Magnetic-Responsive Doxorubicin-Containing Materials Based on Fe3O4 Nanoparticles with a SiO2/PEG Shell and Study of Their Effects on Cancer Cell Lines

Novel nanocomposite materials based on Fe3O4 magnetic nanoparticles (MNPs) coated with silica and covalently modified by [(3-triethoxysilyl)propyl]succinic acid–polyethylene glycol (PEG 3000) conjugate, which provides a high level of doxorubicin (Dox) loading, were obtained. The efficiency of Dox desorption from the surface of nanomaterials under the action of an alternating magnetic field (AMF) in acidic and neutral media was evaluated. Their high cytotoxicity against tumor cells, as well as the drug release upon application of AMF, which leads to an increase in the cytotoxic effect, was demonstrated

Optimizing the Composition of the Substrate Enhances the Performance of Peroxidase-like Nanozymes in Colorimetric Assays: A Case Study of Prussian Blue and 3,3′-Diaminobenzidine

One of the emerging trends in modern analytical and bioanalytical chemistry involves the substitution of enzyme labels (such as horseradish peroxidase) with nanozymes (nanoparticles possessing enzyme-like catalytic activity). Since enzymes and nanozymes typically operate through different catalytic mechanisms, it is expected that optimal reaction conditions will also differ. The optimization of substrates for nanozymes usually focuses on determining the ideal pH and temperature. However, in some cases, even this step is overlooked, and commercial substrate formulations designed for enzymes are utilized. This paper demonstrates that not only the pH but also the composition of the substrate buffer, including the buffer species and additives, significantly impact the analytical signal generated by nanozymes. The presence of enhancers such as imidazole in commercial substrates diminishes the catalytic activity of nanozymes, which is demonstrated herein through the use of 3,3′-diaminobenzidine (DAB) and Prussian Blue as a model chromogenic substrate and nanozyme. Conversely, a simple modification to the substrate buffer greatly enhances the performance of nanozymes. Specifically, in this paper, it is demonstrated that buffers such as citrate, MES, HEPES, and TRIS, containing 1.5–2 M NaCl or NH4Cl, substantially increase DAB oxidation by Prussian Blue and yield a higher signal compared to commercial DAB formulations. The central message of this paper is that the optimization of substrate composition should be an integral step in the development of nanozyme-based assays. Herein, a step-by-step optimization of the DAB substrate composition for Prussian Blue nanozymes is presented. The optimized substrate outperforms commercial formulations in terms of efficiency. The effectiveness of the optimized DAB substrate is affirmed through its application in several commonly used immunostaining techniques, including tissue staining, Western blotting assays of immunoglobulins, and dot blot assays of antibodies against SARS-CoV-2

Synthesis of Prussian Blue nanoparticles in water/alcohol mixtures

Prussian Blue, a blue coordination polymer, emerges as a promising candidate in the realm of biomedicine. Its nanoparticles, known as catalytic labels or nanozymes, exhibit remarkable peroxidase-like properties and serve as effective antioxidants. Unsurprisingly, the demand for synthesizing Prussian Blue nanoparticles with customizable sizes is on the rise. In this study, we unveil a novel approach to synthesizing Prussian Blue nanoparticles. In this work, the synthesis of Prussian Blue nanoparticles by reducing an equimolar mixture of FeCl3 and K3[Fe(CN)6] with hydrogen peroxide in different water-alcohol mixtures was demonstrated for the first time. Alcohols with a lower dielectric constant (propanol-1, isopropyl alcohol, and tert-butanol) contribute to an increase in nanoparticle size, particularly at mole fractions of 0.02-0.05 and beyond. Conversely, alcohols with a higher dielectric constant (ethanol, methanol, ethylene glycol, and propylene glycol, excluding glycerol) demonstrate the ability to decrease nanoparticle size at mole fractions of 0.2-0.26 and higher. Building upon these findings, we present a scalable and reproducible method for preparing small Prussian Blue nanoparticles, measuring 30-40 nm, with enhanced peroxidase-like activity using 79.2% ethylene glycol as a solvent. The proposed mechanism behind the effect of ethylene glycol involves the limitation of both growth and secondary aggregation of Prussian Blue nanoparticles. These synthesized nanoparticles prove their efficiency as catalytic labels in a model vertical flow immunoassay designed to detect antibodies against SARS-CoV-2

Prussian blue nanozymes with enhanced catalytic activity: size tuning and application in ELISA-like immunoassay

Prussian blue nanozymes possessing peroxidase-like activity gather significant attention as alternatives to natural enzymes in therapy, biosensing, and environmental remediation. Recently, prussian blue nanoparticles with enhanced catalytic activity prepared by reduction of FeCl3/K3[Fe(CN)6] mixture have been reported. These nanoparticles were denoted as ‘artificial peroxidase’ nanozymes. Our study provides insights into the process of synthesis of ‘artificial peroxidase’ nanozymes. We studied how the size of nanozymes and synthesis yield can be controlled via adjustment of the synthesis conditions. Based on these results, we developed a reproducible and scalable method for the preparation of ‘artificial peroxidase’ with tunable sizes allowing the obtaining of nanozymes with enhanced catalytic activity. ‘Artificial peroxidase’ nanozymes modified with gelatin shell and functionalized with affine molecules were applied as labels in colorimetric immunoassays of prostate-specific antigen and tetanus antibodies, enabling detection of these analytes in the range of clinically relevant concentrations. Protein coating provides excellent colloidal stability of nanozymes in physiological conditions and stability upon long-term storage

Dynamic Transition from α‑Helices to β‑Sheets in Polypeptide Coiled-Coil Motifs

We carried out dynamic force manipulations in silico on a variety of coiled-coil protein fragments from myosin, chemotaxis receptor, vimentin, fibrin, and phenylalanine zippers that vary in size and topology of their α-helical packing. When stretched along the superhelical axis, all superhelices show elastic, plastic, and inelastic elongation regimes and undergo a dynamic transition from the α-helices to the β-sheets, which marks the onset of plastic deformation. Using the Abeyaratne-Knowles formulation of phase transitions, we developed a new theoretical methodology to model mechanical and kinetic properties of protein coiled-coils under mechanical nonequilibrium conditions and to map out their energy landscapes. The theory was successfully validated by comparing the simulated and theoretical force-strain spectra. We derived the scaling laws for the elastic force and the force for α-to-β transition, which can be used to understand natural proteins’ properties as well as to rationally design novel biomaterials of required mechanical strength with desired balance between stiffness and plasticity

Magnetism and EPR Spectroscopy of Nanocrystalline and Amorphous TiO₂: Fe upon Al Doping

This work is devoted to the study of the magnetic properties and Electron Paramagnetic Resonance (EPR) spectroscopy of TiO2:Fe nanoparticles doped with Al in different structural states. The sol-gel methods have been used to obtain the particles in both crystalline (average size from 3 to 20 nm) and X-ray amorphous states. The electron paramagnetic resonance spectra of crystalline samples TiO2:Fe doped with aluminum besides a resonance line with g-factor ~2 exhibit a small signal with a g-factor of 4.3 from Fe3+ ions with rhombohedral distortions. The fraction of Fe3+ with rhombohedral distortions increases with increasing aluminum content. For the amorphous state at Al doping, the resonance with a g-factor of 4.3 is completely dominant in the electron paramagnetic resonance spectrum. The density functional theory calculation shows that aluminum prefers to be localized near iron ions, distorting the nearest Fe3+ environment. The complex integral electron paramagnetic resonance spectrum of all samples was fitted with sufficient accuracy by three separate resonance lines with different widths and intensities. The temperature behavior of the electron paramagnetic resonance spectrum can be described by the coexistence of paramagnetic centers (isolated Fe3+ ions including dipole-dipole interactions) and iron clusters with negative exchange interactions

Computer vision vs. spectrofluorometer-assisted detection of common nitro-explosive components with bola-type PAH-based chemosensors

Computer vision (CV) algorithms are widely utilized in imaging processing for medical and personal electronics applications. In sensorics CV can provide a great potential to quantitate chemosensors' signals. Here we wish to describe a method for the CV-assisted spectrofluorometer-free detection of common nitro-explosive components, e.g. 2,4-dinitrotoluene (DNT) and 2,4,6-trinitrotoluene (TNT), by using polyaromatic hydrocarbon (PAH, PAH = 1-pyrenyl or 9-anthracenyl) – based bola-type chemosensors. The PAH components of these chemical bolas are able to form stable, bright emissive in a visual wavelength region excimers, which allows their use as extended matrices of the RGB colors after imaging and digital processing. In non-polar solvents, the excimers have poor chemosensing properties, while in aqueous solutions, due to the possible micellar formation, these excimers provide “turn-off” fluorescence detection of DNT and TNT in the sub-nanomolar concentrations. A combination of these PAH-based fluorescent chemosensors with the proposed CV-assisted algorithm offers a fast and convenient approach for on-site, real-time, multi-thread analyte detection without the use of fluorometers. Although we focus on the analysis of nitro-explosives, the presented method is a conceptual work describing a general use of CV for quantitative fluorescence detection of various analytes as a simpler alternative to spectrofluorometer-assisted methods

Magnetism and Electronic State of Iron Ions on the Surface and in the Core of TiO₂ Nanoparticles

In this paper, the electron and magnetic state of iron placed either on the surface or in the core of TiO2 nanoparticles were investigated using magnetometric methods, electron paramagnetic resonance (EPR) and Mössbauer spectroscopy. It was demonstrated that the EPR spectra of TiO2 samples with iron atoms localized both on the surface and in the core of specific features depending on the composition and size of the nanoparticles. Theoretical calculations using the density functional theory (DFT) method demonstrated that the localization of Fe atoms on the surface is characterized by a considerably larger set of atomic configurations as compared to that in the core of TiO2 nanoparticles. Mössbauer spectra of the samples doped with Fe atoms both on the surface and in the core can be described quite satisfactorily using two and three doublets with different quadrupole splitting, respectively. This probably demonstrates that the Fe atoms on particle surface and in the bulk are in different unlike local surroundings. All iron ions, both on the surface and in the core, were found to be in the Fe3+ high-spin state

Versatile Reactivity of Mn-II Complexes in Reactions with N-Donor Heterocycles: Metamorphosis of Labile Homometallic Pivalates vs. Assembling of Endurable Heterometallic Acetates

International audienceReaction of 2,2'-bipyridine (2,2'-bipy) or 1,10-phenantroline (phen) with [Mn(Piv)(EtOH)] led to the formation of binuclear complexes [Mn(Piv)L] (L = 2,2'-bipy (), phen (); Piv is the anion of pivalic acid). Oxidation of or by air oxygen resulted in the formation of tetranuclear Mn complexes [MnO(Piv)L] (L = 2,2'-bipy (), phen ()). The hexanuclear complex [Mn(OH)(Piv)(pym)] () was formed in the reaction of [Mn(Piv)(EtOH)] with pyrimidine (pym), while oxidation of produced the coordination polymer [MnO(Piv)(pym)] (). Use of pyrazine (pz) instead of pyrimidine led to the 2D-coordination polymer [Mn(OH)(Piv)(µ-pz)] (). Interaction of [Mn(Piv)(EtOH)] with FeCl resulted in the formation of the hexanuclear complex [MnFeO(Piv)(MeCN)(HPiv)] (). The reactions of [MnFeO(OAc)(HO)] with 4,4'-bipyridine (4,4'-bipy) or -1,2-(4-pyridyl)ethylene (bpe) led to the formation of 1D-polymers [MnFeO(OAc)L]·2DMF, where L = 4,4'-bipy (·2DMF), bpe (·2DMF) and [MnFeO(OAc)(bpe)(DMF)]·3.5DMF (·3.5DMF). All complexes were characterized by single-crystal X-ray diffraction. Desolvation of ·3.5DMF led to a collapse of the porous crystal lattice that was confirmed by PXRD and N sorption measurements, while alcohol adsorption led to porous structure restoration. Weak antiferromagnetic exchange was found in the case of binuclear Mn complexes ( = -1.03 cm for and ). According to magnetic data analysis ( = -(2.69 ÷ 0.42) cm) and DFT calculations ( = -(6.9 ÷ 0.9) cm) weak antiferromagnetic coupling between Mn ions also occurred in the tetranuclear {Mn(OH)(Piv)} unit of the 2D polymer . In contrast, strong antiferromagnetic coupling was found in oxo-bridged trinuclear fragment {MnFeO(OAc)} in ·3.5DMF ( = -57.8 cm, = -20.12 cm)