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%).

Addressing the Needs of the Rapidly Aging Society through the Development of Multifunctional Bioactive Coatings for Orthopedic Applications.

The unprecedented aging of the world's population will boost the need for orthopedic implants and expose their current limitations to a greater extent due to the medical complexity of elderly patients and longer indwelling times of the implanted materials. Biocompatible metals with multifunctional bioactive coatings promise to provide the means for the controlled and tailorable release of different medications for patient-specific treatment while prolonging the material's lifespan and thus improving the surgical outcome. The objective of this work is to provide a review of several groups of biocompatible materials that might be utilized as constituents for the development of multifunctional bioactive coatings on metal materials with a focus on antimicrobial, pain-relieving, and anticoagulant properties. Moreover, the review presents a summary of medications used in clinical settings, the disadvantages of the commercially available products, and insight into the latest development strategies. For a more successful translation of such research into clinical practice, extensive knowledge of the chemical interactions between the components and a detailed understanding of the properties and mechanisms of biological matter are required. Moreover, the cost-efficiency of the surface treatment should be considered in the development process.

Gynaecological cancers and their cell lines.

Cell lines are widely used for various research purposes including cancer and drug research. Recently, there have been studies that pointed to discrepancies in the literature and usage of cell lines. That is why we have prepared a comprehensive overview of the most common gynaecological cancer cell lines, their literature, a list of currently available cell lines, and new findings compared with the original studies. A literature review was conducted via MEDLINE, PubMed and ScienceDirect for reviews in the last 5 years to identify research and other studies related to gynaecological cancer cell lines. We present an overview of the current literature with reference to the original studies and pointed to certain inconsistencies in the literature. The adherence to culturing rulesets and the international guidelines helps in minimizing replication failure between institutions. Evidence from the latest research suggests that despite certain drawbacks, variations of cancer cell lines can also be useful in regard to a more diverse genomic landscape.

Progressive use of multispectral imaging flow cytometry in various research areas.

Multi-spectral imaging flow cytometry (MIFC) has become one of the most powerful technologies for investigating general analytics, molecular and cell biology, biotechnology, medicine, and related fields. It combines the capabilities of the morphometric and photometric analysis of single cells and micrometer-sized particles in flux with regard to thousands of events. It has become the tool of choice for a wide range of research and clinical applications. By combining the features of flow cytometry and fluorescence microscopy, it offers researchers the ability to couple the spatial resolution of multicolour images of cells and organelles with the simultaneous analysis of a large number of events in a single system. This provides the opportunity to visually confirm findings and collect novel data that would otherwise be more difficult to obtain. This has led many researchers to design innovative assays to gain new insight into important research questions. To date, it has been successfully used to study cell morphology, surface and nuclear protein co-localization, protein-protein interactions, cell signaling, cell cycle, cell death, and cytotoxicity, intracellular calcium, drug uptake, pathogen internalization, and other applications. Herein we describe some of the recent advances in the field of multiparametric imaging flow cytometry methods in various research areas.

Effects of the reservoir bag disconnection on inspired gases during general anesthesia: a simulator-based study.

BACKGROUND: Fresh gas decoupling is a feature of the modern anesthesia workstation, where the fresh gas flow (FGF) is diverted into the reservoir bag and is not added to the delivered tidal volume, which thus remains constant. The present study aimed to investigate the entraining of the atmospheric air into the anesthesia breathing circuit in case the reservoir bag was disconnected. METHODS: We conducted a simulator-based study, where the METI HPS simulator was connected to the anesthesia workstation. The effect of the disconnected reservoir bag was evaluated using oxygen (O2) and air or oxygen and nitrous oxide (N2O) as a carrier gas at different FGF rates. We disconnected the reservoir bag for 10 min during the maintenance phase. We recorded values for inspiratory O2, N2O, and sevoflurane. The time constant of the exponential process was estimated during reservoir bag disconnection. RESULTS: The difference of O2, N2O and sevoflurane concentrations, before, during, and after reservoir bag disconnection was statistically significant at 0.5, 1, and 2 L/min of FGF (p < 0.001). The largest decrease of the inspired O2 concentrations (FIO2) was detected in the case of oxygen and air as the carrier gas and an FGF of 1 L/min, when oxygen decreased from median [25th-75th percentile] 55.00% [54.00-56.00] to median 39.50% [38.00-42.50] (p < 0.001). The time constant for FIO2 during reservoir bag disconnection in oxygen and air as the carrier gas, were median 2.5, 2.5, and 1.5 min in FGF of 0.5, 1.0, and 2 L/min respectively. CONCLUSIONS: During the disconnection of the anesthesia reservoir bag, the process of pharmacokinetics takes place faster compared to the wash-in and wash-out pharmacokinetic properties in the circle breathing system. The time constant was affected by the FGF rate, as well as the gradient of anesthetic gases between the anesthesia circle system and atmospheric air.

Isolation, characterisation and phagocytic function of human macrophages from human peripheral blood.

Macrophages are among the most important cells of the immune system. Among other functions, they take part in almost all defense actions against foreign bodies and bacteria, being particularly important in infections, wound healing, and foreign body reactions. Considering their importance for the health of the human body, as well as their important role in several diseases, the in vitro studies based on these cells, are a crucial research field. Taking all mentioned into account, this study describes a simple isolation method of human macrophages (MFUM-HMP-001 and MFUM-HMP-002 cell lines) from peripheral blood. For this purpose, the morphology, the viability, and the phagocytotic activity of the isolated cells were tested. The Immunostaining of MFUM-HMP-001 and MFUM-HMP-002 cells confirmed the macrophage cell markers CD68, CD80, and CD163/M130. The phagocytotic activity was marked in both MFUM-HMP-001 and MFUM-HMP-002 cells, as was the phagocytosis of the pHrodo green Escherichia coli bioparticles conjugates, which was enhanced with the addition of lipopolysaccharide. The cells were stable and exhibited good growth. According to our results, both cell lines are useful for the development of novel macrophage cell-based in vitro models.

Renal proximal tubular epithelial cells: review of isolation, characterization, and culturing techniques.

The kidney is a complex organ, comprised primarily of glomerular, tubular, mesangial, and endothelial cells, and podocytes. The fact that renal cells are terminally differentiated at 34 weeks of gestation is the main obstacle in regeneration and treatment of acute kidney injury or chronic kidney disease. Furthermore, the number of chronic kidney disease patients is ever increasing and with it the medical community should aim to improve existing and develop new methods of renal replacement therapy. On the other hand, as polypharmacy is on the rise, thought should be given into developing new ways of testing drug safety. A possible way to tackle these issues is with isolation and culture of renal cells. Several protocols are currently described to isolate the desired cells, of which the most isolated are the proximal tubular epithelial cells. They play a major role in water homeostasis, acid-base control, reabsorption of compounds, and secretion of xenobiotics and endogenous metabolites. When exposed to ischemic, toxic, septic, or obstructive conditions their death results in what we clinically perceive as acute kidney injury. Additionally, due to renal cells' limited regenerative potential, the profibrotic environment inevitably leads to chronic kidney disease. In this review we will focus on human proximal tubular epithelial cells. We will cover human kidney culture models, cell sources, isolation, culture, immortalization, and characterization subdivided into morphological, phenotypical, and functional characterization.

Endometrial cancer and its cell lines.

Endometrial cancer is one of the most common gynaecological malignancies worldwide. One type of research in this field is the growing of cell lines (CLs) and cultures, which can be used to explore the biological mechanisms of cancer. The purpose of this review is to offer an overview of the current literature and highlight the importance of correct CL studies. We carried out a literature analysis of more than 60 articles from the Pubmed, Medline databases that were almost exclusively published in indexed journals in the last 10 years as well as the primary originating scientific studies of specific CLs. We then summarized the newest findings and recommendations. Cell lines are becoming widely used as in vitro tumour models. Recent work has shown inconsistencies in nomenclature and culturing of CLs. Their genomic evolution leads to a high degree of variation across CL strains therefore it is of the utmost importance to recognize the variability within laboratory cancer models. Laboratories must adapt, incorporate additional characterisation techniques and view this situation as an opportunity to improve the reproducibility of pre-clinical cancer research. The authors offer a comprehensive literature review about endometrial cancer CLs, a review of the current literature and advice on culturing CLs.

Nano- and Micropatterned Polycaprolactone Cellulose Composite Surfaces with Tunable Protein Adsorption, Fibrin Clot Formation, and Endothelial Cellular Response.

This work describes the interaction of the human blood plasma proteins albumin, fibrinogen, and γ-globulins with micro- and nanopatterned polymer interfaces. Protein adsorption studies were correlated with the fibrin clotting time of human blood plasma and with the growth of primary human pulmonary artery endothelial cells (hECs) on these patterns. It was observed that blends of polycaprolactone (PCL) and trimethylsilyl-protected cellulose form various thin-film patterns during spin coating, depending on the mass ratio of the polymers in the spinning solutions. Vapor-phase acid-catalyzed deprotection preserves these patterns but yields interfaces that are composed of hydrophilic cellulose domains enclosed by hydrophobic PCL. The blood plasma proteins are repelled by the cellulose domains, allowing for a suggested selective protein deposition on the PCL domains. An inverse proportional correlation is observed between the amount of cellulose present in the films and the mass of irreversibly adsorbed proteins. This results in significantly increased fibrin clotting times and lower masses of deposited clots on cellulose-containing films as revealed by quartz crystal microbalance with dissipation measurements. Cell viability of hECs grown on these surfaces was directly correlated with higher protein adsorption and faster clot formation. The results show that presented patterned polymer composite surfaces allow for a controllable blood plasma protein coagulation and a significant biological response from hECs. It is proposed that this knowledge can be utilized in regenerative medicine, cell cultures, and artificial vascular grafts by a careful choice of polymers and patterns.

Genetic biases related to chronic venous ulceration.

Chronic wounds represent a major socioeconomic problem. Chronic venous ulceration is one of the least well-understood types of chronic wounds. A chronic venous ulcer arises as a result of chronic venous insufficiency (CVI), which affects approximately 10-35% of people in the developed world, yet not all people with CVI develop ulceration. The question of why some patients with CVI develop chronic ulceration and others do not, still remains unanswered. Risk factors for the development of chronic ulceration are poorly understood and include age, residual iliofemoral vein obstruction, residual deep incompetence, persistent venous hypertension, obesity and genetics. The genetic aspects of CVI have only been vaguely evaluated. This paper reports on a literature review of the variation in genetic polymorphisms and gene expression associated with the development of a chronic venous ulceration.

Matrix Tablets for Controlled Release of Drugs Incorporated Using Capillary Absorption.

Cost and time effectiveness make direct tableting still the favored method for tablet production. Among its most noticeable limitations in application is the non-uniformity (and/or inhomogeneities) in the contents of the resulting tablets, possibly leading to inconsistencies in required tablet properties. The efficiency of direct tableting is mostly affected by surface properties of the components to be tableted, which govern the final tablet mechanical and chemical properties and can influence the liquid capillary rise that the tablets exhibit after ingestion. By using capillary rise as a driving force, we developed a simple, yet powerful procedure for filling blank tablets with a repeatable drug amount. Blank tablets were prepared by direct compression of the excipient and filled with an organic solution of hydrochlorothiazide. Tablets were characterized regarding their structure and morphology, while their applicability was monitored using in vitro drug release studies. By utilizing the mentioned filling of blank tablets, we were able to incorporate the desired dose of the drug inside while maintaining the tablets initial mechanical properties. Moreover, most of the drug was incorporated in the tablet pores and the rest was homogeneously distributed over the tablet surface in the form of small particles, by which we also eliminated content non-uniformity (homogenous drug distribution through the tablet). To sum up, we not only developed a cheap, simple, and reproducible variation of direct tableting, but were also able to eliminate some of its biggest disadvantages (e.g., segregation of components, leading to inhomogeneities in contents, and incompatibility between different base ingredients due to their different surface properties). All mentioned make the proposed approach highly interesting for future use, especially in potential therapy individualization.

Systematic Evaluation of a Diclofenac-Loaded Carboxymethyl Cellulose-Based Wound Dressing and Its Release Performance with Changing pH and Temperature.

Development of drug-loaded wound dressings is often performed without systematic consideration of the changing wound environment that can influence such materials' performance. Among the crucial changes are the wound pH and temperature, which have an immense effect on the drug release. Detailed release studies based on the consideration of these changing properties provide an important aspect of the in vitro performance testing of novel wound dressing materials. A sodium carboxymethyl cellulose-based wound dressing, with the incorporated non-steroidal anti-inflammatory drug diclofenac, was developed and characterised in regard to its physico-chemical, structural and morphological properties. Further, the influence of pH and temperature were studied on the drug release. Finally, the biocompatibility of the wound dressing towards human skin cells was tested. Incorporation of diclofenac did not alter important properties (water retention value, air permeability) of the host material. Changes in the pH and temperature were shown to influence the release performance and have to be accounted for in the evaluation of such dressings. Furthermore, the knowledge about the potential changes of these parameters in the wound bed could be used potentially to predict, and potentially even to control the drug release from the developed wound dressing. The prepared wound dressing was also proven biocompatible towards human skin cells, making it interesting for potential future use in the clinics.

Reusability of SPE and Sb-modified SPE Sensors for Trace Pb(II) Determination.

In this work, unmodified screen-printed electrode (bare SPE) and Sb-film modified SPE (SbFSPE) sensors were employed for the analysis of trace amounts of Pb(II) in non-deaerated water solutions. The modified electrode was performed in situ in 0.5 mg/L Sb(III) and 0.01 M HCl. The methodology was validated for an accumulation potential of ⁻1.1 V vs. Ag/AgCl and an accumulation time of 60 s. A comparative analysis of bare SPE and SbFSPE showed that the detection and quantification limits decrease for the bare SPE. The method with the bare SPE showed a linear response in the 69.8⁻368.4 µg/L concentration range, whereas linearity for the SbFSPE was in the 24.0⁻319.1 µg/L concentration range. This work also reports the reason why the multiple standard addition method instead of a linear calibration curve for Pb(II) analysis should be employed. Furthermore, the analytical method employing SbFSPE was found to be more accurate and precise compared to the use of bare SPE when sensors were employed for the first time, however this performance changed significantly when these sensors were reused in the same manner. Furthermore, electrochemical impedance spectroscopy was used for the first time to analyse the electrochemical response of sensors after being used for multiple successive analyses. Surface characterisation before and after multiple successive uses of bare SPE and SbFSPE sensors, with atomic force microscopy and field emission scanning electron microscopy, showed sensor degradation. The interference effect of Cd(II), Zn(II), As(III), Fe(II), Na(I), K(I), Ca(II), Mg(II), NO3⁻, Bi(III), Cu(II), Sn(II), and Hg(II) on the Pb(II) stripping signal was also studied. Finally, the application of SbFSPE was tested on a real water sample (from a local river), which showed high precision (RSD = 8.1%, n = 5) and accurate results.

Multilayered Polysaccharide Nanofilms for Controlled Delivery of Pentoxifylline and Possible Treatment of Chronic Venous Ulceration.

Local drug delivery systems made from nontoxic polysaccharide nanofilms have an enormous potential in wound care. A detailed understanding of the structural, surface, physicochemical, and cytotoxic properties of such systems is crucial to design clinically efficacious materials. Herein, we fabricated polysaccharide-based nanofilms onto either a 2D model (SiO2 and Au sensors) or on nonwoven alginate 3D substrates using an alternating assembly of N,N,N-trimethylchitosan (TMC) and alginic acid (ALG) by a spin-assisted layer-by-layer (LbL) technique. These TMC/ALG multilayered nanofilms are used for a uniform encapsulation and controlled release of pentoxifylline (PTX), a potent anti-inflammatory drug for treatment of the chronic venous ulceration. We show a tailorable film growth and mass, morphology, as well as surface properties (charge, hydrophilicity, porosity) of the assembled nanofilms through control of the coating during the spin-assisted assembly. The uniform distribution of the encapsulated PTX in the TMC/ALG nanofilms is preserved even with when the amount of the incorporated PTX increases. The PTX release mechanism from the model and real systems is studied in detail and is very comparable for both systems. Finally, different cell-based assays illustrated the potential of the TMC/ALG multilayer system in wound care (e.g., treatment chronic venous ulceration) applications, including a decrease of TNF-α secretion, a common indicator of inflammation.

Utilization of optical polarization microscopy in the study of sorption characteristics of wound dressing host materials.

Polarization microscopy was used for evaluation of kinetics of diclofenac sorption in three different wound dressing materials. The sorption kinetics can be evaluated by radii change and intensity of the light traveling through the fiber. The most frequently used host materials for drugs in wound dressings are alginate, polyesters such as polyethylene terephthalate, and viscose. We studied sorption of diclofenac as an example drug. Effective, but rather simple in vitro simulation of diclofenac sorption gives insight into the applicability of the mentioned materials for development of wound healing materials.

Editorial on the Special Issue Entitled "Recent Advances in Aerogels".

Aerogels are unique solid materials that consist mainly of air and have an extremely low density, large open pores, and a large internal surface area [...].

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.

Review of Potential Drug-Eluting Contact Lens Technologies.

The field of ophthalmology is expanding exponentially, both in terms of diagnostic and therapeutic capabilities, as well as the worldwide increasing incidence of eye-related diseases. Due to an ageing population and climate change, the number of ophthalmic patients will continue to increase, overwhelming healthcare systems and likely leading to under-treatment of chronic eye diseases. Since drops are the mainstay of therapy, clinicians have long emphasised the unmet need for ocular drug delivery. Alternative methods, i.e., with better compliance, stability and longevity of drug delivery, would be preferred. Several approaches and materials are being studied and used to overcome these drawbacks. We believe that drug-loaded contact lenses are among the most promising and are a real step toward dropless ocular therapy, potentially leading to a transformation in clinical ophthalmic practice. In this review, we outline the current role of contact lenses in ocular drug delivery, focusing on materials, drug binding and preparation, concluding with a look at future developments.

Cell Electrospinning: A Mini-Review of the Critical Processing Parameters and Its Use in Biomedical Applications.

Functional tissue engineering is a widely studied area of research with increasing importance in regenerative medicine, as well as in the development of in vitro models used for drug discovery and mimicking diseased tissues, among other applications. Electrospinning (ES) is one of the most widely used methods in these fields. It has attracted considerable interest because it can produce materials resembling the extracellular matrix of native tissues. The micro/nanofibers produced by this method provide a cell-friendly environment that promotes cellular activities. Cell electrospinning (C-ES) is based on the fundamental ES process and enables the encapsulation of viable cells in a micro/nanofibrous mesh. In this review, the process of C-ES and the materials used in this process are discussed. This work also discusses the applications of C-ES in tissue engineering, focusing on recent advances in this field.