Věra Kašpárková

Stem cell differentiation on conducting polyaniline

Polyaniline is a promising conducting polymer with broad application potential in biomedicine. Its medical use, however, requires both biocompatibility and suitable physico-chemical and surface properties. The microstructure, electrical properties, and surface characteristics of polyaniline salt, polyaniline base, and polyaniline deposited with biologically active poly(2-acrylamido-2-methyl-1-propanesulfonic acid) were revealed using atomic force microscopy, contact angle measurements, and Raman spectroscopy. As conducting polymers can be preferentially applied in tissue engineering of heart and nervous tissues, the cardiomyogenesis in pure cardiomyocytes derived from embryonic stem cells and neurogenesis in neural progenitors isolated from embryonal 13 dpc brain were further investigated. The results show that neither cardiomyogenesis nor neurogenesis were influenced by any of the tested polyaniline films. However, the most favorable cell behaviour was observed on pristine polyaniline base; therefore, polyaniline in pristine forms without any further modification can be applied in a variety of biomedical fields.

Blood coagulation and platelet adhesion on polyaniline films

Polyaniline is a promising conducting polymer with still increasing application potential in biomedicine. Its surface modification can be an efficient way how to introduce desired functional groups and to control its properties while keeping the bulk characteristics of the material unchanged. The purpose of the study was to synthetize thin films of pristine conducting polyaniline hydrochloride, non-conducting polyaniline base and polyaniline modified with poly(2-acrylamido-2-methyl-1-propanesulfonic acid) (PAMPSA) and investigate chosen parameters of their hemocompatibility. The modification was performed either by introduction of PAMPSA during the synthesis or by reprotonation of polyaniline base. The polyaniline hydrochloride and polyaniline base had no impact on blood coagulation and platelet adhesion. By contrast, the polyaniline reprotonated with PAMPSA completely hindered coagulation thanks to its interaction with coagulation factors Xa, Va and IIa. The significantly lower platelets adhesion was also found on this surface. Moreover, this film maintains its conductivity at pH of 6, which is an improvement in comparison with standard polyaniline hydrochloride losing most of its conductivity at pH of 4. Polyaniline film with PAMPSA introduced during synthesis had an impact on platelet adhesion but not on coagulation. The combined conductivity, anticoagulation activity, low platelet adhesion and improved conductivity at pH closer to physiological, open up new possibilities for application of polyaniline reprotonated by PAMPSA in blood-contacting devices, such as catheters or blood vessel grafts.

Characteristics of silver nanoparticles in vehicles for biological applications

Silver nanoparticles (AgNPs) have been used for decades as anti-bacterial agents in various industrial fields such as cosmetics, health industry, food storage, textile coatings and environmental applications, although their toxicity is not fully recognized yet. Antimicrobial and catalytic activity of AgNPs depends on their size as well as structure, shape, size distribution, and physico-chemical environment. The unique properties of AgNPs require novel or modified toxicological methods for evaluation of their toxic potential combined with robust analytical methods for characterization of nanoparticles applied in relevant vehicles, e.g., culture medium with/without serum and phosphate buffered saline.

Behaviour of silver nanoparticles in simulated saliva and gastrointestinal fluids

Continuously increasing application of silver nanoparticles (AgNPs) requires information on their safety and performance under biological conditions. Assessment of AgNPs in biological systems is also related to availability of robust toxicological methods for evaluation of toxic potential of AgNPs and information on their physicochemical state. Silver nanoparticles were subjected to action of simulated saliva, gastric and intestinal fluids, appropriately supplemented with digestive enzymes pepsin or pancreatin. The behaviour of AgNPs was determined using dynamic light scattering and transmission electron microscopy, and their toxicity as well as capability to induce inflammatory reactions were assessed using reconstructed human tissue models (EpiOral, EpiGingival, EpiIntestinal). The study revealed that during exposure to the fluids, AgNPs size and morphology changed and depended on composition and pH of the respective fluid. If present, the change in terms of growth of AgNPs size occurred immediately after contact of AgNPs with the respective fluid and continued with prolonged time of contact. A pilot study on reconstituted human tissue models revealed low toxicity and inflammatory effects of AgNPs and confirmed the suitability of 3-D models for toxicological studies including bioavailability.

DEVELOPMENT OF AN HPLC METHOD FOR THE DETERMINATION OF GLYCEROL OXIDATION PRODUCTS

An HPLC method for the separation of glycerol oxidation products, namely glyceraldehyde, dihydroxyacetone, mesoxalic, tartronic, and glycolic and glyceric acids on an ion-exchange 8% cross-linked calcium sulfonated divinylbenzene-styrene resin column was developed and validated. The conditions reported include temperature (70°C), flow rate (0.5 mL/min) and concentration of the mobile phase (3 mM H2SO4) using isocratic elution with ultraviolet and refractometric detectors. The effect of the mobile phase flow rate and concentration as well as column temperature on the resolution of peaks is described. Excellent correlation coefficient in the calibration model was observed for all analytes over the concentration range of 0.5 to 10 mg/mL. The method was also validated in terms of intra-day precision, sensitivity, accuracy, and detection and quantification limits. The method conditions were applied to the identification of products derived from the chemical oxidation of glycerol. Supplemental materials are available for this article. Go to the publishers online edition of the Journal of Liquid Chromatography & Related Technologies to view the supplemental file.

On the Precision of Particle Size Analysis by Micro‐Thermal Field‐Flow Fractionation

Abstract Micro‐Thermal Field‐Flow Fractionation of polymer colloidal particles was performed in two different laboratories. Short term repeatability of the experimental retentions obtained in a single laboratory has been found to be very high. The short term precision (expressed as percent standard deviation) of the determination of average particle diameter can reach the values greater than 1%, relative. Average repeatability of the retentions in both laboratories was better than 3%, relative when using identical experimental protocol. No other method of particle size analysis can provide the results of a comparable precision. Average repeatability of the width of the raw fractograms, which contains the information on particle size distribution, is of the order of 5%, relative. However, this value cannot be considered as the ultimate limit because the experiments were not carried at the low flow rate of the carrier liquid permitted to reach much higher resolution. The effect of the stability of the most important operational variables, such as the temperature drop between the cold and hot walls, the temperature of the cold wall, and the flow rate of the carrier liquid, on the precision of the analytical results is discussed. This article is dedicated to the memory of J. C. Giddings (1930–1996)

Formulation, Characterization and Properties of Hemp Seed Oil and Its Emulsions

The formulation, characterization, and anticipated antibacterial properties of hemp seed oil and its emulsions were investigated. The oil obtained from the seeds of Cannabis sativa L. in refined and unrefined form was characterized using iodine, saponification, acid values, and gas chromatography, and was employed for the preparation of stable oil-in-water emulsions. The emulsions were prepared using pairs of non-ionic surfactants (Tween, Span). The effects of the emulsification method (spontaneous emulsification vs. high-intensity stirring), hydrophilic lipophilic balance (HLB), type and concentration of surfactant, and oil type on the size and distribution of the emulsion particles were investigated. It was found that the ability to form stable emulsions with small, initial particle sizes is primarily dependent on the given method of preparation and the HLB value. The most efficient method of emulsification that afforded the best emulsions with the smallest particles (151 ± 1 nm) comprised the high-energy method, and emulsions stable over the long-term were observed at HBL 9 with 10 wt % concentration of surfactants. Under high-intensity emulsification, refined and unrefined oils performed similarly. The oils as well as their emulsions were tested against the growth of selected bacteria using the disk diffusion and broth microdilution methods. The antibacterial effect of hemp seed oil was documented against Micrococcus luteus and Staphylococcus aureus subsp. aureus. The formulated emulsions did not exhibit the antibacterial activity that had been anticipated.

Cell-compatible conducting polyaniline films prepared in colloidal dispersion mode

Conducting polyaniline can be prepared and modified using several procedures, all of which can significantly influence its applicability in different fields of biomedicine or biotechnology. The modifications of surface properties are crucial with respect to the possible applications of this polymer in tissue engineering or as biosensors. Innovative technique for preparing polyaniline films via in-situ polymerization in colloidal dispersion mode using four stabilizers (poly-N-vinylpyrrolidone; sodium dodecylsulfate; Tween 20 and Pluronic F108) was developed. The surface energy, conductivity, spectroscopic features, and cell compatibility of thin polyaniline films were determined using contact-angle measurement, the van der Pauw method, Fourier-transform infrared spectroscopy, and assay conducted on mouse fibroblasts, respectively. The stabilizers significantly influenced not only the surface and electrical properties of the films but also their cell compatibility. Sodium dodecylsulfate seems preferentially to combine both the high conductivity and good cell compatibility. Moreover, the films with sodium dodecylsulfate were non-irritant for skin, which was confirmed by their in-vitro exposure to the 3D-reconstructed human tissue model.

Effect of process parameters and methylcellulose supplementation on the properties of n-undecane emulsions

ABSTRACT Study was made on the influence of processing parameters on droplet size during emulsification with an Ultra-Turrax homogenizer. Emulsions of undecane in water, stabilized with nonionic Igepals, were prepared at the optimum hydrophilic-lipophilic balance (HLB) of 11.5 and oil-to-water (O/W) ratio of 5/95; the sizes of emulsion droplets were measured at intervals from the commencement of emulsification. Results showed that the optimum emulsification conditions, as pertaining to minimum particle size, arose at 13 400 rpm and 20 minutes of Ultra-Turrax treatment. Additionally, effects exerted by the presence of methylcellulose, time and storage temperature on the emulsions were determined. Emulsions prepared at the optimal processing parameters revealed that the smallest particle sizes (lower than 200 nm) and the best emulsion stability were demonstrated at the oil/water ratios of 3/97 and 5/95, with 3% surfactant content, an HLB value of 10 to 11 and the storage temperature of 25°C, irrespective of the content of methylcellulose. GRAPHICAL ABSTRACT

Adhesion, Proliferation and Migration of NIH/3T3 Cells on Modified Polyaniline Surfaces.

Polyaniline shows great potential and promises wide application in the biomedical field thanks to its intrinsic conductivity and material properties, which closely resemble natural tissues. Surface properties are crucial, as these predetermine any interaction with biological fluids, proteins and cells. An advantage of polyaniline is the simple modification of its surface, e.g., by using various dopant acids. An investigation was made into the adhesion, proliferation and migration of mouse embryonic fibroblasts on pristine polyaniline films and films doped with sulfamic and phosphotungstic acids. In addition, polyaniline films supplemented with poly (2-acrylamido-2-methyl-1-propanesulfonic) acid at various ratios were tested. Results showed that the NIH/3T3 cell line was able to adhere, proliferate and migrate on the pristine polyaniline films as well as those films doped with sulfamic and phosphotungstic acids; thus, utilization of said forms in biomedicine appears promising. Nevertheless, incorporating poly (2-acrylamido-2-methyl-1-propanesulfonic) acid altered the surface properties of the polyaniline films and significantly affected cell behavior. In order to reveal the crucial factor influencing the surface/cell interaction, cell behavior is discussed in the context of the surface energy of individual samples. It was clearly demonstrated that the lesser the difference between the surface energy of the sample and cell, the more cyto-compatible the surface is.