@article{KruegerGengeBrauneWalteretal.2018, author = {Kr{\"u}ger-Genge, A. and Braune, S. and Walter, M. and Krengel, M. and Kratz, K. and K{\"u}pper, J. H. and Lendlein, Andreas and Jung, Friedrich}, title = {Influence of different surface treatments of poly(n-butyl acrylate) networks on fibroblasts adhesion, morphology and viability}, series = {Clinical hemorheology and microcirculation : blood flow and vessels}, volume = {69}, journal = {Clinical hemorheology and microcirculation : blood flow and vessels}, number = {1-2}, publisher = {IOS Press}, address = {Amsterdam}, issn = {1386-0291}, doi = {10.3233/CH-189130}, pages = {305 -- 316}, year = {2018}, abstract = {BACKGROUND: Physical and chemical characteristics of implant materials determine the fate of long-term cardiovascular devices. However, there is still a lack of fundamental understanding of the molecular mechanisms occurring in the material-tissue interphase. In a previous study, soft covalently crosslinked poly(n-butyl acrylate) networks (cPnBA) were introduced as sterilizable, non-toxic and immuno-compatible biomaterials with mechanical properties adjustable to blood vessels. Here we study the influence of different surface treatments in particular oxygen plasma modification and fibrinogen deposition as well as a combinatorial approach on the adhesion and viability of fibroblasts. RESULTS: Compared to non-treated cPnBAs the advancing water-contact angles were found to be reduced after all surface modifications (p<0.05, each), while lowest values were observed after the combined surface treatment (OPT+FIB). The latter differed significantly from the single OPT and FIB. The number of adherent fibroblasts and their adherence behavior differed on both pristine cPnBA networks. The fibroblast density on cPnBA04 was 743 +/- 434 cells. mm(-2), was about 6.5 times higher than on cPnBA73 with 115 +/- 73 cells. mm(-2). On cPnBA04 about 20\% of the cells were visible as very small, round and buckled cells while all other cells were in a migrating status. On cPnBA73, nearly 50\% of fibroblasts were visible as very small, round and buckled cells. The surface functionalization either using oxygen plasma treatment or fibrinogen coating led to a significant increase of adherent fibroblasts, particularly the combination of both techniques, for both cPnBA networks. It is noteworthy to mention that the fibrinogen coating overruled the characteristics of the pristine surfaces; here, the fibroblast densities after seeding were identical for both cPnBAnetworks. Thus, the binding rather depended on the fibrinogen coating than on the substrate characteristics anymore. While the integrity of the fibroblasts membrane was comparable for both polymers, the MTS tests showed a decreased metabolic activity of the fibroblasts on cPnBA. CONCLUSION: The applied surface treatments of cPnBA successfully improved the adhesion of viable fibroblasts. Under resting conditions as well as after shearing the highest fibroblast densities were found on surfaces with combined post-treatment.}, language = {en} } @article{NieWangXuetal.2019, author = {Nie, Yan and Wang, Weiwei and Xu, Xun and Zou, Jie and Bhuvanesh, Thanga and Schulz, Burkhard and Ma, Nan and Lendlein, Andreas}, title = {Enhancement of human induced pluripotent stem cells adhesion through multilayer laminin coating}, series = {Clinical hemorheology and microcirculation : blood flow and vessels}, volume = {70}, journal = {Clinical hemorheology and microcirculation : blood flow and vessels}, number = {4}, publisher = {IOS Press}, address = {Amsterdam}, issn = {1386-0291}, doi = {10.3233/CH-189318}, pages = {531 -- 542}, year = {2019}, abstract = {Bioengineered cell substrates are a highly promising tool to govern the differentiation of stem cells in vitro and to modulate the cellular behavior in vivo. While this technology works fine for adult stem cells, the cultivation of human induced pluripotent stem cells (hiPSCs) is challenging as these cells typically show poor attachment on the bioengineered substrates, which among other effects causes substantial cell death. Thus, very limited types of surfaces have been demonstrated suitable for hiPSC cultures. The multilayer coating approach that renders the surface with diverse chemical compositions, architectures, and functions can be used to improve the adhesion of hiPSCs on the bioengineered substrates. We hypothesized that a multilayer formation based on the attraction of molecules with opposite charges could functionalize the polystyrene (PS) substrates to improve the adhesion of hiPSCs. Polymeric substrates were stepwise coated, first with dopamine to form a polydopamine (PDA) layer, second with polylysine and last with Laminin-521. The multilayer formation resulted in the variation of hydrophilicity and chemical functionality of the surfaces. Hydrophilicity was detected using captive bubble method and the amount of primary and secondary amines on the surface was quantified by fluorescent staining. The PDA layer effectively immobilized the upper layers and thereby improved the attachment of hiPSCs. Cell adhesion was enhanced on the surfaces coated with multilayers, as compared to those without PDA and/or polylysine. Moreover, hiPSCs spread well over this multilayer laminin substrate. These cells maintained their proliferation capacity and differentiation potential. The multilayer coating strategy is a promising attempt for engineering polymer-based substrates for the cultivation of hiPSCs and of interest for expanding the application scope of hiPSCs.}, language = {en} } @article{HommesSchattmannNeffeAhmadetal.2017, author = {Hommes-Schattmann, Paul J. and Neffe, Axel T. and Ahmad, Bilal and Williams, Gareth R. and Vanneaux, Valerie and Menasche, Philippe and Kalfa, David and Lendlein, Andreas}, title = {RGD constructs with physical anchor groups as polymer co-electrospinnable cell adhesives}, series = {Polymers for advanced technologies}, volume = {28}, journal = {Polymers for advanced technologies}, publisher = {Wiley}, address = {Hoboken}, issn = {1042-7147}, doi = {10.1002/pat.3963}, pages = {1312 -- 1317}, year = {2017}, abstract = {The tissue integration of synthetic polymers can be promoted by displaying RGD peptides at the biointerface with the objective of enhancing colonization of the material by endogenous cells. A firm but flexible attachment of the peptide to the polymer matrix, still allowing interaction with receptors, is therefore of interest. Here, the covalent coupling of flexible physical anchor groups, allowing for temporary immobilization on polymeric surfaces via hydrophobic or dipole-dipole interactions, to a RGD peptide was investigated. For this purpose, a stearate or an oligo(ethylene glycol) (OEG) was attached to GRGDS in 51-69\% yield. The obtained RGD linker constructs were characterized by NMR, IR and MALDI-ToF mass spectrometry, revealing that the commercially available OEG and stearate linkers are in fact mixtures of similar compounds. The RGD linker constructs were co-electrospun with poly(p-dioxanone) (PPDO). After electrospinning, nitrogen could be detected on the surface of the PPDO fibers by X-ray photoelectron spectroscopy. The nitrogen content exceeded the calculated value for the homogeneous material mixture suggesting a pronounced presentation of the peptide on the fiber surface. Increasing amounts of RGD linker constructs in the electrospinning solution did not lead to a detection of an increased amount of peptide on the scaffold surface, suggesting inhomogeneous distribution of the peptide on the PPDO fiber surface. Human adipose-derived stem cells cultured on the patches showed similar viability as when cultured on PPDO containing pristine RGD. The fully characterized RGD linker constructs could serve as valuable tools for the further development of tissue-integrating polymeric scaffolds. Copyright (c) 2016 John Wiley \& Sons, Ltd.}, language = {en} } @article{ProkopovicVikulinaSustretal.2016, author = {Prokopovic, Vladimir Z. and Vikulina, Anna S. and Sustr, David and Duschl, Claus and Volodkin, Dmitry}, title = {Biodegradation-Resistant Multilayers Coated with Gold Nanoparticles. Toward a Tailor-made Artificial Extracellular Matrix}, series = {Journal of colloid and interface science}, volume = {8}, journal = {Journal of colloid and interface science}, publisher = {American Chemical Society}, address = {Washington}, issn = {1944-8244}, doi = {10.1021/acsami.6b10095}, pages = {24345 -- 24349}, year = {2016}, abstract = {Polymer multicomponent coatings such as multilayers mimic an extracellular, matrix (ECM) that attracts significant attention for the use of the multilayers as functional supports for advanced cell culture and tissue engineering. Herein, biodegradation and molecular transport in hyaluronan/polylysine multilayers coated with gold nanoparticles were described. Nanoparticle coating acts as a semipermeable barrier that governs molecular transport into/from the multilayers, and makes them biodegradation-resistant. Model protein lysozyme (mimics of ECM-soluble signals) diffuses into the multilayers as fast- and, slow-diffusing populations existing in an equilibrium,. Such a. composite system may have high potential to be exploited as degradation-resistant drug-delivery platforms suitable for cell-based applications.}, language = {en} } @phdthesis{Uhlig2010, author = {Uhlig, Katja}, title = {Untersuchungen PEG-basierter thermo-responsiver Polymeroberfl{\"a}chen zur Steuerung der Zelladh{\"a}sion}, url = {http://nbn-resolving.de/urn:nbn:de:kobv:517-opus-47784}, school = {Universit{\"a}t Potsdam}, year = {2010}, abstract = {Moderne Methoden f{\"u}r die Einzelzellanalyse werden dank der fortschreitenden Weiterentwicklung immer sensitiver. Dabei steigen jedoch auch die Anforderungen an das Probenmaterial. Viele Aufbereitungsprotokolle adh{\"a}renter Zellen beinhalten eine enzymatische Spaltung der Oberfl{\"a}chenproteine, um die Abl{\"o}sung vom Zellkultursubstrat zu erm{\"o}glichen. Verschiedene Methoden, wie die Patch-Clamp-Technik oder eine auf der Markierung extrazellul{\"a}rer Dom{\"a}nen von Membranproteinen basierende Durchflusszytometrie k{\"o}nnen dann nur noch eingeschr{\"a}nkt eingesetzt werden. Daher ist die Etablierung neuer Zellabl{\"o}semethoden dringend notwendig. In der vorliegenden Arbeit werden erstmals PEG-basierte thermo-responsive Oberfl{\"a}chen erfolgreich f{\"u}r die Zellkultur eingesetzt. Dabei wird das zerst{\"o}rungsfreie Abl{\"o}sen verschiedener Zelllinien von den Oberfl{\"a}chen durch Temperatursenkung realisiert. Die Funktionalit{\"a}t der Oberfl{\"a}chen wird durch Variation der Polymerstruktur, sowie der Konzentration der Beschichtungsl{\"o}sung, durch Beschichtung der Oberfl{\"a}chen mit einem zelladh{\"a}sionsf{\"o}rdernden Protein (Fibronektin) und durch Adsorption zelladh{\"a}sionsvermittelnder Peptide (RGD) optimiert. Um den Zellabl{\"o}sungsprozess detaillierter zu untersuchen, wird hier zum ersten Mal der direkte Zellkontakt mit thermo-responsiven Oberfl{\"a}chen mittels oberfl{\"a}chensensitiver Mikroskopie (TIRAF) sichtbar gemacht. Mit dieser Technik sind die exakte Quantifizierung und die Analyse der Reduktion der Zelladh{\"a}sionsfl{\"a}che w{\"a}hrend des Abk{\"u}hlens m{\"o}glich. Hierbei werden in Abh{\"a}ngigkeit von der Zelllinie Unterschiede im Zellverhalten w{\"a}hrend des Abl{\"o}sens festgestellt: Zellen, wie eine Brustkrebszelllinie und eine Ovarzelllinie, die bekanntermaßen st{\"a}rker mit ihrer Umgebung in Kontakt treten, vergr{\"o}ßern im Verlauf des Beobachtungszeitraumes den Abstand zwischen Zellmembran und Oberfl{\"a}che, reduzieren jedoch ihre Zell-Substratkontaktfl{\"a}che kaum. Mausfibroblasten hingegen verkleinern drastisch die Zelladh{\"a}sionsfl{\"a}che. Der Abl{\"o}sungsprozess wird vermutlich aktiv von den Zellen gesteuert. Diese Annahme wird durch zwei Beobachtungen gest{\"u}tzt: Erstens verl{\"a}uft die Reduktion der Zelladh{\"a}sionsfl{\"a}che bei Einschr{\"a}nkung des Zellmetabolismus durch eine Temperatursenkung auf 4 °C verz{\"o}gert. Zweitens hinterlassen die Zellen Spuren, die nach dem Abl{\"o}sen der Zellen auf den Oberfl{\"a}chen zur{\"u}ckbleiben. Mittels Kombination von TIRAF- und TIRF-Mikroskopie werden die Zelladh{\"a}sionsfl{\"a}che und die Aktinstruktur gleichzeitig beobachtet. Die Verkn{\"u}pfung beider Methoden stellt eine neue M{\"o}glichkeit dar, intrazellul{\"a}re Prozesse mit der Zellabl{\"o}sung von thermo-responsiven Oberfl{\"a}chen zu korrelieren.}, language = {de} } @phdthesis{Korn2007, author = {Korn, Christian}, title = {Stochastic dynamics of cell adhesion in hydrodynamic flow}, url = {http://nbn-resolving.de/urn:nbn:de:kobv:517-opus-12997}, school = {Universit{\"a}t Potsdam}, year = {2007}, abstract = {In this thesis the interplay between hydrodynamic transport and specific adhesion is theoretically investigated. An important biological motivation for this work is the rolling adhesion of white blood cells experimentally investigated in flow chambers. There, specific adhesion is mediated by weak bonds between complementary molecular building blocks which are either located on the cell surface (receptors) or attached to the bottom plate of the flow chamber (ligands). The model system under consideration is a hard sphere covered with receptors moving above a planar ligand-bearing wall. The motion of the sphere is influenced by a simple shear flow, deterministic forces, and Brownian motion. An algorithm is given that allows to numerically simulate this motion as well as the formation and rupture of bonds between receptors and ligands. The presented algorithm spatially resolves receptors and ligands. This opens up the perspective to apply the results also to flow chamber experiments done with patterned substrates based on modern nanotechnological developments. In the first part the influence of flow rate, as well as of the number and geometry of receptors and ligands, on the probability for initial binding is studied. This is done by determining the mean time that elapses until the first encounter between a receptor and a ligand occurs. It turns out that besides the number of receptors, especially the height by which the receptors are elevated above the surface of the sphere plays an important role. These findings are in good agreement with observations of actual biological systems like white blood cells or malaria-infected red blood cells. Then, the influence of bonds which have formed between receptors and ligands, but easily rupture in response to force, on the motion of the sphere is studied. It is demonstrated that different states of motion-for example rolling-can be distinguished. The appearance of these states depending on important model parameters is then systematically investigated. Furthermore, it is shown by which bond property the ability of cells to stably roll in a large range of applied flow rates is increased. Finally, the model is applied to another biological process, the transport of spherical cargo particles by molecular motors. In analogy to the so far described systems molecular motors can be considered as bonds that are able to actively move. In this part of the thesis the mean distance the cargo particles are transported is determined.}, language = {en} } @phdthesis{Erdmann2005, author = {Erdmann, Thorsten}, title = {Stochastic dynamics of adhesion clusters under force}, url = {http://nbn-resolving.de/urn:nbn:de:kobv:517-opus-5564}, school = {Universit{\"a}t Potsdam}, year = {2005}, abstract = {Adhesion of biological cells to their environment is mediated by two-dimensional clusters of specific adhesion molecules which are assembled in the plasma membrane of the cells. Due to the activity of the cells or external influences, these adhesion sites are usually subject to physical forces. In recent years, the influence of such forces on the stability of cellular adhesion clusters was increasingly investigated. In particular, experimental methods that were originally designed for the investigation of single bond rupture under force have been applied to investigate the rupture of adhesion clusters. The transition from single to multiple bonds, however, is not trivial and requires theoretical modelling. Rupture of biological adhesion molecules is a thermally activated, stochastic process. In this work, a stochastic model for the rupture and rebinding dynamics of clusters of parallel adhesion molecules under force is presented. In particular, the influence of (i) a constant force as it may be assumed for cellular adhesion clusters is investigated and (ii) the influence of a linearly increasing force as commonly used in experiments is considered. Special attention is paid to the force-mediated cooperativity of parallel adhesion bonds. Finally, the influence of a finite distance between receptors and ligands on the binding dynamics is investigated. Thereby, the distance can be bridged by polymeric linker molecules which tether the ligands to a substrate.}, subject = {Biophysik}, language = {en} } @phdthesis{Schwarz2004, author = {Schwarz, Ulrich Sebastian}, title = {Forces and elasticity in cell adhesion}, url = {http://nbn-resolving.de/urn:nbn:de:kobv:517-0001343}, school = {Universit{\"a}t Potsdam}, year = {2004}, abstract = {Das Verhalten adh{\"a}renter Zellen h{\"a}ngt stark von den chemischen, topographischen und mechanischen Eigenschaften ihrer Umgebung ab. Experimentelle Untersuchungen der letzten Jahre haben gezeigt, dass adh{\"a}rente Zellen aktiv die elastischen Eigenschaften ihrer Umgebung erkunden, indem sie an dieser ziehen. Der resultierende Kraftaufbau h{\"a}ngt von den elastischen Eigenschaften der Umgebung ab und wird an den Adh{\"a}sionskontakten in entsprechende biochemische Signale umgewandelt, die zellul{\"a}re Programme wie Wachstum, Differenzierung, programmierten Zelltod und Zellbewegung mitbestimmen. Im Allgemeinen sind Kr{\"a}fte wichtige Einflussgr{\"o}ßen in biologischen Systemen. Weitere Beispiele daf{\"u}r sind H{\"o}r- und Tastsinn, Wundheilung sowie die rollende Adh{\"a}sion von weißen Blutk{\"o}rperchen auf den W{\"a}nden der Blutgef{\"a}ße. In der Habilitationsschrift von Ulrich Schwarz werden mehrere theoretische Projekte vorgestellt, die die Rolle von Kr{\"a}ften und Elastizit{\"a}t in der Zelladh{\"a}sion untersuchen. (1) Es wurde eine neue Methode entwickelt, um die Kr{\"a}fte auszurechnen, die Zellen an den Kontaktpunkten auf mikro-strukturierte elastische Substrate aus{\"u}ben. Das Hauptergebnis ist, dass Zell-Matrix-Kontakte als Mechanosensoren funktionieren, an denen interne Kr{\"a}fte in Proteinaggregation umgewandelt werden. (2) Eine Ein-Schritt-Master-Gleichung, die die stochastische Dynamik von Adh{\"a}sionsclustern als Funktion von Clustergr{\"o}ße, R{\"u}ckbindungsrate und Kraft beschreibt, wurde sowohl analytisch als auch numerisch gel{\"o}st. Zudem wurde dieses Modell auf Zell-Matrix-Kontakte, dynamische Kraftspektroskopie sowie die rollende Adh{\"a}sion angewandt. (3) Im Rahmen der linearen Elastizit{\"a}tstheorie und mit Hilfe des Konzepts der Kraftdipole wurde ein Modell formuliert und gel{\"o}st, das die Positionierung und Orientierung von Zellen in weicher Umgebung vorhersagt. Diese Vorhersagen sind in guter {\"U}bereinstimmung mit zahlreichen experimentellen Beobachtungen f{\"u}r Fibroblasten auf elastischen Substraten und in Kollagen-Gelen.}, language = {en} } @phdthesis{Groth2003, author = {Groth, Thomas}, title = {Die Bedeutung der Volumen- und Oberfl{\"a}cheneigenschaften von Biomaterialien f{\"u}r die Adsorption von Proteinen und nachfolgende zellul{\"a}re Reaktionen}, url = {http://nbn-resolving.de/urn:nbn:de:kobv:517-0001022}, school = {Universit{\"a}t Potsdam}, year = {2003}, abstract = {Es ist schon seit l{\"a}ngerer Zeit bekannt, dass nach Kontakt des Biomaterials mit der biologischen Umgebung bei Implantation oder extrakorporaler Wechselwirkung zun{\"a}chst Proteine aus dem umgebenden Milieu adsorbiert werden, wobei die Oberfl{\"a}cheneigenschaften des Materials die Zusammensetzung der Proteinschicht und die Konformation der darin enthaltenden Proteine determinieren. Die nachfolgende Wechselwirkung von Zellen mit dem Material wird deshalb i.d.R. von der Adsorbatschicht vermittelt. Der Einfluss der Oberfl{\"a}chen auf die Zusammensetzung und Konformation der Proteine und die nachfolgende Wechselwirkung mit Zellen ist von besonderem Interesse, da einerseits eine Aussage {\"u}ber die Anwendbarkeit erm{\"o}glicht wird, andererseits Erkenntnisse {\"u}ber diese Zusammenh{\"a}nge f{\"u}r die Entwicklung neuer Materialien mit verbesserter Biokompatibilit{\"a}t genutzt werden k{\"o}nnen. In der vorliegenden Habilitationsschrift wurde deshalb der Einfluss der Zusammensetzung von Polymeren bzw. von deren Oberfl{\"a}cheneigenschaften auf die Adsorption von Proteinen, den Aktivit{\"a}tszustand der plasmatischen Gerinnung und die Adh{\"a}sion von Zellen untersucht. Dabei wurden auch M{\"o}glichkeiten zur Beeinflussung dieser Vorg{\"a}nge {\"u}ber eine Ver{\"a}nderung der Volumenzusammensetzung oder durch Oberfl{\"a}chenmodifikationen von Biomaterialien vorgestellt. Erkenntnisse aus diesen Arbeiten konnten f{\"u}r die Entwicklung von Membranen f{\"u}r Biohybrid-Organe genutzt werden.}, language = {de} }