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Glycerylphytate as an ionic crosslinker for 3D printing of multi-layered scaffolds with improved shape fidelity and biological features

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.

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Rolled-up functionalized nanomembranes as three-dimensional cavities for single cell studies

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.

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The 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

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., Václavík, 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.

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A novel patch micro electrode array for sensing ionic membrane currents

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.

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Raman-spectroscopy based cell identification on a microhole array chip

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.