consisting of 9 mixtures was used to test predictive ability of established models. The introduced results show the capability of the two proposed models to analyze cefoperazone in presence of its impurities 7-aminocephalosporanic acid and 5-mercapto-1-methyl-tetrazole with high trueness and selectivity (101.87 ± 0.708 and 101.43 ± 0.536 for PLSR and linear SVR, resp.). Analysis results of drug products were statistically compared to a reported HPLC method showing no significant difference in trueness and precision, indicating the capability of the suggested multivariate calibration models to be reliable and adequate for routine quality control analysis of drug product. SVR offers more accurate results with lower prediction error compared to PLSR model; however, PLSR is easy to handle and fast to optimize.