Scientific publications

2018
Exploring the potential of electroless and electroplated noble metal–semiconductor hybrids within bio- and environmental sensing

Semenova, Daria | Gernaey, K. V. | Silina, Yuliya E.

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Over the last two decades, the rapid development and widespread application of nanomaterials has significantly influenced research in various fields, including analytical chemistry and biosensing technologies. In particular, the simple functionalization and tuning of noble metal nanoparticle (NP) surface chemistry resulted in the development of a series of novel biosensing platforms with quick read-out and enhanced capabilities towards specific analyte detection. Moreover, noble metal NPs possess a number of unique properties, viz. high surface-to-volume ratio and excellent spectral, optical, thermal, electrical and catalytic characteristics. This manuscript provides an elaborate review on galvanic noble metal NPs deposited onto semiconductor surfaces, from the preparation stage towards their application in biosensors and gas sensing. Two types of deposition approaches, viz. galvanic displacement/electroless and conventional electroplating, are introduced and compared. Furthermore, the analytical merit of hybrid nanomaterials towards the improvement of sensing abilities is highlighted. Finally, some limitations and challenges related to progress in the development and application of analytical devices based on electroless and electroplated noble metal NPs-semiconductor hybrids (NMNPsHs) in biochemical and environmental sensing are discussed.

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Analyst,
2018, 143 (23), 5646-5669.

Exploring the Potential of Electroplated Chips towards Biomedical Sensing and Diagnostics

Semenova, Daria | Silina, Yuliya E.

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In the past decade the significant progress in the cellular stress response was witnessed. Nevertheless, the development of the minimally-invasive and accurate sensing tools for the identification of the increasing number of potentially relevant species in clinical diagnostics, using smaller sample volumes is a major challenge. Herein, the potential of the electroplated nanomaterials towards biomedical sensing and diagnostics is summarized. The key factors affecting the surface functionality, dimensionality, S/N ratio and analytical response of the prepared chips are highlighted. Furthermore, the application of electroplated chips as a fast “read out” platform for profiling of clinical samples was demonstrated.

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Proceedings,
2018, 2 (13), 817.

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Human Co- and Triple-Culture Model of the Alveolar-Capillary Barrier on a Basement Membrane Mimic

Dohle, Eva | Singh, Smriti | Nishigushi, Akihiro | Fischer, Thorsten | Wessling, Matthias | Möller, Martin | Sader, Robert | Kasper, Jennifer Y. | Ghanaati, Shahram | Kirkpatrick, C. James

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The development of an in vitro model resembling the alveolar-capillary barrier might be a highly beneficial tool to study lung physiology as well as the immune response of the lung to infection or after exposure to nanoparticles. This study is based on an in vitro alveolar barrier developed on a basement membrane mimic, composed of ultrathin nanofiber meshes generated via electrospinning using bioresorbable poly(ɛ-caprolactone). As cellular components, NCI H441, resembling the alveolar epithelial cells, and ISO-HAS-1, an endothelial cell line, were used to perform bipolar coculture experiments for a total cultivation period of 14 days. In addition to immunohistochemical and immunofluorescent studies, transepithelial electrical resistance (TER) and transport capabilities of the in vitro model system were investigated. Alveolar barrier function could be clearly determined for the postulated bipolar coculture system on the basement membrane mimic, since TER increased during the course of bipolar cultivation. Furthermore, to gain first insights into possible lung inflammatory reactions in vitro, this coculture model was further expanded by a human leukemia monocyte cell line (THP-1). This triple-culture system was able to maintain adequately the barrier properties of the bipolar coculture, thus making this in vitro model consisting of epithelial, endothelial, and immune cells on a basement membrane mimic a promising basis for further studies in tissue engineering.

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Tissue Engineering Part C: Methods,
2018, 24 (9), 495-503.

Photoactivatable Adhesive Ligands for Light-Guided Neuronal Growth

Farrukh, Aleeza | Fan, Wenqiang | Zhao, Shifang | Salierno, Marcelo | Paez, Julieta I. | Del Campo, Aranzazu

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Abstract Neuro-regeneration after trauma requires growth and reconnection of neurons to reestablish information flow in particular directions across the damaged tissue. To support this process, biomaterials for nerve tissue regeneration need to provide spatial information to adhesion receptors on the cell membrane and to provide directionality to growing neurites. Here, photoactivatable adhesive peptides based on the CASIKVAVSADR laminin peptidomimetic are presented and applied to spatiotemporal control of neuronal growth to biomaterials in vitro. The introduction of a photoremovable group [6-nitroveratryl (NVOC), 3-(4,5-dimethoxy-2-nitrophenyl)butan-2-yl (DMNPB), or 2,2′-((3′-(1-hydroxypropan-2-yl)-4′-nitro-[1,1′-biphenyl]-4-yl)azanediyl)bis(ethan-1-ol) (HANBP)] at the amino terminal group of the K residue temporally inhibited the activity of the peptide. The bioactivity was regained through controlled light exposure. When used in neuronal culture substrates, the peptides allowed light-based control of the attachment and differentiation of neuronal cells. Site-selective irradiation activated adhesion and differentiation cues and guided seeded neurons to grow in predefined patterns. This is the first demonstration of ligand-based light-controlled interaction between neuronal cells and biomaterials.

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ChemBioChem,
2018, 19 (12), 1271-1279.

Microenvironments Designed to Support Growth and Function of Neuronal Cells

Farrukh, Aleeza | Zhao, Shifang | del Campo, Aránzazu

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Strategies for neural tissue repair heavily depend on our ability to temporally reconstruct the natural cellular microenvironment of neural cells. Biomaterials play a fundamental role in this context, as they provide the mechanical support for cells to attach and migrate to the injury site, as well as fundamental signals for differentiation. This review describes how different cellular processes (attachment, proliferation, and (directional) migration and differentiation) have been supported by different material parameters, in vitro and in vivo. Although incipient guidelines for biomaterial design become visible, literature in the field remains rather phenomenological. As in other fields of tissue regeneration, progress will depend on more systematic studies on cell-materials response, better understanding on how cells behave and understand signals in their natural milieu from neurobiology studies, and the translation of this knowledge into engineered microenvironments for clinical use.

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Frontiers in Materials,
2018, 5 (62).

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In Situ, Light-Guided Axon Growth on Biomaterials via Photoactivatable Laminin Peptidomimetic IK(HANBP)VAV

Farrukh, Aleeza | Zhao, Shifang | Paez, Julieta I. | Kavyanifar, Atria | Salierno, Marcelo | Cavalié, Adolfo | del Campo, Aránzazu

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The ability to guide the growth of neurites is relevant for reconstructing neural networks and for nerve tissue regeneration. Here, a biofunctional hydrogel that allows light-based directional control of axon growth in situ is presented. The gel is covalently modified with a photoactivatable derivative of the short laminin peptidomimetic IKVAV. This adhesive peptide contains the photoremovable group 2-(4′-amino-4-nitro-[1,1′-biphenyl]-3-yl)propan-1-ol (HANBP) on the Lys rest that inhibits its activity. The modified peptide is highly soluble in water and can be simply conjugated to −COOH containing hydrogels via its terminal −NH2 group. Light exposure allows presentation of the IKVAV adhesive motif on a soft hydrogel at desired concentration and at defined position and time point. The photoactivated gel supports neurite outgrowth in embryonic neural progenitor cells culture and allows site-selective guidance of neurites extension. In situ exposure of cell cultures using a scanning laser allows outgrowth of neurites in desired pathways.

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ACS Applied Materials & Interfaces,
2018, 10 (48), 41129-41137.

A Photoactivatable α5β1-Specific Integrin Ligand

Nair, Roshna V. | Farrukh, Aleeza | del Campo, Aránzazu

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Abstract The integrin α5β1 is overexpressed in colon, breast, ovarian, lung and brain tumours, and has been identified as key component in mechanosensing. In order to study how dynamic changes in α5β1 engagement affect cellular behaviour, photoactivatable derivatives of α5β1-specific ligands are presented in this article. A photoremovable protecting group (PRPG) was introduced into the ligand structure at a relevant position for integrin recognition. The presence of the chromophore temporarily inhibited ligand bioactivity. Light exposure at a cell-compatible dose efficiently cleaved the protecting group and restored functionality. The photoactive ligand had an azide end-functional group for covalent immobilisation onto biomaterials by click chemistry. Selective cell responses (attachment, spreading, migration) to the activated ligand on the surface are achieved by controlled exposure to light, at similar levels to the native ligand. Spatial and temporal control of the cellular response is demonstrated, including the possibility of in situ activation. Photoactivatable integrin-selective ligands in model microenvironments will allow the study of cellular behaviour in response to changes in the activation of individual integrins as consequence of dynamic variations in matrix composition.

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ChemBioChem,
2018, 19 (12), 1280-1287.

A bio-based route to the carbon-5 chemical glutaric acid and to bionylon-6,5 using metabolically engineered Corynebacterium glutamicum

Rohles, Christina Maria | Gläser, Lars | Kohlstedt, Michael | Gießelmann, Gideon | Pearson, Samuel | del Campo, Aránzazu | Becker, Judith | Wittmann, Christoph

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In the present work, we established the bio-based production of glutarate, a carbon-5 dicarboxylic acid with recognized value for commercial plastics and other applications, using metabolically engineered Corynebacterium glutamicum. The mutant C. glutamicum AVA-2 served as a starting point for strain development, because it secreted small amounts of glutarate as a consequence of its engineered 5-aminovalerate pathway. Starting from AVA-2, we overexpressed 5-aminovalerate transaminase (gabT) and glutarate semialdehyde dehydrogenase (gabD) under the control of the constitutive tuf promoter to convert 5-aminovalerate further to glutarate. The created strain GTA-1 formed glutarate as a major product, but still secreted 5-aminovalerate as well. This bottleneck was tackled at the level of 5-aminovalerate re-import. The advanced strain GTA-4 overexpressed the newly discovered 5-aminovalerate importer NCgl0464 and formed glutarate from glucose in a yield of 0.27 mol mol−1. In a fed-batch process, GTA-4 produced more than 90 g L−1 glutarate from glucose and molasses based sugars in a yield of up to 0.70 mol mol−1 and a maximum productivity of 1.8 g L−1 h−1, while 5-aminovalerate was no longer secreted. The bio-based glutaric acid was purified to >99.9% purity. Interfacial polymerization and melt polymerization with hexamethylenediamine yielded bionylon-6,5, a polyamide with a unique structu

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Green Chemistry,
2018, 20 (20), 4662-4674.

OPEN ACCESS
Toward Light-Regulated Living Biomaterials

Sankaran, Shrikrishnan | Zhao, Shifang | Muth, Christina | Paez, Julieta | del Campo, Aránzazu

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Abstract Living materials are an emergent material class, infused with the productive, adaptive, and regenerative properties of living organisms. Property regulation in living materials requires encoding responsive units in the living components to allow external manipulation of their function. Here, an optoregulated Escherichia coli (E. coli)-based living biomaterial that can be externally addressed using light to interact with mammalian cells is demonstrated. This is achieved by using a photoactivatable inducer of gene expression and bacterial surface display technology to present an integrin-specific miniprotein on the outer membrane of an endotoxin-free E. coli strain. Hydrogel surfaces functionalized with the bacteria can expose cell adhesive molecules upon in situ light-activation, and trigger cell adhesion. Surface immobilized bacteria are able to deliver a fluorescent protein to the mammalian cells with which they are interacting, indicating the potential of such a bacterial material to deliver molecules to cells in a targeted manner.

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Advanced Science,
2018, 5 (8), 1800383.

OPEN ACCESS
Thermal decomposition of the amino acids glycine, cysteine, aspartic acid, asparagine, glutamic acid, glutamine, arginine and histidine

Weiss, Ingrid M. | Muth, Christina | Drumm, Robert | Kirchner, Helmut O. K.

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The pathways of thermal instability of amino acids have been unknown. New mass spectrometric data allow unequivocal quantitative identification of the decomposition products.

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BMC Biophysics,
2018, 11 (1), 2.

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