Publikationen am Institut für Chemie und Biotechnik

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Dorner-Reisel, Annett; Ritter, Uwe; Moje, Jens; Freiberger, Emma; Scharff, Peter
Effect of fullerene C60 thermal and tribomechanical loading on Raman signals. - In: Diamond and related materials, ISSN 0925-9635, Bd. 126 (2022), 109036, S. 1-14

Fullerene C60 powder was loaded by 1 N normal force and exposed to sliding under different frequencies for 15 min. It is shown that the velocity of the sliding movement determines the stability of the fullerene C60 powder. At slow velocity of movement with a frequency of 1 Hz under 1 N normal force, the fullerene C60 structure remains undamaged after 15 min sliding. On the contrary, high sliding velocities of 10 Hz and 50 Hz affected fragmentation of the fullerene C60, which resulted in a reduction of the coefficient of friction (COF). During sliding with 1 Hz, the friction reached the highest level with an average COF of 0.59 ± 0.03. The faster relative motion under 1 N normal force gave a lower average COF with 0.39 ± 0.03. The initial fullerene C60 powder formed a thick compressed layer in the tribomechanical loaded zone. As proven by Raman spectroscopy, operating the tribomechanical sliding test at 50 Hz stimulated the re-attraction of fresh C60 fullerene island onto the fragmented layer from outside of the loaded powder regions. The COF was increasing again up to 0.44 ± 0.04 for 1 N normal force and 50 Hz frequency. The fragmentation and decomposition of fullerene C60 with increasing sliding velocity is attributed to thermal heating up during fast relative movement. Raman spectra of the tribomechanical loaded fullerene C60 are compared with Raman spectra from slowly heated up C60 in air and with Raman spectra of laser irradiated fullerene C60.



https://doi.org/10.1016/j.diamond.2022.109036
Mai, Patrick; Hampl, Jörg; Bača, Martin; Brauer, Dana; Singh, Sukhdeep; Weise, Frank; Borowiec, Justyna; Schmidt, André; Küstner, Johanna Merle; Klett, Maren; Gebinoga, Michael; Schroeder, Insa S.; Markert, Udo R.; Glahn, Felix; Schumann, Berit; Eckstein, Diana; Schober, Andreas
MatriGrid® based biological morphologies: tools for 3D cell culturing. - In: Bioengineering, ISSN 2306-5354, Bd. 9 (2022), 5, 220, S. 1-41

Recent trends in 3D cell culturing has placed organotypic tissue models at another level. Now, not only is the microenvironment at the cynosure of this research, but rather, microscopic geometrical parameters are also decisive for mimicking a tissue model. Over the years, technologies such as micromachining, 3D printing, and hydrogels are making the foundation of this field. However, mimicking the topography of a particular tissue-relevant substrate can be achieved relatively simply with so-called template or morphology transfer techniques. Over the last 15 years, in one such research venture, we have been investigating a micro thermoforming technique as a facile tool for generating bioinspired topographies. We call them MatriGrid®s. In this research account, we summarize our learning outcome from this technique in terms of the influence of 3D micro morphologies on different cell cultures that we have tested in our laboratory. An integral part of this research is the evolution of unavoidable aspects such as possible label-free sensing and fluidic automatization. The development in the research field is also documented in this account.



https://doi.org/10.3390/bioengineering9050220
Bača, Martin; Brauer, Dana; Klett, Maren; Fernekorn, Uta; Singh, Sukhdeep; Hampl, Jörg; Groß, Gregor Alexander; Mai, Patrick; Friedel, Karin; Schober, Andreas
Automated analysis of acetaminophen toxicity on 3D HepaRG cell culture in microbioreactor. - In: Bioengineering, ISSN 2306-5354, Bd. 9 (2022), 5, 196, S. 1-16

Real-time monitoring of bioanalytes in organotypic cell cultivation devices is a major research challenge in establishing stand-alone diagnostic systems. Presently, no general technical facility is available that offers a plug-in system for bioanalytics in diversely available organotypic culture models. Therefore, each analytical device has to be tuned according to the microfluidic and interface environment of the 3D in vitro system. Herein, we report the design and function of a 3D automated culture and analysis device (3D-ACAD) which actively perfuses a custom-made 3D microbioreactor, samples the culture medium and simultaneously performs capillary-based flow ELISA. A microstructured MatriGrid® has been explored as a 3D scaffold for culturing HepaRG cells, with albumin investigated as a bioanalytical marker using flow ELISA. We investigated the effect of acetaminophen (APAP) on the albumin secretion of HepaRG cells over 96 h and compared this with the albumin secretion of 2D monolayer HepaRG cultures. Automated on-line monitoring of albumin secretion in the 3D in vitro mode revealed that the application of hepatotoxic drug-like APAP results in decreased albumin secretion. Furthermore, a higher sensitivity of the HepaRG cell culture in the automated 3D-ACAD system to APAP was observed compared to HepaRG cells cultivated as a monolayer. The results support the use of the 3D-ACAD model as a stand-alone device, working in real time and capable of analyzing the condition of the cell culture by measuring a functional analyte. Information obtained from our system is compared with conventional cell culture and plate ELISA, the results of which are presented herein.



https://doi.org/10.3390/bioengineering9050196
Tong, Ciqing; Wondergem, Joeri A. J.; van den Brink, Marijn; Kwakernaak, Markus C.; Chen, Ying; Hendrix, Marco M. R. M.; Voets, Ilja K.; Danen, Erik Hendrik Julius; Le Dévédec, Sylvia; Heinrich, Doris; Kieltyka, Roxanne E.
Spatial and temporal modulation of cell instructive cues in a filamentous supramolecular biomaterial. - In: ACS applied materials & interfaces, ISSN 1944-8252, Bd. 14 (2022), 15, S. 17042-17054

Supramolecular materials provide unique opportunities to mimic both the structure and mechanics of the biopolymer networks that compose the extracellular matrix. However, strategies to modify their filamentous structures in space and time in 3D cell culture to study cell behavior as encountered in development and disease are lacking. We herein disclose a multicomponent squaramide-based supramolecular material whose mechanics and bioactivity can be controlled by light through co-assembly of a 1,2-dithiolane (DT) monomer that forms disulfide cross-links. Remarkably, increases in storage modulus from ∼200 Pa to >10 kPa after stepwise photo-cross-linking can be realized without an initiator while retaining colorlessness and clarity. Moreover, viscoelasticity and plasticity of the supramolecular networks decrease upon photo-irradiation, reducing cellular protrusion formation and motility when performed at the onset of cell culture. When applied during 3D cell culture, force-mediated manipulation is impeded and cells move primarily along earlier formed channels in the materials. Additionally, we show photopatterning of peptide cues in 3D using either a photomask or direct laser writing. We demonstrate that these squaramide-based filamentous materials can be applied to the development of synthetic and biomimetic 3D in vitro cell and disease models, where their secondary cross-linking enables mechanical heterogeneity and shaping at multiple length scales.



https://doi.org/10.1021/acsami.1c24114
Visaveliya, Nikunjkumar R.; Mazétyté-Stasinskiené, Raminta; Köhler, Michael
Stationary, continuous, and sequential surface-enhanced raman scattering sensing based on the nanoscale and microscale polymer-metal composite sensor particles through microfluidics: a review. - In: Advanced optical materials, ISSN 2195-1071, Bd. 10 (2022), 7, 2102757, S. 1-25

Surface-enhanced Raman scattering (SERS) is a label-free and accurate analytical technique for the detection of a broad range of various analytes such as, biomolecules, pesticides, petrochemicals, as well as, cellular and other biological systems. A key component for the SERS analysis is the substrate which is required to be equipped with plasmonic features of metal nanostructures that directly interact with light and targeted analytes. Either metal nanoparticles can be deposited on the solid support (glass or silicon) which is suitable for stationary SERS analysis or dispersed in the solution (freely moving nanoparticles). Besides these routinely utilizing SERS substrates, polymer-metal composite particles are promising for sustained SERS analysis where metal nanoparticles act as plasmon-active (hence SERS-active) components and polymer particles act as support to the metal nanoparticles. Composite sensor particles provide 3D interaction possibilities for analytes, suitable for stationary, continuous, and sequential analysis, and they are reusable/regenerated. Therefore, this review is focused on the experimental procedures for the development of multiscale, uniform, and reproducible composite sensor particles together with their application for SERS analysis. The microfluidic reaction technique is highly versatile in the production of uniform and size-tunable composite particles, as well as, for conducting SERS analysis.



https://doi.org/10.1002/adom.202102757
Nozdrenko, Dmytro; Matvienko, Tatiana; Vygovska, Oksana; Soroca, Vasil; Bogutska, Kateryna; Zholos, Alexander; Scharff, Peter; Ritter, Uwe; Prylutskyy, Yuriy
Post-traumatic recovery of muscle soleus in rats is improved via synergistic effect of C60 fullerene and TRPM8 agonist menthol. - In: Applied nanoscience, ISSN 2190-5517, Bd. 12 (2022), 3, S. 467-478

Functional biomechanical parameters of muscle soleus contraction in rats as well as selected blood biochemical parameters were studied during the first 3 days of post-traumatic syndrome progression caused by the destruction of muscle cells by compression. Single administration of the antioxidant C60 fullerene and the selective agonist of TRPM8 channels menthol were used as therapeutic agents. Injection of C60 fullerene at a concentration of 1 mg/kg into the damaged muscle improved its contractile function by 25-28%. The use of combined injections of C60 fullerene and menthol (at the concentration 1 mg/kg) improved this index by additional 27-39% and simultaneously stabilized the decrease in muscle strength observed throughout the experiment. A tendency towards a decrease in the indexes of the above described biochemical parameters by 10-15% were found with the therapeutic administration of C60 fullerene. With combined injections of C60 fullerene and menthol, the above described biochemical parameters decreased by an additional 17-24%. The synergism between the action of menthol and C60 fullerene on the post-traumatic recovery of skeletal muscle function opens up new perspectives for the clinical application of this combination therapy.



https://doi.org/10.1007/s13204-021-01703-z
Visaveliya, Nikunjkumar R.; Mazétyté-Stasinskiené, Raminta; Köhler, Michael
General background of SERS sensing and perspectives on polymer-supported plasmon-active multiscale and hierarchical sensor particles. - In: Advanced optical materials, ISSN 2195-1071, Bd. 10 (2022), 4, 2102001, S. 1-27

Surface-enhanced Raman scattering (SERS) is one of the most powerful analytical techniques for the identification of molecules. The substrate, on which SERS is dependent, contains regions of nanoscale gaps (hotspots) that hold the ability to concentrate incident electromagnetic fields and effectively amplify vibrational scattering signals of adsorbed analytes. While surface plasmon resonance from metal nanostructures is a central focus for the SERS effect, the support of polymers can be significantly advantageous to provide larger exposure of structured metal surfaces for efficient interactions with analytes. Characteristics of the polymer particles such as softness, flexibility, swellability, porosity, optical transparency, metal-loading ability, and high surface area can allow diffusion of analytes and penetrating light deeply that can enormously amplify sensing outcomes. As polymer-supported plasmon-active sensor particles can emerge as versatile SERS substrates, the microfluidic platform is promising for the generation of sensor particles as well as for performing sequential SERS analysis of multiple analytes. Therefore, in this perspective article, the development of multifunctional polymer-metal composite particles, and their applications as potential sensors for SERS sensing through microfluidics are presented. A detailed background from the beginning of the SERS field and perspectives for the multifunctional sensor particles for efficient SERS sensing are provided.



https://doi.org/10.1002/adom.202102001
Nozdrenko, Dmytro; Prylutska, Svitlana; Bogutska, Kateryna; Nurishchenko, Natalia Y.; Abramchuk, Olga; Motuziuk, Olexandr; Prylutskyy, Yuriy; Scharff, Peter; Ritter, Uwe
Effect of C60 fullerene on recovery of muscle soleus in rats after atrophy induced by achillotenotomy. - In: Life, ISSN 2075-1729, Bd. 12 (2022), 3, 332, S. 1-13

Biomechanical and biochemical changes in the muscle soleus of rats during imitation of hind limbs unuse were studied in the model of the Achilles tendon rupture (Achillotenotomy). Oral administration of water-soluble C60 fullerene at a dose of 1 mg/kg was used as a therapeutic agent throughout the experiment. Changes in the force of contraction and the integrated power of the muscle, the time to reach the maximum force response, the mechanics of fatigue processes development, in particular, the transition from dentate to smooth tetanus, as well as the levels of pro- and antioxidant balance in the blood of rats on days 15, 30 and 45 after injury were described. The obtained results indicate a promising prospect for C60 fullerene use as a powerful antioxidant for reducing and correcting pathological conditions of the muscular system arising from skeletal muscle atrophy.



https://doi.org/10.3390/life12030332
Mytiliniou, Maria; Wondergem, Joeri A. J.; Schmidt, Thomas; Heinrich, Doris
Impact of neurite alignment on organelle motion. - In: Interface, ISSN 1742-5662, Bd. 19 (2022), 187, 20210617, S. 1-13

Intracellular transport is pivotal for cell growth and survival. Malfunctions in this process have been associated with devastating neurodegenerative diseases, highlighting the need for a deeper understanding of the mechanisms involved. Here, we use an experimental methodology that leads neurites of differentiated PC12 cells into either one of two configurations: a one-dimensional configuration, where the neurites align along lines, or a two-dimensional configuration, where the neurites adopt a random orientation and shape on a flat substrate. We subsequently monitored the motion of functional organelles, the lysosomes, inside the neurites. Implementing a time-resolved analysis of the mean-squared displacement, we quantitatively characterized distinct motion modes of the lysosomes. Our results indicate that neurite alignment gives rise to faster diffusive and super-diffusive lysosomal motion than the situation in which the neurites are randomly oriented. After inducing lysosome swelling through an osmotic challenge by sucrose, we confirmed the predicted slowdown in diffusive mobility. Surprisingly, we found that the swelling-induced mobility change affected each of the (sub-/super-)diffusive motion modes differently and depended on the alignment configuration of the neurites. Our findings imply that intracellular transport is significantly and robustly dependent on cell morphology, which might in part be controlled by the extracellular matrix.



https://doi.org/10.1098/rsif.2021.0617
Köhler, Michael;
Vaccination, immunity and breakthrough: quantitative effects in individual immune responses illustrated by a simple kinetic model. - In: Applied Sciences, ISSN 2076-3417, Bd. 12 (2022), 1, 31, S. 1-15

The personal risks of infection, as well as the conditions for achieving herd immunity, are strongly dependent on an individual’s response to the infective agents on the one hand, and the individual’s reactions to vaccination on the other hand. The main goal of this work is to illustrate the importance of quantitative individual effects for disease risk in a simple way. The applied model was able to illustrate the quantitative effects, in the cases of different individual reactions, after exposition to viruses or bacteria and vaccines. The model was based on simple kinetic equations for stimulation of antibody production using different concentrations of the infective agent, vaccine and antibodies. It gave a qualitative explanation for the individual differences in breakthrough risks and different requirements concerning a second, third or further vaccinations, reconsidering different efficiencies of the stimulation of an immune reaction.



https://doi.org/10.3390/app12010031