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- ItemGlycerylphytate as an ionic crosslinker for 3D printing of multi-layered scaffolds with improved shape fidelity and biological features(London : Royal Society of Chemistry, 2020) Mora-Boza, A.; Włodarczyk-Biegun, M.K.; Del Campo, A.; Vázquez-Lasa, B.; Román, J.S.The fabrication of intricate and long-term stable 3D polymeric scaffolds by a 3D printing technique is still a challenge. In the biomedical field, hydrogel materials are very frequently used because of their excellent biocompatibility and biodegradability, however the improvement of their processability and mechanical properties is still required. This paper reports the fabrication of dual crosslinked 3D scaffolds using a low concentrated (<10 wt%) ink of gelatin methacryloyl (GelMA)/chitosan and a novel crosslinking agent, glycerylphytate (G1Phy) to overcome the current limitations in the 3D printing field using hydrogels. The applied methodology consisted of a first ultraviolet light (UV) photopolymerization followed by a post-printing ionic crosslinking treatment with G1Phy. This crosslinker provides a robust framework and avoids the necessity of neutralization with strong bases. The blend ink showed shear-thinning behavior and excellent printability in the form of a straight and homogeneous filament. UV curing was undertaken simultaneously to 3D deposition, which enhanced precision and shape fidelity (resolution ≈150 μm), and prevented the collapse of the subsequent printed layers (up to 28 layers). In the second step, the novel G1Phy ionic crosslinker agent provided swelling and long term stability properties to the 3D scaffolds. The multi-layered printed scaffolds were mechanically stable under physiological conditions for at least one month. Preliminary in vitro assays using L929 fibroblasts showed very promising results in terms of adhesion, spreading, and proliferation in comparison to other phosphate-based traditional crosslinkers (i.e. TPP). We envision that the proposed combination of the blend ink and 3D printing approach can have widespread applications in the regeneration of soft tissues.
- ItemTime-resolved luminescence detection of peroxynitrite using a reactivity-based lanthanide probe(Cambridge : RSC, 2020) Breen, Colum; Pal, Robert; Elsegood, Mark R.J.; Teat, Simon J.; Iza, Felipe; Wende, Kristian; Buckley, Benjamin R.; Butler, StephenPeroxynitrite (ONOO-) is a powerful and short-lived oxidant formed in vivo, which can react with most biomolecules directly. To fully understand the roles of ONOO- in cell biology, improved methods for the selective detection and real-time analysis of ONOO- are needed. We present a water-soluble, luminescent europium(iii) probe for the rapid and sensitive detection of peroxynitrite in human serum, living cells and biological matrices. We have utilised the long luminescence lifetime of the probe to measure ONOO- in a time-resolved manner, effectively avoiding the influence of autofluorescence in biological samples. To demonstrate the utility of the Eu(iii) probe, we monitored the production of ONOO- in different cell lines, following treatment with a cold atmospheric plasma device commonly used in the clinic for skin wound treatment. This journal is © The Royal Society of Chemistry.
- ItemA non-cytotoxic resin for micro-stereolithography for cell cultures of HUVECs(Basel : MDPI, 2020) Männel, Max J.; Fischer, Carolin; Thiele, JulianThree-dimensional (3D) printing of microfluidic devices continuously replaces conventional fabrication methods. A versatile tool for achieving microscopic feature sizes and short process times is micro-stereolithography (µSL). However, common resins for µSL lack biocompatibility and are cytotoxic. This work focuses on developing new photo-curable resins as a basis for µSL fabrication of polymer materials and surfaces for cell culture. Different acrylate-and methacrylate-based compositions are screened for material characteristics including wettability, surface roughness, and swelling behavior. For further understanding, the impact of photo-absorber and photo-initiator on the cytotoxicity of 3D-printed substrates is studied. Cell culture experiments with human umbilical vein endothelial cells (HUVECs) in standard polystyrene vessels are compared to 3D-printed parts made from our library of homemade resins. Among these, after optimizing material composition and post-processing, we identify selected mixtures of poly(ethylene glycol) diacrylate (PEGDA) and poly(ethylene glycol) methyl ethyl methacrylate (PEGMEMA) as most suitable to allow for fabricating cell culture platforms that retain both the viability and proliferation of HUVECs. Next, our PEGDA/PEGMEMA resins will be further optimized regarding minimal feature size and cell adhesion to fabricate microscopic (microfluidic) cell culture platforms, e.g., for studying vascularization of HUVECs in vitro. © 2020 by the authors.
- ItemBlood platelet enrichment in mass-producible surface acoustic wave (SAW) driven microfluidic chips(Cambridge : RSC, 2019) Richard, Cynthia; Fakhfouri, Armaghan; Colditz, Melanie; Striggow, Friedrich; Kronstein-Wiedemann, Romy; Tonn, Torsten; Medina-Sánchez, Mariana; Schmidt, Oliver G.; Gemming, Thomas; Winkler, AndreasThe ability to separate specific biological components from cell suspensions is indispensable for liquid biopsies, and for personalized diagnostics and therapy. This paper describes an advanced surface acoustic wave (SAW) based device designed for the enrichment of platelets (PLTs) from a dispersion of PLTs and red blood cells (RBCs) at whole blood concentrations, opening new possibilities for diverse applications involving cell manipulation with high throughput. The device is made of patterned SU-8 photoresist that is lithographically defined on the wafer scale with a new proposed methodology. The blood cells are initially focused and subsequently separated by an acoustic radiation force (ARF) applied through standing SAWs (SSAWs). By means of flow cytometric analysis, the PLT concentration factor was found to be 7.7, and it was proven that the PLTs maintain their initial state. A substantially higher cell throughput and considerably lower applied powers than comparable devices from literature were achieved. In addition, fully coupled 3D numerical simulations based on SAW wave field measurements were carried out to anticipate the coupling of the wave field into the fluid, and to obtain the resulting pressure field. A comparison to the acoustically simpler case of PDMS channel walls is given. The simulated results show an ideal match to the experimental observations and offer the first insights into the acoustic behavior of SU-8 as channel wall material. The proposed device is compatible with current (Lab-on-a-Chip) microfabrication techniques allowing for mass-scale, reproducible chip manufacturing which is crucial to push the technology from lab-based to real-world applications. © The Royal Society of Chemistry.
- ItemThe LEGATO cross-disciplinary integrated ecosystem service research framework: an example of integrating research results from the analysis of global change impacts and the social, cultural and economic system dynamics of irrigated rice production(Heidelberg : Springer Verlag, 2017) Spangenberg, J.H.; Beaurepaire, A.L.; Bergmeier, E.; Burkhard, B.; van Chien, H.; Cuong, L.Q.; Görg, C.; Grescho, V.; Hai, L.H.; Heong, K.L.; Horgan, F.G.; Hotes, S.; Klotzbücher, A.; Klotzbücher, T.; Kühn, I.; Langerwisch, F.; Marion, G.; Moritz, R.F.A.; Nguyen, Q.A.; Ott, J.; Sann, C.; Sattler, C.; Schädler, M.; Schmidt, A.; Tekken, V.; Thanh, T.D.; Thonicke, K.; Türke, M.; VáclavÃk, T.; Vetterlein, D.; Westphal, C.; Wiemers, M.; Settele, J.In a cross-disciplinary project (LEGATO) combining inter- and transdisciplinary methods, we quantify the dependency of rice-dominated socio-ecological systems on ecosystem functions (ESF) and the ecosystem services (ESS) the integrated system provides. In the collaboration of a large team including geo- and bioscientists, economists, political and cultural scientists, the mutual influences of the biological, climate and soil conditions of the agricultural area and its surrounding natural landscape have been analysed. One focus was on sociocultural and economic backgrounds, another on local as well as regional land use intensity and biodiversity, and the potential impacts of future climate and land use change. LEGATO analysed characteristic elements of three service strands defined by the Millennium Ecosystem Assessment (MA): (a) provisioning services: nutrient cycling and crop production; (b) regulating services: biocontrol and pollination; and (c) cultural services: cultural identity and aesthetics. However, in line with much of the current ESS literature, what the MA called supporting services is treated as ESF within LEGATO. As a core output, LEGATO developed generally applicable principles of ecological engineering (EE), suitable for application in the context of future climate and land use change. EE is an emerging discipline, concerned with the design, monitoring and construction of ecosystems and aims at developing strategies to optimise ecosystem services through exploiting natural regulation mechanisms instead of suppressing them. Along these lines LEGATO also aims to create the knowledge base for decision-making for sustainable land management and livelihoods, including the provision of the corresponding governance and management strategies, technologies and system solutions.
- ItemRaman-spectroscopy based cell identification on a microhole array chip(Basel : MDPI AG, 2014) Neugebauer, U.; Kurz, C.; Bocklitz, T.; Berger, T.; Velten, T.; Clement, J.H.; Krafft, C.; Popp, J.Circulating tumor cells (CTCs) from blood of cancer patients are valuable prognostic markers and enable monitoring responses to therapy. The extremely low number of CTCs makes their isolation and characterization a major technological challenge. For label-free cell identification a novel combination of Raman spectroscopy with a microhole array platform is described that is expected to support high-throughput and multiplex analyses. Raman spectra were registered from regularly arranged cells on the chip with low background noise from the silicon nitride chip membrane. A classification model was trained to distinguish leukocytes from myeloblasts (OCI-AML3) and breast cancer cells (MCF-7 and BT-20). The model was validated by Raman spectra of a mixed cell population. The high spectral quality, low destructivity and high classification accuracy suggests that this approach is promising for Raman activated cell sorting.
- ItemRolled-up functionalized nanomembranes as three-dimensional cavities for single cell studies(Washington, DC : American Chemical Society, 2014) Xi, W.; Schmidt, C.K.; Sanchez, S.; Gracias, D.H.; Carazo-Salas, R.E.; Jackson, S.P.; Schmidt, O.G.We use micropatterning and strain engineering to encapsulate single living mammalian cells into transparent tubular architectures consisting of three-dimensional (3D) rolled-up nanomembranes. By using optical microscopy, we demonstrate that these structures are suitable for the scrutiny of cellular dynamics within confined 3D-microenvironments. We show that spatial confinement of mitotic mammalian cells inside tubular architectures can perturb metaphase plate formation, delay mitotic progression, and cause chromosomal instability in both a transformed and nontransformed human cell line. These findings could provide important clues into how spatial constraints dictate cellular behavior and function.
- ItemA novel patch micro electrode array for sensing ionic membrane currents(Amsterdam [u.a.] : Elsevier, 2011) Aryasomayajula, A.; Perike, S.; Hensel, R.; Posseckardt, J.; Gerlach, G.; Funk, R.H.W.Ionic membrane currents play an important role during regeneration of nerve cells, embryonic development and wound healing processes. Measuring the intracellular ion currents across the cell membrane is important in understanding the cellular functions related to the ion activities. A novel patch micro electrode array (p-MEA) for measuring the ionic membrane currents without poisoning the cells due to emitting metal ions is described in this paper. Results on biocompatibility of the device are presented. We discuss the fabrication and working principle of p-MEA.