Novel multifunctional tacrine-donepezil hybrids against Alzheimer's disease: Design synthesis and bioactivity studies.

A series of tacrine-donepezil hybrids were synthesized as potential multifunctional anti-Alzheimer's disease (AD) compounds. For this purpose, tacrine and the benzylpiperidine moiety of donepezil were fused with a hydrazone group to achieve a small library of tacrine-donepezil hybrids. In agreement with the design, all compounds showed inhibitory activity toward both acetylcholinesterase (AChE) and butyrylcholinesterase (BChE) with IC50 values in the low micromolar range. Kinetic studies on the most potent cholinesterase (ChE) inhibitors within the series showed a mixed-type inhibition mechanism on both enzymes. Also, the docking studies indicated that the compounds inhibit ChEs by dual binding site (DBS) interactions. Notably, tacrine-donepezil hybrids also exhibited significant neuroprotection against H2O2-induced cell death in a differentiated human neuroblastoma (SH-SY5Y) cell line at concentrations close to their IC50 values on ChEs and showed high to medium blood-brain barrier (BBB) permeability on human cerebral microvascular endothelial cells (HBEC-5i). Besides, the compounds do not cause remarkable toxicity in a human hepatocellular carcinoma cell line (HepG2) and SH-SY5Y cells. Additionally, the compounds were predicted to also have good bioavailability. Among the tested compounds, H4, H16, H17, and H24 stand out with their biological profile. Taken together, the proposed novel tacrine-donepezil scaffold represents a promising starting point for the development of novel anti-ChE multifunctional agents against AD.

Magnetic Hydrogel Beads as a Reusable Adsorbent for Highly Efficient and Rapid Removal of Aluminum: Characterization, Response Surface Methodology Optimization, and Evaluation of Isotherms, Kinetics, and Thermodynamic Studies.

Biopolymers such as alginate and gelatin have attracted much attention because of their exceptional adsorption properties and biocompatibility. The magnetic hydrogel beads produced and used in this study had a core structure composed of magnetite nanoparticles and gelatin and a shell structure composed of alginate. The combination of the metal-ion binding ability of alginate and the mechanical strength of gelatin in magnetic hydrogel beads presents a new approach for the removal of metal from water sources. The beads were designed for aluminum removal and fully characterized using various methods, including Fourier transform infrared spectroscopy, X-ray photoelectron spectroscopy, scanning electron microscopy-energy-dispersive X-ray spectroscopy, vibrating sample magnetometry, microcomputed tomography, and dynamic mechanical analysis. Statistical experimental designs were employed to optimize the parameters of the adsorption and recovery processes. Plackett-Burman Design, Box-Behnken Design, and Central Composite Design were used for identifying the significant factors and optimizing the parameters of the adsorption and recovery processes, respectively. The optimum parameters determined for adsorption are as follows: pH: 4, contact time: 30 min, adsorbent amount: 600 mg; recovery time: reagent 1 M HNO3; and contact time: 40 min. The adsorption process was described by using the Langmuir isotherm model. It reveals a homogeneous bead surface and monolayer adsorption with an adsorption capacity of 5.25 mg g-1. Limit of detection and limit of quantification values were calculated as 4.3 and 14 µg L-1, respectively. The adsorption process was described by a pseudo-second-order kinetic model, which assumes that chemisorption is the rate-controlling mechanism. Thermodynamic studies indicate that adsorption is spontaneous and endothermic. The adsorbent was reusable for 10 successive adsorption-desorption cycles with a quantitative adsorption of 98.2% ± 0.3% and a recovery of 99.4% ± 2.6%. The minimum adsorbent dose was determined as 30 g L-1 to achieve quantitative adsorption of aluminum. The effects of the inorganic ions were also investigated. The proposed method was applied to tap water and carboy water samples, and the results indicate that magnetic hydrogel beads can be an effective and reusable bioadsorbent for the detection and removal of aluminum in water samples. The recovery values obtained by using the developed method were quantitative and consistent with the results obtained from the inductively coupled plasma optical emission spectrometer.

Grafting of Cyclodextrin to Theranostic Nanoparticles Improves Blood-Brain Barrier Model Crossing.

Core-shell superparamagnetic iron oxide nanoparticles hold great promise as a theranostic platform in biological systems. Herein, we report the biological effect of multifunctional cyclodextrin-appended SPIONs (CySPION) in mutant Npc1-deficient CHO cells compared to their wild type counterparts. CySPIONs show negligible cytotoxicity while they are strongly endocytosed and localized in the lysosomal compartment. Through their bespoke pH-sensitive chemistry, these nanoparticles release appended monomeric cyclodextrins to mobilize over-accumulated cholesterol and eject it outside the cells. CySPIONs show a high rate of transport across blood-brain barrier models, indicating their promise as a therapeutic approach for cholesterol-impaired diseases affecting the brain.

Role of Intermediate Filaments in Blood-Brain Barrier in Health and Disease.

The blood-brain barrier (BBB) is a highly selective cellular monolayer unique to the microvasculature of the central nervous system (CNS), and it mediates the communication of the CNS with the rest of the body by regulating the passage of molecules into the CNS microenvironment. Limitation of passage of substances through the BBB is mainly due to tight junctions (TJ) and adherens junctions (AJ) between brain microvascular endothelial cells. The importance of actin filaments and microtubules in establishing and maintaining TJs and AJs has been indicated; however, recent studies have shown that intermediate filaments are also important in the formation and function of cell-cell junctions. The most common intermediate filament protein in endothelial cells is vimentin. Vimentin plays a role in blood-brain barrier permeability in both cell-cell and cell-matrix interactions by affecting the actin and microtubule reorganization and by binding directly to VE-cadherin or integrin proteins. The BBB permeability increases due to the formation of stress fibers and the disruption of VE-cadherin interactions between two neighboring cells in various diseases, disrupting the fiber network of intermediate filament vimentin in different ways. Intermediate filaments may be long ignored key targets in regulation of BBB permeability in health and disease.

In Vitro Human Blood-Brain Barrier Model for Drug Permeability Testing.

Blood-brain barrier (BBB), although very important for protection of brain from major neurotoxins, negatively affects the treatment of central nervous system diseases by limiting the passage of neuropharmaceuticals from blood to the brain. Thus, researchers have to investigate the passage of the produced drug molecules through the BBB before they are introduced to the market. Although these experiments have been traditionally performed on experimental animals, drug permeability tests are now carried out mostly by in vitro BBB models due to ethical problems, differences between species, and expensive and troublesome in vivo test procedures. In this method, we explain how to model and characterize a realistic in vitro BBB model using human derived cells and perform a drug permeability test using this model.

pH-bioresponsive poly(ε-caprolactone)-based polymersome for effective drug delivery in cancer and protein glycoxidation prevention.

Artificial nanostructures using polymers to produce polymeric vesicles are inspired by the many intricate structures found in living organisms. Polymersomes are a class of self-assembled vesicles known for their great stability and application in drug delivery. They can be tuned according to their intended use by changing their components and introducing activable block copolymers that transform these polymersomes into smart nanocarriers. In this study, we propose the synthesis of a poly (ethylene oxide)-poly (ε-caprolactone)-based polymersome (PEO-PCL) loaded with GSH as a pH-responsive drug delivery molecule for cancer and protein alteration inhibition. Initially, the nanocarrier was synthesized and characterized by DLS, TEM/SEM microscopy as well as gel permeation chromatography (GPC) and 1H NMR. Their CMC formation, encapsulation efficiency, and pH responsiveness were analyzed. In addition, empty and GSH-loaded PEO-PCL polymersomes were tested for their toxicity and therapeutic effect on normal and cancer cells via an MTT test. Subsequently, protein alteration models (aggregation, glycation, and oxidation) were performed in vitro where the polymersomes were tested. Results showed that other than being non-toxic and able to highly encapsulate and release the GSH in response to acidic conditions, the nanocomposites do not hinder its content's ameliorative effects on cancer cells and protein alterations. This infers that polymeric nanocarriers can be a base for future smart biomedicine applications and theranostics.

Coculture model of blood-brain barrier on electrospun nanofibers.

The blood-brain barrier (BBB) is a control mechanism that limits the diffusion of many substances to the central nervous system (CNS). In this study, we designed an in-vitro 3-dimensional BBB system to obtain a fast and reliable model to mimic drug delivery characteristics of the CNS. A support membrane of polycaprolactone nanofiber surfaces was prepared using electrospinning. After confirming the fiber morphology and size, endothelial cells (HUVEC) and glial cells were cultured on either side of this membrane. The model's similarity to in vivo physiology was tested with a home-designed transmembrane resistance (TR) device, with positive and negative control molecules. Finally, 2 doses of methotrexate (MTX), a chemotherapy agent, were applied to the model, and its permeability through the model was determined indirectly by a vitality test on the MCF-7 cell line. Nicotine, the positive control, completed its penetration through the model almost instantly, while albumin, the negative control, was blocked significantly even after 2 days. MTX reached a deadly threshold 24 h after application. The TR value of the model was promising, being around 260 ohm.cm2. The provided model proposes a disposable and reliable tool for investigating drug permeability through the BBB and has the potential to reduce the number of animal experiments.

pH-Responsive Polymersome Microparticles as Smart Cyclodextrin-Releasing Agents.

Cyclodextrins (CDs) are increasingly drawing attention as potential therapeutic tools in the treatment of cholesterol-associated diseases. However, bioavailability and delivery of CDs in the monomeric form still remain challenging. CD-based macromolecular systems seem to display a promising capacity in overcoming some of these limitations. Therefore, smart, stimuli-responsive nanosystems are currently being investigated in order to provide improved CD-releasing agents. Herein, we present a novel class of CD-based polymersome microparticles (CD-PMs) designed for potential therapeutic use. A new synthetic route to obtain a CD-appended, pH-sensitive polymer that self-assembles into a stable polymersome microparticle is reported. Through an easy-to-use approach, a benzoic imine bond is incorporated into a poly(ε-caprolactone) backbone and employed as a building block in the construction of the nanoarchitecture. The CD-PMs show cellular uptake representing a promising potential therapeutic tool in the treatment of cholesterol-associated conditions such as neurodegenerative diseases.

Mechanobiology of cells and cell systems, such as organoids.

Organoids are in vitro 3D self-organizing tissues that mimic embryogenesis. Organoid research is advancing at a tremendous pace, since it offers great opportunities for disease modeling, drug development and screening, personalized medicine, as well as understanding organogenesis. Mechanobiology of organoids is an unexplored area, which can shed light to several unexplained aspects of self-organization behavior in organogenesis. It is becoming evident that collective cell behavior is distinctly different from individual cells' conduct against certain stimulants. Inherently consisting of higher number of degrees of freedom for cell motility and more complex cell-to-cell and cell-to-extracellular matrix behavior, understanding mechanotransduction in organoids is even more challenging compared with cell communities in 2D culture conditions. Yet, deciphering mechanobiology of organoids can help us understand effects of mechanical cues in health and disease, and translate findings of basic research toward clinical diagnosis and therapy.

The use of bacterial cellulose as a basement membrane improves the plausibility of the static in vitro blood-brain barrier model.

There are several blood-brain barrier (BBB) models available for pharmaceutical research, but none of those are able to properly imitate the permeability of this special barrier. In this study, it is aimed to produce different BBB models with different cellular combinations and different basement membrane polymers, such as polyethylene terephthalate (PET) and bacterial cellulose (BC), which has not been used for BBB models before, to compare their barrier properties. Primary human brain microvascular endothelial cells were seeded on the luminal side and primary human astrocytes and/or primary human brain microvascular pericytes were seeded on the abluminal side of the membranes. Immunofluorescence (IF) staining results indicate that the expression of tight and adherence junction proteins increases on the 5th day of the cultivation. In accordance with Live-Dead staining results, IF images show that cells in the model lose their viability before the 10th day. Transendothelial electrical resistance (TEER) measurements indicate that BC membrane leads to statistically higher (p < 0.05) TEER values than the standard Transwell PET insert membrane. Sucrose and caffeine permeability values of all models are close to in vivo values. BC shows potential to be used as a more reliable basement membrane for BBB models for pharmaceutical research.

Optimization of bacterial cellulose production by Gluconacetobacter xylinus using carob and haricot bean.

Bacterial cellulose (BC) can be used in medical, biomedical, electronic, food, and paper industries because of its unique properties distinguishing it from plant cellulose. BC production was statistically optimized by Gluconacetobacter xylinus strain using carob and haricot bean (CHb) medium. Eight parameters were evaluated by Plackett-Burman Design and significant three parameters were optimized by Central Composite Design. Optimal conditions for production of BC in static culture were found as: 2.5g/L carbon source, 2.75g/L protein source, 9.3% inoculum ratio, 1.15g/L citric acid, 2.7g/L Na2HPO4, 30°C incubation temperature, 5.5 initial pH, and 9days of incubation. This study reveals that BC production can be carried out using carob and haricot bean extracts as carbon and nitrogen sources, and CHb medium has higher buffering capacity compared to Hestrin and Schramm media. Model obtained from this study is used to predict and optimize BC production yield using CHb medium.

The effects of different intensities, frequencies and exposure times of extremely low-frequency electromagnetic fields on the growth of Staphylococcus aureus and Escherichia coli O157:H7.

The impact of different types of extremely low-frequency electromagnetic fields (ELF-EMF) on the growth of Staphylococcus aureus and Escherichia coli O157:H7 was investigated. The cultures of bacteria in broth media were exposed to sinusoidal homogenous ELF-EMF with 2 and 4 mT magnetic intensities. Each intensity for each bacteria was combined with three different frequencies (20, 40 and 50 Hz), and four different exposure times (1, 2, 4 and 6 h). A cell suspension of each experiment was diluted for the appropriate range and inoculated to Mueller-Hinton Agar (MHA) plates after exposure to ELF-EMF. The number of colony forming units (CFU) of both strains was obtained after incubation at 37 °C for 24 h. Data were statistically evaluated by one-way analysis of variance (ANOVA), statistical significance was described at p < 0.05 and data were compared with their non-exposed controls. Magnetic intensity, frequency and exposure time of ELF-EMFs changed the characteristic responses for both microorganisms. Samples exposed to ELF-EMF showed a statistically significant decrease compared to their controls in colony forming capability, especially at long exposure times. An exposure to 4 mT-20 Hz ELF-EMF of 6 h produced maximum inhibition of CFU compared to their controls for both microorganisms (95.2% for S. aureus and 85% for E. coli).