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    Anomalous influence of salt concentration on deposition of poly(L-lysine)/cellulose sulfate multilayers evidenced by in situ ATR-FTIR
    (Basel : MDPI, 2020) Müller, Martin
    The deposition of polyelectrolyte (PEL) multilayers (PEMs) of poly(L-lysine)/cellulose sulfate (PLL/CS) onto germanium (Ge) substrates depending on salt concentration (cS) and deposition step z at constant PEL concentration cPEL = 0.01 M and pH = 7.0 was studied. In situ ATR-FTIR spectroscopy was used for the quantitative determination of alternate PLL/CS deposition profiles (adsorbed amount versus z) and total deposited PEM amount. By varying cS from 0 M to 1.0 M, a maximum of deposited amount was obtained at 0.1 M, so that both no salinity (0 M) and high salinity (1.0 M) revealed deposited amounts that were far lower than for mean salinity (0.1 M). Furthermore, in situ ATR-FTIR allowed to determine the detailed modulation of the PEL composition during the consecutive PEM deposition, which was interpreted as being due to both diffusion of given PEL from the PEM interior towards the outermost region and release of the PEM upon contact with the bulk oppositely charged PEL solution. Finally, ex situ ATR-FTIR measurements on the PEL solutions after deposition of PEM-20 revealed the distinct release of PEL from the PEM solely for cS = 1.0 M, due to the highest mobility of PEL under high salt conditions. These studies help to prepare functional PEM coatings with defined thicknesses and morphologies for the passivation and activation of material surfaces in the biomedical and food field. © 2020 by the author.
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    Interaction of Poly(l-lysine)/Polysaccharide Complex Nanoparticles with Human Vascular Endothelial Cells
    (Basel : MDPI, 2018) Weber, Dominik; Torger, Bernhard; Richter, Karsten; Nessling, Michelle; Momburg, Frank; Woltmann, Beatrice; Müller, Martin; Schwartz-Albiez, Reinhard
    Angiogenesis plays an important role in both soft and hard tissue regeneration, which can be modulated by therapeutic drugs. If nanoparticles (NP) are used as vectors for drug delivery, they have to encounter endothelial cells (EC) lining the vascular lumen, if applied intravenously. Herein the interaction of unloaded polyelectrolyte complex nanoparticles (PECNP) composed of cationic poly(l-lysine) (PLL) and various anionic polysaccharides with human vascular endothelial cells (HUVEC) was analyzed. In particular PECNP were tested for their cell adhesive properties, their cellular uptake and intracellular localization considering composition and net charge. PECNP may form a platform for both cell coating and drug delivery. PECNP, composed of PLL in combination with the polysaccharides dextran sulfate (DS), cellulose sulfate (CS) or heparin (HEP), either unlabeled or labeled with fluorescein isothiocyanate (FITC) and either with positive or negative net charge were prepared. PECNP were applied to human umbilical cord vein endothelial cells (HUVEC) in both, the volume phase and immobilized phase at model substrates like tissue culture dishes. The attachment of PECNP to the cell surface, their intracellular uptake, and effects on cell proliferation and growth behavior were determined. Immobilized PECNP reduced attachment of HUVEC, most prominently the systems PLL/HEP and PLL/DS. A small percentage of immobilized PECNP was taken up by cells during adhesion. PECNP in the volume phase showed no effect of the net charge sign and only minor effects of the composition on the binding and uptake of PECNP at HUVEC. PECNP were stored in endosomal vesicles in a cumulative manner without apparent further processing. During mitosis, internalized PECNP were almost equally distributed among the dividing cells. Both, in the volume phase and immobilized at the surface, PECNP composed of PLL/HEP and PLL/DS clearly reduced cell proliferation of HUVEC, however without an apparent cytotoxic effect, while PLL/CS composition showed minor impairment. PECNP have an anti-adhesive effect on HUVEC and are taken up by endothelial cells which may negatively influence the proliferation rate of HUVEC. The negative effects were less obvious with the composition PLL/CS. Since uptake and binding for PLL/HEP was more efficient than for PLL/DS, PECNP of PLL/HEP may be used to deliver growth factors to endothelial cells during vascularization of bone reconstitution material, whereas those of PLL/CS may have an advantage for substituting biomimetic bone scaffold material.