04 Fakultät Energie-, Verfahrens- und Biotechnik

Permanent URI for this collectionhttps://elib.uni-stuttgart.de/handle/11682/5

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    Precision 3D‐printed cell scaffolds mimicking native tissue composition and mechanics
    (2020) Erben, Amelie; Hörning, Marcel; Hartmann, Bastian; Becke, Tanja; Eisler, Stephan A.; Southan, Alexander; Cranz, Séverine; Hayden, Oliver; Kneidinger, Nikolaus; Königshoff, Melanie; Lindner, Michael; Tovar, Günter E. M.; Burgstaller, Gerald; Clausen‐Schaumann, Hauke; Sudhop, Stefanie; Heymann, Michael
    Cellular dynamics are modeled by the 3D architecture and mechanics of the extracellular matrix (ECM) and vice versa. These bidirectional cell‐ECM interactions are the basis for all vital tissues, many of which have been investigated in 2D environments over the last decades. Experimental approaches to mimic in vivo cell niches in 3D with the highest biological conformity and resolution can enable new insights into these cell‐ECM interactions including proliferation, differentiation, migration, and invasion assays. Here, two‐photon stereolithography is adopted to print up to mm‐sized high‐precision 3D cell scaffolds at micrometer resolution with defined mechanical properties from protein‐based resins, such as bovine serum albumin or gelatin methacryloyl. By modifying the manufacturing process including two‐pass printing or post‐print crosslinking, high precision scaffolds with varying Young's moduli ranging from 7‐300 kPa are printed and quantified through atomic force microscopy. The impact of varying scaffold topographies on the dynamics of colonizing cells is observed using mouse myoblast cells and a 3D‐lung microtissue replica colonized with primary human lung fibroblast. This approach will allow for a systematic investigation of single‐cell and tissue dynamics in response to defined mechanical and bio‐molecular cues and is ultimately scalable to full organs.
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    Global potentials and costs of synfuels via Fischer-Tropsch process
    (2023) Buchenberg, Patrick; Addanki, Thushara; Franzmann, David; Winkler, Christoph; Lippkau, Felix; Hamacher, Thomas; Kuhn, Philipp; Heinrichs, Heidi; Blesl, Markus
    This paper presents the potentials and costs of synthetic fuels (synfuels) produced by renewable energy via PEM water electrolysis and the subsequent Fischer-Tropsch process for the years 2020, 2030, 2040, and 2050 in selected countries across the globe. The renewable energy potential was determined by the open-source tool pyGRETA and includes photovoltaic, onshore wind, and biomass. Carbon dioxide is obtained from biomass and the atmosphere by direct air capture. The potentials and costs were determined by aggregating minimal cost energy systems for each location on a state level. Each linear energy system was modelled and optimised by the optimisation framework urbs. The analysis focused on decentralised and off-grid synthetic fuels’ production. The transportation costs were roughly estimated based on the distance to the nearest maritime port for export. The distribution infrastructure was not considered since the already-existing infrastructure for fossil fuels can be easily adopted. The results showed that large amounts of synthetic fuels are available for EUR 110/MWh (USD 203/bbl) mainly in Africa, Central and South America, as well as Australia for 2050. This corresponds to a cost reduction of more than half compared to EUR 250/MWh (USD 461/bbl) in 2020. The synfuels’ potentials follow the photovoltaic potentials because of the corresponding low levelised cost of electricity. Batteries are in particular used for photovoltaic-dominant locations, and transportation costs are low compared to production costs.
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    Abschaltung von dezentralen Erzeugungsanlagen infolge von Unterspannungen im Kontext von LFSM-O
    (2022) Schöll, Christian; Lehner, Joachim; Weidner, Johannes; Lens, Hendrik
    Bei einer Überfrequenz jenseits von 50,2 Hz müssen sämtliche Erzeugungsanlagen ihre in das Netz eingespeiste Wirkleistung reduzieren. In den zumeist resistiv geprägten Verteilungsnetzen der Mittel- und Niederspannung führt diese überfrequenzbedingte Leistungsreduktion zu einer lokalen Absenkung des Spannungsprofils. Unterschreiten die Spannungen die Vorgaben aus den Netzanschlussbedingungen, unter denen dezentrale Erzeugungsanlagen die Verbindung zum Netz aufrechterhalten müssen, kann es zu unterspannungsbedingten Schutzabschaltungen kommen. Theoretisch könnten diese bei großflächigem Auftreten zu einer Umkehr des ursprünglichen Überfrequenzereignisses in ein Unterfrequenzereignis führen. Anhand von simulationsbasierten Untersuchungen wird in diesem Beitrag jedoch aufgezeigt, dass unter realitätsnahen Annahmen und unter Berücksichtigung der nationalen Richtlinien sowie der zugehörigen netzplanerischen Leitplanken kein besonderes Risiko hinsichtlich der Anregung der beschriebenen Wirkungskette vorliegt.
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    Identifying and engineering bottlenecks of autotrophic isobutanol formation in recombinant C. ljungdahlii by systemic analysis
    (2021) Hermann, Maria; Teleki, Attila; Weitz, Sandra; Niess, Alexander; Freund, Andreas; Bengelsdorf, Frank Robert; Dürre, Peter; Takors, Ralf
    Clostridium ljungdahlii (C. ljungdahlii, CLJU) is natively endowed producing acetic acid, 2,3-butandiol, and ethanol consuming gas mixtures of CO2, CO, and H2 (syngas). Here, we present the syngas-based isobutanol formation using C. ljungdahlii harboring the recombinant amplification of the “Ehrlich” pathway that converts intracellular KIV to isobutanol. Autotrophic isobutanol production was studied analyzing two different strains in 3-L gassed and stirred bioreactors. Physiological characterization was thoroughly applied together with metabolic profiling and flux balance analysis. Thereof, KIV and pyruvate supply were identified as key “bottlenecking” precursors limiting preliminary isobutanol formation in CLJU[KAIA] to 0.02 g L-1. Additional blocking of valine synthesis in CLJU[KAIA]:ilvE increased isobutanol production by factor 6.5 finally reaching 0.13 g L-1. Future metabolic engineering should focus on debottlenecking NADPH availability, whereas NADH supply is already equilibrated in the current generation of strains.
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    Anti-adhesive surfaces inspired by bee mandible surfaces
    (2023) Saccardi, Leonie; Schiebl, Jonas; Balluff, Franz; Christ, Ulrich; Gorb, Stanislav N.; Kovalev, Alexander; Schwarz, Oliver
    Propolis, a naturally sticky substance used by bees to secure their hives and protect the colony from pathogens, presents a fascinating challenge. Despite its adhesive nature, honeybees adeptly handle propolis with their mandibles. Previous research has shown a combination of an anti-adhesive fluid layer and scale-like microstructures on the inner surface of bee mandibles. Our aim was to deepen our understanding of how surface energy and microstructure influence the reduction in adhesion for challenging substances like propolis. To achieve this, we devised surfaces inspired by the intricate microstructure of bee mandibles, employing diverse techniques including roughening steel surfaces, creating lacquer structures using Bénard cells, and moulding resin surfaces with hexagonal patterns. These approaches generated patterns that mimicked the bee mandible structure to varying degrees. Subsequently, we assessed the adhesion of propolis on these bioinspired structured substrates. Our findings revealed that on rough steel and resin surfaces structured with hexagonal dimples, propolis adhesion was significantly reduced by over 40% compared to unstructured control surfaces. However, in the case of the lacquer surface patterned with Bénard cells, we did not observe a significant reduction in adhesion.
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    Transcriptional CDK inhibitors CYC065 and THZ1 induce apoptosis in glioma stem cells derived from recurrent GBM
    (2021) Juric, Viktorija; Düssmann, Heiko; Lamfers, Martine L. M.; Prehn, Jochen H. M.; Rehm, Markus; Murphy, Brona M.
    Glioma stem cells (GSCs) are tumour initiating cells which contribute to treatment resistance, temozolomide (TMZ) chemotherapy and radiotherapy, in glioblastoma (GBM), the most aggressive adult brain tumour. A major contributor to the uncontrolled tumour cell proliferation in GBM is the hyper activation of cyclin-dependent kinases (CDKs). Due to resistance to standard of care, GBMs relapse in almost all patients. Targeting GSCs using transcriptional CDK inhibitors, CYC065 and THZ1 is a potential novel treatment to prevent relapse of the tumour. TCGA-GBM data analysis has shown that the GSC markers, CD133 and CD44 were significantly upregulated in GBM patient tumours compared to non-tumour tissue. CD133 and CD44 stem cell markers were also expressed in gliomaspheres derived from recurrent GBM tumours. Light Sheet Florescence Microscopy (LSFM) further revealed heterogeneous expression of these GSC markers in gliomaspheres. Gliomaspheres from recurrent tumours were highly sensitive to transcriptional CDK inhibitors, CYC065 and THZ1 and underwent apoptosis while being resistant to TMZ. Apoptotic cell death in GSC subpopulations and non-stem tumour cells resulted in sphere disruption. Collectively, our study highlights the potential of these novel CKIs to induce cell death in GSCs from recurrent tumours, warranting further clinical investigation.
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    Nano-in-micro-particles consisting of PLGA nanoparticles embedded in chitosan microparticles via spray-drying enhances their uptake in the olfactory mucosa
    (2021) Spindler, Lena Marie; Feuerhake, Andreas; Ladel, Simone; Günday, Cemre; Flamm, Johannes; Günday-Türeli, Nazende; Türeli, Emre; Tovar, Günter E. M.; Schindowski, Katharina; Gruber-Traub, Carmen
    Intranasal delivery has gained prominence since 1990, when the olfactory mucosa was recognized as the window to the brain and the central nervous system (CNS); this has enabled the direct site specific targeting of neurological diseases for the first time. Intranasal delivery is a promising route because general limitations, such as the blood-brain barrier (BBB) are circumvented. In the treatment of multiple sclerosis (MS) or Alzheimer’s disease, for example, future treatment prospects include specialized particles as delivery vehicles. Poly(lactic-co-glycolic acid) (PLGA) nanoparticles are well known as promising delivery systems, especially in the area of nose-to-brain (N2B) delivery. Chitosan is also broadly known as a functional additive due to its ability to open tight junctions. In this study, we produced PLGA nanoparticles of different sizes and revealed for the first time their size-time-dependent uptake mechanism into the lamina propria of porcine olfactory mucosa. The intracellular uptake was observed for 80 and 175 nm within only 5 min after application to the epithelium. After 15 min, even 520 nm particles were detected, associated with nuclei. Especially the presence of only 520 nm particles in neuronal fibers is remarkable, implying transcellular and intracellular transport via the olfactory or the trigeminal nerve to the brain and the CNS. Additionally, we developed successfully specialized Nano-in-Micro particles (NiMPs) for the first time via spray drying, consisting of PLGA nanoparticles embedded into chitosan microparticles, characterized by high encapsulation efficiencies up to 51%, reproducible and uniform size distribution, as well as smooth surface. Application of NiMPs accelerated the uptake compared to purely applied PLGA nanoparticles. NiMPs were spread over the whole transverse section of the olfactory mucosa within 15 min. Faster uptake is attributed to additional paracellular transport, which was examined via tight-junction-opening. Furthermore, a separate chitosan penetration gradient of ∼150 µm caused by dissociation from PLGA nanoparticles was observed within 15 min in the lamina propria, which was demonstrated to be proportional to an immunoreactivity gradient of CD14. Due to the beneficial properties of the utilized chitosan-derivative, regarding molecular weight (150-300 kDa), degree of deacetylation (80%), and particle size (0.1-10 µm) we concluded that M2-macrophages herein initiated an anti-inflammatory reaction, which seems to already take place within 15 min following chitosan particle application. In conclusion, we demonstrated the possibility for PLGA nanoparticles, as well as for chitosan NiMPs, to take all three prominent intranasal delivery pathways to the brain and the CNS; namely transcellular, intracellular via neuronal cells, and paracellular transport.
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    Application of ion chromatography for the reliable quantification of ammonium in electrochemical ammonia synthesis experiments : a practical guide
    (2023) Bragulla, Sebastian C. H.; Lorenz, Julian; Harms, Corinna; Wark, Michael; Friedrich, K. Andreas
    Assessing novel electrocatalysts for the electrochemical ammonia synthesis (EAS) requires reliable quantitative trace analysis of electrochemically produced ammonia to infer activity and selectivity. This study concerns the development of an ion chromatography (IC) method for quantitative trace analysis of ammonium in 0.1 M sulfuric acid electrolyte, which is applied to EAS gas-diffusion electrode (GDE) experiments with commercial chromium nitride as electrocatalyst. The developed IC method is highly sensitive, versatile, and reliable, achieving a limit of quantification (LOQ) of 6 μg l-1 (6 ppbmol) ammonium. The impacts of the sample matrix, dilution, and neutralization, as well as contamination, on the quantitative analysis by IC are analyzed. Experimental constraints result in an effective LOQ including dilution of 60 μg l-1 for the determination of ammonium in 0.1 M sulfuric acid electrolyte, owing to necessary sample dilution. The practical guide presented herein is intended to be very relevant for the field of EAS as a guideline and applicable to a broad range of catalyst systems and ion chromatography devices.
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    Light-addressable actuator-sensor platform for monitoring and manipulation of pH gradients in microfluidics : a case study with the enzyme penicillinase
    (2021) Welden, Rene; Jablonski, Melanie; Wege, Christina; Keusgen, Michael; Wagner, Patrick Hermann; Wagner, Torsten; Schöning, Michael J.
    The feasibility of light-addressed detection and manipulation of pH gradients inside an electrochemical microfluidic cell was studied. Local pH changes, induced by a light-addressable electrode (LAE), were detected using a light-addressable potentiometric sensor (LAPS) with different measurement modes representing an actuator-sensor system. Biosensor functionality was examined depending on locally induced pH gradients with the help of the model enzyme penicillinase, which had been immobilized in the microfluidic channel. The surface morphology of the LAE and enzyme-functionalized LAPS was studied by scanning electron microscopy. Furthermore, the penicillin sensitivity of the LAPS inside the microfluidic channel was determined with regard to the analyte’s pH influence on the enzymatic reaction rate. In a final experiment, the LAE-controlled pH inhibition of the enzyme activity was monitored by the LAPS.
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    Lab-scale investigation of palm shell char as tar reforming catalyst
    (2020) Chen, Yen-Hau; Schmid, Max; Chang, Chia-Chi; Chang, Ching-Yuan; Scheffknecht, Günter
    This research investigated the application of palm shell char as a catalyst for the catalytic steam reforming of tar after the sorption enhanced gasification (SEG) process. The catalytic activities of palm shell char and metal-supported palm shell char were tested in a simulated SEG derived syngas with tar model compounds (i.e., toluene and naphthalene) at a concentration of 10 g m-3 NTP. The results indicated that palm shell char had an experimentally excellent catalytic activity for tar reforming with toluene and naphthalene conversions of 0.8 in a short residence time of 0.17 s at 900 °C. A theoretical residence time to reach the complete naphthalene conversion was 1.2 s at 900 °C for palm shell char, demonstrating a promising activity similar to wood char and straw char, but better than CaO. It was also found that potassium and iron-loaded palm shell chars exhibited much better catalytic activity than palm shell char, while the parallel reaction of gasification of K-loaded palm shell char influenced the conversion with its drastic mass loss. Moreover, contrary to CaO, palm shell char presented relatively low selectivity to benzene, and its spontaneous gasification generated extra syngas. In summary, the present study demonstrated that the low-cost material, palm shell char, can successfully be used as the tar-reforming catalyst after SEG process.