Pavel Veverka

Mechanism of hypercrosslinking of chloromethylated styrene–divinylbenzene copolymers

Abstract Hypercrosslinked polymers are remarkable materials exhibiting extremely high apparent surface area and exceptional sorption properties. Their special morphology is a result of the fixation of expanded position of polymer chains in a good (swelling) solvent by methylene crosslinking bridges created by the Friedel–Crafts reaction of chloromethyl groups. We have performed a detailed investigation of the formation of the crosslinking bridges in the reaction starting from the swollen, already chloromethylated polymer. Conversion of the chloromethyl groups was determined from the residual chlorine content. The total chlorine content determined after burning a sample of the polymer in oxygen and the content of reactive, accessible chlorine determined by reaction of the chloromethyl groups with pyridine were differentiated. Changes in the content of both types of chlorine were monitored for varying reaction conditions (time, catalyst content, etc.) and were correlated with the development of the microporous texture resulting from the hypercrosslinking of the swollen polymer matrix.

Magnetic heating by cobalt ferrite nanoparticles

In the quest for suitable materials for hyperthermia we explored the preparation and properties of nanoparticles of Co ferrite. The material was produced by coprecipitation from water solution of Co and Fe chlorides and afterwards annealed at 400, 600 and 800 °C. The resulting particles were characterized by XRD, TEM, Mossbauer spectroscopy, and dc and ac magnetometry. The heating experiments in ac magnetic fields of various amplitudes were performed with diluted systems of particles suspended in agarose gel and the results were interpreted on the basis of the ac magnetic losses measured at various temperatures. The increase of magnetic losses and consequently of the heating efficiency with increasing temperature is explained by the strong dependence of the constant of magnetocrystalline anisotropy of Co ferrite on temperature.

Silica encapsulated manganese perovskite nanoparticles for magnetically induced hyperthermia without the risk of overheating

Nanoparticles of manganese perovskite of the composition La(0.75)Sr(0.25)MnO(3) uniformly coated with silica were prepared by encapsulation of the magnetic cores (mean crystallite size 24 nm) using tetraethoxysilane followed by fractionation. The resulting hybrid particles form a stable suspension in an aqueous environment at physiological pH and possess a narrow hydrodynamic size distribution. Both calorimetric heating experiments and direct measurements of hysteresis loops in the alternating field revealed high specific power losses, further enhanced by the encapsulation procedure in the case of the coated particles. The corresponding results are discussed on the basis of complex characterization of the particles and especially detailed magnetic measurements. Moreover, the Curie temperature (335 K) of the selected magnetic cores resolves the risk of local overheating during hyperthermia treatment.

Dual imaging probes for magnetic resonance imaging and fluorescence microscopy based on perovskite manganite nanoparticles

The present study reveals the potential of magnetic nanoparticles based on the La0.75Sr0.25MnO3 perovskite manganite for magnetic resonance imaging (MRI). Moreover, it describes the development of the dual imaging probe where the magnetic cores are combined with a fluorescent moiety while the improved colloidal stability is achieved by a two-ply silica shell. At first, the magnetic cores of La0.75Sr0.25MnO3 are coated with a hybrid silica layer, comprising a covalently attached fluorescein moiety that is subsequently covered by a pure silica layer providing the enhanced stability. The detailed characterization of the intermediate and the final product reveals the importance of the complex two-ply shell. Viability tests show that the complete particles are suitable for biological studies. Internalization of the particles and their presence in intracellular vesicles are observed by fluorescence microscopy in different cell types. Further experiments prove no fatal interference with the vitality and insulin releasing ability of labeled pancreatic islets. Relaxometric measurements confirm high spin–spin relaxivities at magnetic fields of B0 = 0.5–3 T, while visualisation of in vitro labeled pancreatic islets by MRI is successfully tested.

Core–shell La1−xSrxMnO3 nanoparticles as colloidal mediators for magnetic fluid hyperthermia

Core–shell nanoparticles consisting of La0.75Sr0.25MnO3 cores covered by silica were synthesized by a procedure consisting of several steps, including the sol–gel method in the presence of citric acid and ethylene glycol, thermal and mechanical treatment, encapsulation employing tetraethoxysilane and final separation by centrifugation in order to get the required size fraction. Morphological studies revealed well-separated particles that form a stable water suspension. Magnetic studies include magnetization measurements and investigation of the ferromagnetic–superparamagnetic–paramagnetic transition. Magnetic heating experiments in ‘calorimetric mode’ were used to determine the heating efficiency of the particles in water suspension and further employed for biological studies of extracellular and intracellular effects analysed by tests of viability.

Silica‐coated La0.75Sr0.25MnO3 nanoparticles for magnetically driven DNA isolation

Magnetic La(0.75)Sr(0.25)MnO(3) nanoparticles possessing an approximately 20-nm-thick silica shell (LSMO(0.25)@SiO(2) ) were characterised and tested for the isolation of PCR-ready bacterial DNA. The results presented here show that the nanoparticles do not interfere in PCR. DNA was apparently reversibly adsorbed on their silica shell from the aqueous phase system (16% PEG 6000-2 M NaCl). The method proposed was used for DNA isolation from complex food samples (dairy products and probiotic food supplements). The isolated DNA was compatible with PCR. The main advantages of the nanoparticles tested for routine use were their high colloidal stability allowing a more precise dosage and therefore high reproducibility of DNA isolation.

Silica-coated manganite and Mn-based ferrite nanoparticles: a comparative study focused on cytotoxicity

Magnetic oxide nanoparticles provide a fascinating tool for biological research and medicine, serving as contrast agents, magnetic carriers, and core materials of theranostic systems. Although the applications rely mostly on iron oxides, more complex oxides such as perovskite manganites may provide a much better magnetic performance. To assess the risk of their potential use, in vitro toxicity of manganite nanoparticles was thoroughly analysed and compared with another prospective system of Mn–Zn ferrite nanoparticles. Magnetic nanoparticles of La0.63Sr0.37MnO3 manganite were prepared by two distinct methods, namely the molten salt synthesis and the traditional sol–gel route, whereas nanoparticles of Mn0.61Zn0.42Fe1.97O4 ferrite, selected as a comparative material, were synthesized by a new procedure under hydrothermal conditions. Magnetic cores were coated with silica and, moreover, several samples of manganite nanoparticles with different thicknesses of silica shell were prepared. The size-fractionated and purified products were analysed using transmission electron microscopy, dynamic light scattering, measurement of the zeta-potential dependence on pH, IR spectroscopy, and SQUID magnetometry. The silica-coated products with accurately determined concentration by atomic absorption spectroscopy were subjected to a robust evaluation of their cytotoxicity by four different methods, including detailed analysis of the concentration dependence of toxicity, analysis of apoptosis, and experiments on three different cell lines. The results, comparing two manganese-containing systems, clearly indicated superior properties of the Mn–Zn ferrite, whose silica-coated nanoparticles show very limited toxic effects and thus constitute a promising material for bioapplications.

Clusters of Magnetic Nanoparticles as Contrast Agents for MRI: Effect of Aggregation on Transverse Relaxivity

The effect of aggregation of magnetic nanoparticles on the transverse relaxivity (r2) is analyzed with respect to the size of clusters. The nanoparticles employed are based on La0.75Sr0.25MnO3 ferromagnetic phase with the mean size of crystallites dXRD = 26 nm synthetized via sol-gel route followed by thermal treatment and mechanical processing. The subsequent silica coating provides colloidally stable particles whose magnetic cores are mostly composed of compact clusters of manganite crystallites. The product has been subjected to repeated differential centrifugation and several size fractions, possessing the same dXRD but differing in the effective size of magnetic cores, are isolated. Thorough analyses of their size distributions by transmission electron microscopy and dynamic light scattering measurements are carried out together with SQUID magnetometry. The concentration of particles in aqueous suspensions is accurately determined by atomic absorption spectroscopy and a detailed study of transverse relaxation at the magnetic field B0 = 0.5 T is performed. The highest r2 values are clearly observed for midsized clusters and the temperature dependence of r2 resembles the evolution of magnetization with temperature. Supplemental samples with different thicknesses of the silica shell are also synthetized and thoroughly analyzed.

The impact of silica encapsulated cobalt zinc ferrite nanoparticles on DNA, lipids and proteins of rat bone marrow mesenchymal stem cells.

Abstract Nanomaterials are currently the subject of intense research due to their wide variety of potential applications in the biomedical, optical and electronic fields. We prepared and tested cobalt zinc ferrite nanoparticles (Co0.5Zn0.5Fe2O4+γ [CZF-NPs]) encapsulated by amorphous silica in order to find a safe contrast agent and magnetic label for tracking transplanted cells within an organism using magnetic resonance imaging (MRI). Rat mesenchymal stem cells (rMSCs) were labeled for 48 h with a low, medium or high dose of CZF-NPs (0.05; 0.11 or 0.55 mM); silica NPs (Si-NPs; 0.11 mM) served as a positive control. The internalization of NPs into cells was verified by transmission electron microscopy. Biological effects were analyzed at the end of exposure and after an additional 72 h of cell growth without NPs. Compared to untreated cells, Annexin V/Propidium Iodide labeling revealed no significant cytotoxicity for any group of treated cells and only a high dose of CZF-NPs slowed down cell proliferation and induced DNA damage, manifested as a significant increase of DNA-strand breaks and oxidized DNA bases. This was accompanied by high concentrations of 15-F2t-isoprostane and carbonyl groups, demonstrating oxidative injury to lipids and proteins, respectively. No harmful effects were detected in cells exposed to the low dose of CZF-NPs. Nevertheless, the labeled cells still exhibited an adequate relaxation rate for MRI in repeated experiments and ICP-MS confirmed sufficient magnetic label concentrations inside the cells. The results suggest that the silica-coated CZF-NPs, when applied at a non-toxic dose, represent a promising contrast agent for cell labeling.

The effect of magnetic nanoparticles on neuronal differentiation of induced pluripotent stem cell-derived neural precursors

Introduction Magnetic resonance (MR) imaging is suitable for noninvasive long-term tracking. We labeled human induced pluripotent stem cell-derived neural precursors (iPSC-NPs) with two types of iron-based nanoparticles, silica-coated cobalt zinc ferrite nanoparticles (CZF) and poly-l-lysine-coated iron oxide superparamagnetic nanoparticles (PLL-coated γ-Fe2O3) and studied their effect on proliferation and neuronal differentiation. Materials and methods We investigated the effect of these two contrast agents on neural precursor cell proliferation and differentiation capability. We further defined the intracellular localization and labeling efficiency and analyzed labeled cells by MR. Results Cell proliferation was not affected by PLL-coated γ-Fe2O3 but was slowed down in cells labeled with CZF. Labeling efficiency, iron content and relaxation rates measured by MR were lower in cells labeled with CZF when compared to PLL-coated γ-Fe2O3. Cytoplasmic localization of both types of nanoparticles was confirmed by transmission electron microscopy. Flow cytometry and immunocytochemical analysis of specific markers expressed during neuronal differentiation did not show any significant differences between unlabeled cells or cells labeled with both magnetic nanoparticles. Conclusion Our results show that cells labeled with PLL-coated γ-Fe2O3 are suitable for MR detection, did not affect the differentiation potential of iPSC-NPs and are suitable for in vivo cell therapies in experimental models of central nervous system disorders.