Zdenka Kuceková

Colloidal polyaniline dispersions: Antibacterial activity, cytotoxicity and neutrophil oxidative burst

Polyaniline colloids rank among promising application forms of this conducting polymer. Cytotoxicity, antibacterial activity, and neutrophil oxidative burst tests were performed on cells treated with colloidal polyaniline dispersions. The antibacterial effect of colloidal polyaniline against gram-positive and gram-negative bacteria was most pronounced for Bacillus cereus and Escherichia coli, with a minimum inhibitory concentration of 3,500 μg mL(-1). The data recorded on human keratinocyte (HaCaT) and a mouse embryonic fibroblast (NIH/3T3) cell lines using an MTT assay and flow cytometry indicated a concentration-dependent cytotoxicity of colloid, with the absence of cytotoxic effect at around 150 μg mL(-1). The neutrophil oxidative burst test then showed that colloidal polyaniline, in concentrations <150 μg mL(-1), was not able to stimulate the production of reactive oxygen species in neutrophils and whole human blood. However, it worked efficiently as a scavenger of those already formed.

Antibacterial properties of polyaniline-silver films

In situ polymerised thin polyaniline (PANI) films produced on polystyrene dishes were tested for their antibacterial activity with respect to Escherichia coli and Staphylococcus aureus, representing both gram-positive and gram-negative bacteria. PANI films were subsequently used for the reduction of silver ions to metallic Ag. PANI salt and base in original forms and after the deposition of Ag were studied. PANI salt showed a significant antibacterial effect against both bacteria strains while the efficacy of neat PANI base was only marginal. After the Ag deposition, the PANI base exhibited different levels of antibacterial effect depending on the type of the bacterial strain; the growth of gram-positive Staphylococcus aureus was inhibited depending on the Ag concentration on the film, while Escherichia coli remained uninfluenced. Efficacy of the PANI salt with deposited Ag against both bacteria strains was comparable with that of PANI alone and was not affected by the Ag concentration. The results show that Ag deposition can be a suitable method for the preparation of PANI base films with improved antibacterial properties.

Phenolic compounds from Allium schoenoprasum, Tragopogon pratensis and Rumex acetosa and their antiproliferative effects.

Experimental studies have shown that phenolic compounds have antiproliferative and tumour arresting effects. The aim of this original study was to investigate the content of phenolic compounds (PhC) in flowers of Allium schoenoprasum (chive), Tragopogon pratensis (meadow salsify) and Rumex acetosa (common sorrel) and their effect on proliferation of HaCaT cells. Antiproliferative effects were evaluated in vitro using the following concentrations of phenolic compounds in cultivation medium: 100, 75, 50 and 25 µg/mL. Phenolic composition was also determined by HPLC. The results indicate that even low concentrations of these flowers’ phenolic compounds inhibited cell proliferation significantly and the possible use of the studied herb’s flowers as sources of active phenolic compounds for human nutrition.

Edible flowers — antioxidant activity and impact on cell viability

The phenolic compound composition, antioxidant activity and impact on cell viability of edible flower extracts of Allium schoenoprasum; Bellis perennis; Cichorium intybus; Rumex acetosa; Salvia pratensis; Sambucus nigra; Taraxacum officinale; Tragopogon pratensis; Trifolium repens and Viola arvensis was examined for the first time. Total phenolic content of the flowers of these plants fell between 11.72 and 42.74 mg of tannin equivalents/kg of dry matter. Antioxidant activity ranged from 35.56 to 71.62 g of ascorbic acid equivalents/kg of dry matter. Using the Human Hepatocellular Carcinoma cell-line (HepG2) and the Human Immortalized Non-tumorigenic Keratinocyte cell line (HaCaT), we assessed cell viability following a 3 day incubation period in media containing 25, 50, 75 and 100 μg/ml of total phenolic compounds using a colorimetric MTT assay. These three properties could make the herbs useful in treatment of various diseases like cancer. The tested extracts had significant effects on cell viability, but the effects were dependent not only on the phenolic compound concentration and the edible flowers species, but also on the phenolic compound and antioxidant profiles. In addition, responses differed between cell lines.

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.

The chemical stability and cytotoxicity of carbonyl iron particles grafted with poly(glycidyl methacrylate) and the magnetorheological activity of their suspensions

Carbonyl iron (CI) particles were grafted with poly(glycidyl methacrylate) (PGMA) using atom transfer radical polymerization. Compact coating of PGMA largely improved the chemical stability of the particles in an acid environment and thus reduced the common drawback of bare CI particles. Furthermore, due to possible medical applications of CI-polymer systems for magnetic drug targeting, an in vitro cytotoxicity test was performed using an NIH/3T3 cell line. The cell viability was evaluated by spectrometric assay (MTT). The results show that the prepared particles are not cytotoxic. Moreover, bare CI particles as well as synthesized core–shell particles were suspended in silicone oil, and the rheological behavior of MR suspensions was investigated in controlled shear rate mode under various magnetic field strengths. Dynamic yield stress as a measure of the rigidity of the created internal structures of the suspensions was determined using the Herschel–Bulkley model, which provided a reasonably good fit for rheological data. MR suspensions of PGMA-coated particles exhibited only slightly decreased yield stresses due to their negligibly-affected magnetic performance.

Polyphenolic extracts of edible flowers incorporated onto atelocollagen matrices and their effect on cell viability.

The phenolic extract of chives flowers (Allium schoenoprasum, Liliaceae), introduced Sage (Salvia pratensis, Lamiaceae), European elderberry (Sambucus nigra, Caprifoliaceae) and common dandelion (Taraxacum officinale, Asteraceae) were characterised by High Performance Liquid Chromatography and incorporated in different concentrations onto atelocollagen thin films. In order to assess the biological impact of these phenolic compounds on cell viability, human immortalised non-tumorigenic keratinocyte cell line was seeded on the thin films and cell proliferation was determined by using an MTT assay. In addition, their antimicrobial activity was estimated by using an agar diffusion test. Data indicated the concomitance between cell viability and concentration of polyphenols. These findings suggest that these phenolic-endowed atelocollagen films might be suitable for tissue engineering applications, on account of the combined activity of polyphenols and collagen.

Properties of biomineralized (CaCO3) PVP-CMC hydrogel with reference to its cytotoxicity

ABSTRACT The authors focus on properties of biomineralized (CaCO3) PVP-CMC hydrogel (designated as I–X) including cytotoxicity assay using primary mouse embryonic fibroblasts. The biomineralized samples (VII–X) showed >80% cell viability, was selected for further characterizations. FTIR and XRD indicate deposition of CaCO3 within the PVP-CMC hydrogel matrix, SEM shows changes in morphology and pore diameter (VII and VIII: 1–12 µm; IX: 10–70 µm; X: 70–170 µm), TGA determines the decomposition scenario of CaCO3, and tensile strength of samples (VII–X) ranged between 0.04 and 1.0 GPa, which practically corresponds to the modulus of cancellous bone. GRAPHICAL ABSTRACT

Developing a biomaterial interface based on poly(lactic acid) via plasma-assisted covalent anchorage of d-glucosamine and its potential for tissue regeneration.

The aim of this study was to develop the potential tissue engineering applications of d-glucosamine (GlcN) immobilized onto the surface of a biodegradable matrix in order to induce a desired biological effect at biointerfaces. Thus, for sample preparation we used a novel multistep physicochemical approach. In the first step the poly(lactic acid) (PLA) films were exposed to a low pressure plasma in air atmosphere, followed by radical graft copolymerization with acrylic acid to yield a carboxyl-functionalized spacer layer on the PLA surface. The carboxyl groups were then coupled to GlcN molecules via the carbodiimide chemistry. The developed surfaces were characterized by X-ray Photoelectron Spectroscopy (XPS), Contact angle measurements and Atomic Force Microscopy (AFM). A preliminary study on the proliferation of fibroblasts on the developed surfaces was performed using the NIH/3T3 cell line.