Dirk W. Schubert

Spin coating from a molecular point of view: its concentration regimes, influence of molar mass and distribution

Abstract. Spin coating has been investigated utilizing many different polymer solutions. The residual film thickness depends not only on the spinning velocity ω and concentration c0, but also on molar mass M and molar mass distribution – simple models are presented. The concentration dependence is discussed with respect to different effects in low and high concentration regimes. Semi-empirical equations are given for practical use and are compared with experiments. In particular, a model is developed to correlate the Mark-Houwink-Staudinger equation with the residual film thickness which also describes the dependence of film thickness on spinning time consistently with observations by a simple equation. A novel description defining a molecular elementary process during spin coating allows one to describe the influence of molar mass distribution and results in a general definition of a novel spin coat molar mass average, Msp. Spin coating is suggested as a full method for molar mass determination of polymers. Experimental studies are also shown which demonstrate the successful application of the concepts.

Development and characterization of novel electrically conductive PANI-PGS composites for cardiac tissue engineering applications.

Cardiovascular diseases, especially myocardial infarction, are the leading cause of morbidity and mortality in the world, also resulting in huge economic burdens on national economies. A cardiac patch strategy aims at regenerating an infarcted heart by providing healthy functional cells to the injured region via a carrier substrate, and providing mechanical support, thereby preventing deleterious ventricular remodeling. In the present work, polyaniline (PANI) was doped with camphorsulfonic acid and blended with poly(glycerol-sebacate) at ratios of 10, 20 and 30vol.% PANI content to produce electrically conductive composite cardiac patches via the solvent casting method. The composites were characterized in terms of their electrical, mechanical and physicochemical properties. The in vitro biodegradability of the composites was also evaluated. Electrical conductivity increased from 0Scm(-1) for pure PGS to 0.018Scm(-1) for 30vol.% PANI-PGS samples. Moreover, the conductivities were preserved for at least 100h post fabrication. Tensile tests revealed an improvement in the elastic modulus, tensile strength and elasticity with increasing PANI content. The degradation products caused a local drop in pH, which was higher in all composite samples compared with pure PGS, hinting at a buffering effect due to the presence of PANI. Finally, the cytocompatibility of the composites was confirmed when C2C12 cells attached and proliferated on samples with varying PANI content. Furthermore, leaching of acid dopants from the developed composites did not have any deleterious effect on the viability of C2C12 cells. Taken together, these results confirm the potential of PANI-PGS composites for use as substrates to modulate cellular behavior via electrical stimulation, and as biocompatible scaffolds for cardiac tissue engineering applications.

Development and characterization of magnetic iron oxide nanoparticles with a cisplatin-bearing polymer coating for targeted drug delivery

A highly selective and efficient cancer therapy can be achieved using magnetically directed superparamagnetic iron oxide nanoparticles (SPIONs) bearing a sufficient amount of the therapeutic agent. In this project, SPIONs with a dextran and cisplatin-bearing hyaluronic acid coating were successfully synthesized as a novel cisplatin drug delivery system. Transmission electron microscopy images as well as X-ray diffraction analysis showed that the individual magnetite particles were around 4.5 nm in size and monocrystalline. The small crystallite sizes led to the superparamagnetic behavior of the particles, which was exemplified in their magnetization curves, acquired using superconducting quantum interference device measurements. Hyaluronic acid was bound to the initially dextran-coated SPIONs by esterification. The resulting amide bond linkage was verified using Fourier transform infrared spectroscopy. The additional polymer layer increased the vehicle size from 22 nm to 56 nm, with a hyaluronic acid to dextran to magnetite weight ratio of 51:29:20. A maximum payload of 330 μg cisplatin/mL nanoparticle suspension was achieved, thus the particle size was further increased to around 77 nm with a zeta potential of −45 mV. No signs of particle precipitation were observed over a period of at least 8 weeks. Analysis of drug-release kinetics using the dialysis tube method revealed that these were driven by inverse ligand substitution and diffusion through the polymer shell as well as enzymatic degradation of hyaluronic acid. The biological activity of the particles was investigated in a nonadherent Jurkat cell line using flow cytometry. Further, cell viability and proliferation was examined in an adherent PC-3 cell line using xCELLigence analysis. Both tests demonstrated that particles without cisplatin were biocompatible with these cells, whereas particles with the drug induced apoptosis in a dose-dependent manner, with secondary necrosis after prolonged incubation. In conclusion, combination of dextran-coated SPIONs with hyaluronic acid and cisplatin represents a promising approach for magnetic drug targeting in the treatment of cancer.

Evaluation of Fibroblasts Adhesion and Proliferation on Alginate-Gelatin Crosslinked Hydrogel

Due to the relatively poor cell-material interaction of alginate hydrogel, alginate-gelatin crosslinked (ADA-GEL) hydrogel was synthesized through covalent crosslinking of alginate di-aldehyde (ADA) with gelatin that supported cell attachment, spreading and proliferation. This study highlights the evaluation of the physico-chemical properties of synthesized ADA-GEL hydrogels of different compositions compared to alginate in the form of films. Moreover, in vitro cell-material interaction on ADA-GEL hydrogels of different compositions compared to alginate was investigated by using normal human dermal fibroblasts. Viability, attachment, spreading and proliferation of fibroblasts were significantly increased on ADA-GEL hydrogels compared to alginate. Moreover, in vitro cytocompatibility of ADA-GEL hydrogels was found to be increased with increasing gelatin content. These findings indicate that ADA-GEL hydrogel is a promising material for the biomedical applications in tissue-engineering and regeneration.

Toughening and functionalization of bioactive ceramic and glass bone scaffolds by biopolymer coatings and infiltration: a review of the last 5 years

Inorganic scaffolds with high interconnected porosity based on bioactive glasses and ceramics are prime candidates for applications in bone tissue engineering. These materials however exhibit relatively low fracture strength and high brittleness. A simple and effective approach to improve the toughness is to combine the basic scaffold structure with polymer coatings or through the formation of interpenetrating polymer-bioactive ceramic microstructures. The polymeric phase can additionally serve as a carrier for growth factors and therapeutic drugs, thus adding biological functionalities. The present paper reviews the state-of-the art in the field of polymer coated and infiltrated bioactive inorganic scaffolds. Based on the notable combination of bioactivity, improved mechanical properties and drug or growth factor delivery capability, this scaffold type is a candidate for bone and osteochondral regeneration strategies. Remaining challenges for the improvement of the materials are discussed and opportunities to broaden the application potential of this scaffold type are also highlighted.

Biomimetic poly(glycerol sebacate) (PGS) membranes for cardiac patch application.

In this study biomimetic poly(glycerol sebacate) PGS matrix was developed for cardiac patch application. The rationale was that such matrices would provide conducive environment for the seeded cells at the interphase with PGS. From the microstructural standpoint, PGS was fabricated into dense films and porous PGS scaffolds. From the biological aspect, biomimetic PGS membranes were developed via covalently binding peptides Tyr-Ile-Gly-Ser-Arg (YIGSR) and Gly-Arg-Gly-Asp-Ser-Pro (GRGDSP), corresponding to the epitope sequences of laminin and fibronectin, respectively onto the surface. To improve and enhance homogenous binding of peptides onto the PGS surface, chemical modification of its surface was carried out. A sequential regime of alkaline hydrolysis with 0.01 M NaOH for 5 min and acidification with 0.01 M HCl for 25s was optimal. More COOH chemical group was exposed without causing deleterious effect on the bulk properties of the polymer as revealed by the physicochemical analysis carried out. HPLC analysis, chemical imaging and ToF-SIMS were able to establish the successful homogenous functionalization of PGS membranes with the peptides. Finally, the developed biomimetic membranes supported the adhesion and growth of rat and human cardiac progenitor cells.

Preparation and characterization of PHBV microsphere/45S5 bioactive glass composite scaffolds with vancomycin releasing function

PHBV microsphere/45S5 bioactive glass (BG) composite scaffolds with drug release function were developed for bone tissue engineering. BG-based glass-ceramic scaffolds with high porosity (94%) and interconnected pore structure prepared by foam replication method were coated with PHBV microspheres (nominal diameter=3.5 μm) produced by water-in-oil-in-water double emulsion solvent evaporation method. A homogeneous microsphere coating throughout the porous structure of scaffolds was obtained by a simple dip coating method, using the slurry of PHBV microspheres in hexane. Compressive strength tests showed that the microsphere coating slightly improved the mechanical properties of the scaffolds. It was confirmed that the microsphere coating did not inhibit the bioactivity of the scaffolds in SBF. Hydroxyapatite crystals homogeneously grew not only on the struts of the scaffolds but also on the surface of microspheres within 7 days of immersion in SBF. Vancomycin was successfully encapsulated into the PHBV microspheres. The encapsulated vancomycin was released with a dual release profile involving a relatively low initial burst release (21%) and a sustained release (1 month), which is favorable compared to the high initial burst release (77%) and short release period (4 days) measured on uncoated scaffolds. The developed bioactive composite scaffold with drug delivery function has thus the potential to be used advantageously in bone tissue engineering.

Gamma Interferon-Induced Guanylate Binding Protein 1 Is a Novel Actin Cytoskeleton Remodeling Factor

ABSTRACT Gamma interferon (IFN-γ) regulates immune defenses against viruses, intracellular pathogens, and tumors by modulating cell proliferation, migration, invasion, and vesicle trafficking processes. The large GTPase guanylate binding protein 1 (GBP-1) is among the cellular proteins that is the most abundantly induced by IFN-γ and mediates its cell biologic effects. As yet, the molecular mechanisms of action of GBP-1 remain unknown. Applying an interaction proteomics approach, we identified actin as a strong and specific binding partner of GBP-1. Furthermore, GBP-1 colocalized with actin at the subcellular level and was both necessary and sufficient for the extensive remodeling of the fibrous actin structure observed in IFN-γ-exposed cells. These effects were dependent on the oligomerization and the GTPase activity of GBP-1. Purified GBP-1 and actin bound to each other, and this interaction was sufficient to impair the formation of actin filaments in vitro, as demonstrated by atomic force microscopy, dynamic light scattering, and fluorescence-monitored polymerization. Cosedimentation and band shift analyses demonstrated that GBP-1 binds robustly to globular actin and slightly to filamentous actin. This indicated that GBP-1 may induce actin remodeling via globular actin sequestering and/or filament capping. These results establish GBP-1 as a novel member within the family of actin-remodeling proteins specifically mediating IFN-γ-dependent defense strategies.

Electrophoretic deposition of nanostructured-TiO2/chitosan composite coatings on stainless steel

Novel chitosan composite coatings containing titania nanoparticles (n-TiO2) for biomedical applications were developed by electrophoretic deposition (EPD) from ethanol–water suspensions. The optimal ethanol–water ratio was studied in order to avoid bubble formation during the EPD process and to ensure homogeneous coatings. Different n-TiO2 contents (0.5–10 g L−1) were studied for a fixed chitosan concentration (0.5 g L−1) and the properties of the electrophoretic coatings obtained were characterized. Coating composition was analyzed by thermogravimetric analysis (TG), Fourier transform infrared spectroscopy (FTIR) and X-ray diffraction (XRD) analysis. Scanning electron microscopy (SEM) was employed to study both the surface and the cross section morphology of the coatings, and the thicknesses (2–6 μm) of the obtained coatings were correlated with the initial ceramic content. Contact angle measurements, as a preliminary study to predict hypothetic protein attachment on the coatings, were performed for different samples and the influence of a second chitosan layer on top of the coatings was also tested. Finally, the electrochemical behavior of the coatings, evaluated by polarization curves in DMEM at 37 °C, was studied in order to assess the corrosion resistance provided by the n-TiO2/chitosan coatings.

Hybrid hydrogels based on keratin and alginate for tissue engineering

Novel hybrid hydrogels based on alginate and keratin were successfully produced for the first time. The self-assembly properties of keratin, and its ability to mimic the extracellular matrix were combined with the excellent chemical and mechanical stability and biocompatibility of alginate to produce 2D and 3D hybrid hydrogels. These hybrid hydrogels were prepared using two different approaches: sonication, to obtain 2D hydrogels, and a pressure-driven extrusion technique to produce 3D hydrogels. All results indicated that the composition of the hydrogels had a significant effect on their physical properties, and that they can easily be tuned to obtain materials suitable for biological applications. The cell-material interaction was assessed through the use of human umbilical vein endothelial cells, and the results demonstrated that the alginate/keratin hybrid biomaterials supported cell attachment, spreading and proliferation. The results proved that such novel hybrid hydrogels might find applications as scaffolds for soft tissue regeneration.