The development of an electropolymerized, molecularly imprinted polymer (MIP) sensor for insulin determination using single-drop analysis.

An electrochemical sensor for the detection of insulin in a single drop (50 µL) was developed based on the concept of molecularly imprinted polymers (MIP). The synthetic MIP receptors were assembled on a screen-printed carbon electrode (SPCE) by the electropolymerization of pyrrole (Py) in the presence of insulin (the protein template) using cyclic voltammetry. After electropolymerization, insulin was removed from the formed polypyrrole (Ppy) matrix to create imprinting cavities for the subsequent analysis of the insulin analyte in test samples. The surface characterization, before and after each electrosynthesis step of the MIP sensors, was performed using atomic force microscopy, scanning electron microscopy, and energy-dispersive X-ray spectroscopy. The performance of the developed MIP-SPCE sensor was evaluated using a single drop of solution containing K3Fe(CN)6 and the square-wave voltammetry technique. The MIP-SPCE showed a linear concentration range of 20.0-70.0 pM (R2 = 0.9991), a limit of detection of 1.9 pM, and a limit of quantification of 6.2 pM. The rapid response time to the protein target and the portability of the developed sensor, which is considered a disposable MIP-based system, make this MIP-SPCE sensor a promising candidate for point-of-care applications. In addition, the MIP-SPCE sensor was successfully used to detect insulin in a pharmaceutical sample. The sensor was deemed to be accurate (the average recovery was 108.46%) and precise (the relative standard deviation was 7.23%).

Nanostructured Bismuth Film Electrode for Detection of Progesterone.

Progesterone is an important hormone responsible, among others, for maintaining pregnancy via inhibition of uterus muscles activity; thus, following its concentration levels in pregnant women is of immense importance in the endeavor to prevent premature birth. In this work, the nanostructured bismuth film electrode (nsBiFE) was studied for detection of progesterone in neutral medium. Due to the ability to accumulate progesterone at the nsBiFE, the adsorptive cathodic stripping voltammetry was beneficially exploited. The nsBiFE was prepared on the surface of a glassy carbon supporting electrode and several parameters influencing the detection of progesterone were investigated. The nsBiFE exhibited superior electroanalytical characteristics in comparison to other bismuth-based electrodes and unmodified glassy carbon electrode together with satisfactory response toward low concentrations of progesterone, which are consistent with clinically significant levels.

Novel 3D printed polysaccharide-based materials with prebiotic activity for potential treatment of diaper rash.

Diaper rash, mainly occurring as erythema and itching in the diaper area, causes considerable distress to infants and toddlers. Increasing evidence suggests that an unequal distribution of microorganisms on the skin contributes to the development of diaper dermatitis. Probiotic bacteria, like Staphylococcus epidermidis, are crucial for maintaining a healthy balance in the skin's microbiome, among others, through their fermentative metabolites, such as short-chain fatty acids. Using a defined prebiotic as a carbon source (e.g., as part of the diaper formulation) can selectively trigger the fermentation of probiotic bacteria. A proper material choice can reduce diaper rash incidence by diminishing the skin exposure to wetness and faeces. Using 3D printing, we fabricated carbon-rich materials for the top sheet layer of baby diapers that enhance the probiotic activity of S. epidermidis. The developed materials' printability, chemical composition, swelling ability, and degradation rate were analysed. In addition, microbiological tests evaluated their potential as a source of in situ short-chain fatty acid production. Finally, biocompatibility testing with skin cells evaluated their safety for potential use as part of diapers. The results demonstrate a cost-effective approach for producing novel materials that can tailor the ecological balance of the skin microflora and help treat diaper rash.

Multilayer Methacrylate-Based Wound Dressing as a Therapeutic Tool for Targeted Pain Relief.

This study presents an innovative wound dressing system that offers a highly effective therapeutic solution for treating painful wounds. By incorporating the widely used non-steroidal anti-inflammatory drug diclofenac, we have created an active wound dressing that can provide targeted pain relief with ease. The drug was embedded within a biocompatible matrix composed of polyhydroxyethyl methacrylate and polyhydroxypropyl methacrylate. The multilayer structure of the dressing, which allows for sustained drug release and an exact application, was achieved through the layer-by-layer coating technique and the inclusion of superparamagnetic iron platinum nanoparticles. The multilayered dressings' physicochemical, structural, and morphological properties were characterised using various methods. The synergistic effect of the incorporated drug molecules and superparamagnetic nanoparticles on the surface roughness and release kinetics resulted in controlled drug release. In addition, the proposed multilayer wound dressings were found to be biocompatible with human skin fibroblasts. Our findings suggest that the developed wound dressing system can contribute to tailored therapeutic strategies for local pain relief.

Astrocytes and human artificial blood-brain barrier models.

The blood-brain barrier (BBB) functions as a highly selective border of endothelial cells, protecting the central nervous system from potentially harmful substances by selectively controlling the entry of cells and molecules, including components of the immune system. To study the BBB properties, find suitable therapies, and identify new drug targets, there is a need to develop representative in vitro BBB models. In this article, we describe the astrocyte roles in the BBB functioning and human in vitro BBB models.

Artificial Biomimetic Electrochemical Assemblies.

Rapid, selective, and cost-effective detection and determination of clinically relevant biomolecule analytes for a better understanding of biological and physiological functions are becoming increasingly prominent. In this regard, biosensors represent a powerful tool to meet these requirements. Recent decades have seen biosensors gaining popularity due to their ability to design sensor platforms that are selective to determine target analytes. Naturally generated receptor units have a high affinity for their targets, which provides the selectivity of a device. However, such receptors are subject to instability under harsh environmental conditions and have consequently low durability. By applying principles of supramolecular chemistry, molecularly imprinted polymers (MIPs) can successfully replace natural receptors to circumvent these shortcomings. This review summarizes the recent achievements and analytical applications of electrosynthesized MIPs, in particular, for the detection of protein-based biomarkers. The scope of this review also includes the background behind electrochemical readouts and the origin of the gate effect in MIP-based biosensors.

Novel Methacrylate-Based Multilayer Nanofilms with Incorporated FePt-Based Nanoparticles and the Anticancer Drug 5-Fluorouracil for Skin Cancer Treatment.

Despite medical advances, skin-associated disorders continue to pose a unique challenge to physicians worldwide. Skin cancer is one of the most common forms of cancer, with more than one million new cases reported each year. Currently, surgical excision is its primary treatment; however, this can be impractical or even contradictory in certain situations. An interesting potential alternative could lie in topical treatment solutions. The goal of our study was to develop novel multilayer nanofilms consisting of a combination of polyhydroxyethyl methacrylate (PHEMA), polyhydroxypropyl methacrylate (PHPMA), sodium deoxycholate (NaDOC) with incorporated superparamagnetic iron-platinum nanoparticles (FePt NPs), and the potent anticancer drug (5-fluorouracil), for theranostic skin cancer treatment. All multilayer systems were prepared by spin-coating and characterised by atomic force microscopy, infrared spectroscopy, and contact angle measurement. The magnetic properties of the incorporated FePt NPs were evaluated using magnetisation measurement, while their size was determined using transmission electron microscopy (TEM). Drug release performance was tested in vitro, and formulation safety was evaluated on human-skin-derived fibroblasts. Finally, the efficacy for skin cancer treatment was tested on our own basal-cell carcinoma cell line.

Hybrid 3D Printing of Advanced Hydrogel-Based Wound Dressings with Tailorable Properties.

Despite the extensive utilization of polysaccharide hydrogels in regenerative medicine, current fabrication methods fail to produce mechanically stable scaffolds using only hydrogels. The recently developed hybrid extrusion-based bioprinting process promises to resolve these current issues by facilitating the simultaneous printing of stiff thermoplastic polymers and softer hydrogels at different temperatures. Using layer-by-layer deposition, mechanically advantageous scaffolds can be produced by integrating the softer hydrogel matrix into a stiffer synthetic framework. This work demonstrates the fabrication of hybrid hydrogel-thermoplastic polymer scaffolds with tunable structural and chemical properties for applications in tissue engineering and regenerative medicine. Through an alternating deposition of polycaprolactone and alginate/carboxymethylcellulose gel strands, scaffolds with the desired architecture (e.g., filament thickness, pore size, macro-/microporosity), and rheological characteristics (e.g., swelling capacity, degradation rate, and wettability) were prepared. The hybrid fabrication approach allows the fine-tuning of wettability (approx. 50-75°), swelling (approx. 0-20× increased mass), degradability (approx. 2-30+ days), and mechanical strength (approx. 0.2-11 MPa) in the range between pure hydrogels and pure thermoplastic polymers, while providing a gradient of surface properties and good biocompatibility. The controlled degradability and permeability of the hydrogel component may also enable controlled drug delivery. Our work shows that the novel hybrid hydrogel-thermoplastic scaffolds with adjustable characteristics have immense potential for tissue engineering and can serve as templates for developing novel wound dressings.

Polysaccharide-Based Bioink Formulation for 3D Bioprinting of an In Vitro Model of the Human Dermis.

Limitations in wound management have prompted scientists to introduce bioprinting techniques for creating constructs that can address clinical problems. The bioprinting approach is renowned for its ability to spatially control the three-dimensional (3D) placement of cells, molecules, and biomaterials. These features provide new possibilities to enhance homology to native skin and improve functional outcomes. However, for the clinical value, the development of hydrogel bioink with refined printability and bioactive properties is needed. In this study, we combined the outstanding viscoelastic behavior of nanofibrillated cellulose (NFC) with the fast cross-linking ability of alginate (ALG), carboxymethyl cellulose (CMC), and encapsulated human-derived skin fibroblasts (hSF) to create a bioink for the 3D bioprinting of a dermis layer. The shear thinning behavior of hSF-laden bioink enables construction of 3D scaffolds with high cell density and homogeneous cell distribution. The obtained results demonstrated that hSF-laden bioink supports cellular activity of hSF (up to 29 days) while offering proper printability in a biologically relevant 3D environment, making it a promising tool for skin tissue engineering and drug testing applications.

Core/shell Printing Scaffolds For Tissue Engineering Of Tubular Structures.

Three-dimensional (3D) printing of core/shell filaments allows direct fabrication of channel structures with a stable shell that is cross-linked at the interface with a liquid core. The latter is removed post-printing, leaving behind a hollow tube. Integrating an additive manufacturing technique (like the one described here with tailor-made [bio]inks, which structurally and biochemically mimic the native extracellular matrix [ECM]) is an important step towards advanced tissue engineering. However, precise fabrication of well-defined structures requires tailored fabrication strategies optimized for the material in use. Therefore, it is sensible to begin with a set-up that is customizable, simple-to-use, and compatible with a broad spectrum of materials and applications. This work presents an easy-to-manufacture core/shell nozzle with luer-compatibility to explore core/shell printing of woodpile structures, tested with a well-defined, alginate-based scaffold material formulation.

Molecular evidence and high genetic diversity of shrew-borne Seewis virus in Slovenia.

Seewis virus, the shrew-borne hantavirus from Sorex araneus, has been molecularly detected in reservoir hosts in many different central European countries and Russia. Slovenia is a known endemic country for rodent-borne hantaviruses, therefore the aim of the study was to investigate the presence of shrew-borne hantaviruses in insectivores. Viral L, S and M segment have been recovered only from tissue samples of 7 S. araneus, despite several shrew species were tested. Phylogenetic analysis showed high genetic diversity of SWSV in Slovenia, ranging from 3 to 19.4% for different viral segments. The most divergent were M segment sequences, with 19.4% nucleotide divergence among Slovenian strains. Above that, different SWSV strains from Slovenia do not group into separate geographic clusters. While three separate genetic clades were determined, two of them were simultaneously present in one location at the same time.