PubMed Health⌕ Search

Biomedical subjects

Eduard Brynda

Publications and source records attributed to Eduard Brynda.

3 recordsLinked to original sources

In vitro hemocompatibility of albumin-heparin multilayer coatings on polyethersulfone prepared by the layer-by-layer technique.

Polyethersulfone foils (PES)--a unique material for blood purification membranes--were coated with a multilayer assembly of heparin (unfractionated or high anticoagulant activity fraction heparin) and albumin (albumin-heparin coatings), or with a multilayer of albumin (albumin coating), using the layer-by-layer technique. The coatings combine advantages of albumin (reduction of nonspecific interactions) and heparin (specific interactions with blood coagulation proteins). The differences between the two heparins, while significant for their biological activity, had only a minor effect on the multilayer assembly with albumin monitored in situ by reflection infrared spectroscopy (FTIR MIRS). Uncoated as well as modified PES surfaces were evaluated using an in vitro assay with freshly drawn, slightly heparinized (1.5 IU heparin/mL) human whole blood. The blood was circulated with a roller pump over the sample surfaces in shear flow across rectangular slit channels ( app. 6 mL/min and 120 s(-1)) for 1.5 h at 37 degrees C. All coatings effectively reduced platelet adhesion and activation according to the PF4 release. The activation of coagulation evaluated as TAT generation was significantly lowered for the coating composed of albumin and high activity heparin. A further beneficial effect of the heparin containing coatings was reduced complement activation as determined by different complement fragments.

Albumins↗

Surface immobilized protein multilayers for cell seeding.

Multilayer assemblies containing various cell-adhesive proteins such as gelatin, collagen IV, and laminin or polycations polylysine and poly(ethyleneimine) were immobilized on the polystyrene surface using the layer-by-layer technique based on hydrophobic and electrostatic interactions between oppositely charged macromolecules. The formation and stability of the assemblies and the adsorption of proteins from a serum containing cell-cultivation media onto their surfaces were observed in real time by Fourier transform infrared multiple internal reflection spectroscopy. The adhesion and growth of mouse embryonic stem cells line D3 were tested in polystyrene culture dishes coated with the assemblies. The cells were seeded in complete serum-containing media or in serum-free media and in the presence (non-differentiated) and/or absence (differentiated) of leukemia inhibitory factor. Proteins from serum-containing cell-cultivation media adsorbed rapidly onto positively charged surfaces. The cells grew best on surfaces coated with gelatin and collagen IV assemblies. There were no significant differences in the growth of the non-differentiated and differentiated cells in complete serum-containing media. When seeded in serum-free media, non-differentiated cells grew better than the differentiated ones. Particularly, polycation surfaces treated with glutaraldehyde promoted the growth of the non-differentiated cells and hindered the growth of the differentiated cells. The layer-by-layer deposition appears to be a practicable technique by which scaffolds for tissue engineering can be coated with biomolecular assemblies tailored to specific cells and applications.

Adsorption↗

The effect of polyelectrolyte chain length on layer-by-layer protein/polyelectrolyte assembly--an experimental study.

The effect of polyelectrolyte chain length on the formation of multilayered assemblies of alternating globular proteins and linear polyanions prepared by the layer-by-layer electrostatic adsorption technique was investigated. The systems studied were albumin/sodium poly(styrenesulfonate), immunoglobulin G/sodium poly(styrenesulfonate), albumin/sodium dextran sulfate, and albumin/heparin. The formation of assemblies was followed using FTIR multiple internal reflection spectroscopy. While the amount of polyelectrolyte adsorbed on the first (primary) protein layer did not depend on its molecular weight, the effect of polyelectrolyte chain length was clearly observed in the following steps of alternating adsorption. Some short-chain polyanion molecules were removed from the surface when a next protein layer was adsorbed from solution. The short polyanion chains were not able to make a sufficient number of ion pairs for stable interaction with additional protein molecules and left the surface as soluble protein/polyanion complexes. The most pronounced effect could be seen with sodium poly(styrenesulfonate) of Mw up to ca. 2 x 10(4), but a detectable effect could be traced even up to Mw ca. 8 x 10(4). Such a pronounced effect, however, was not observed with dextran sulfate. The effect of molecular weight of heparin was clearly observed but all heparins tested, regardless of their molecular weight, effectively assembled with albumin to form multilayer.

Adsorption↗