BIOGEL

A large library of building blocks was established, ranging from the molecular level, such as bicyclic peptides, engineered elastin-like recombinamers (ELRs), and modified polymers, to microscale elements, such as microgels. A combination of these biological and synthetic building blocks were applied to construct hybrid macroscopic biointegrative coatings, tissue engineering constructs, and diagnostic devices. The bottom-up synthetic approach to create these novel materials enables easy adjustment for a wide range of applications. Within the project, we discovered new types of highly selective bicyclic peptides for cell adhesion, which were incorporated in 3 different types of three-dimensional (3D) hydrogels, leading to improved cell spreading and growth. Regenerative materials are important to heal damaged or injured tissues in the body. One example is articular cartilage injury, which is a serious clinical problem as it can lead to disabilities and high health care costs when untreated. Within Biogel, different injectable gels were designed to fill irregular cavities and support cell growth and regeneration. However, many hydrogels for 3D cell culture do not efficiently mimic the biological environment of a cell, including the open porous architecture and the mechanical properties. Here, we developed dynamic hydrogels, which can adjust to the needs of the cells and growing tissues, and were employed to study the mechanobiology involved in the interaction between cells and their substrates. In addition to larger hydrogels, small cell-loaden micron-scale microgels were produced, which can be injected as cell transplant. The microgels protect the cells and reduce their migration away from the injury site. In addition to 3D regenerative materials, medical polymer coatings were tested to improve the efficiency of glucose sensors. Moreover, new types of hydrogel-based biosensors were developed that comprise metallic nanostructures and responsive hydrogel architectures to target analytes present in complex biological fluids.

(i) Different coating systems were developed that do not alter the function of the sensor They retain glucose sensitivity, while allowing the introduction of non-fouling behavior (doi: 10.1021/acs.biomac.7b00516 10.1073/pnas.1720055115 10.1021/acs.biomac.8b01405). Stable cholesteryl modified ELR-based nano-carriers were produced with superior cell-membrane interaction and cell-type specificity for potential cell coatings or drug delivery systems.

(ii) More efficient cell adhesive bicyclic peptides were synthesized (doi: 10.1021/acscombsci.8b00144) with a newly developed technique to measure binding efficiency (doi: 10.1021/acs.analchem.7b00554). For the purpose of cartilage repair, two injectable hydrogels were developed: One is a hybrid Silk Elastin-Like co-Recombinamer hydrogel (doi: 10.1021/acs.biomac.8b01211) which can be post-modified with the bicyclic peptides yielding in enhanced cell adhesion (doi: 10.1088/1748-605X/aafd83). The second gel is an injectable hyaluronic acid-based hydrogel with tunable biochemical and mechanical properties (doi: 10.1016/j.eurpolymj.2019.02.024). In addition to these biological gels, synthetic gels were developed to better mimic the non-linear mechanics of the natural ECM. Fibrous, macroporous polyisocyanides (PIC) hydrogels were synthesized with stress-stiffening behavior, regulating cell behavior (doi: 10.1021/acs.biomac.8b01445). A combination of PIC with the bicyclic peptides accelerates stem cell spreading (doi: 10.1038/s41467-019-08569-4). The mechanical properties and gelation temperature can be tailored by copolymerization (doi: 10.1016/j.cclet.2017.11.002). Another method to obtain macropores in an injectable material is by crosslinking micron-scale microgels together after injection. Novel microfluidic techniques were established to produce hollow and rod-shaped microgels that enable drug and cell encapsulation directly on chip (doi: 10.1039/c7bm00322f). In order to mimic the tissue architecture, bioactive domains are patterned into 3D poly(ethylene glycol) hydrogels using light-sensitive chemistry. To test the efficiency of the novel Biogel hydrogels ex vivo, a new bioreactor platform was created. Nanocellulose-based hydrogels were successfully employed to culture and differentiate human liver organoids. In addition, an isolated perfused porcine liver platform was realized.

(iii) Poly (N-isopropylacrylamide) (pNIPAM)-based thermosensitive hydrogels, embedded with gold nanoparticles, were prepared as hydrogel films that can locally actuate upon irradiation to study mechanobiology. Nanopatterned pNIPAM microgel surfaces leads to controlled bending of microgels upon actuation. The same pNIPAM/gold material was employed to design new diagnostic devices with improved sensitivity (doi: 10.1039/C7NR08905H). In parallel, a portable optical biosensor platform was established for label-free detection of proteins. Additionally, ELRs with both a lower and upper critical solution temperature were produced with potential use as responsive temperature sensors. A last sensing approach targeted the degradability of specific peptide patterns incorporated into the ELR backbone to detect different proteases.

To improve biointegration of medical devices and sensors inside the body and increase their life-time, Biogel developed new hydrogel coatings. As tissue engineering application, cartilage repair was a main focus to reduce disability and high costs of treatment. The newly developed injectable cell-laden hydrogels can be administrated via a minimally invasive procedure and fill irregularly-shaped cartilage defects, combined with bioactive molecules and anti-inflammatory drugs. Besides cartilage repair, many other damaged or diseased tissues could greatly benefit from efficient regeneration therapies. The challenge is to design materials that provide the right signals to form functional tissue. The artificial synthetic polymeric hydrogels developed in Biogel better mimic the extracellular matrix, have tunable properties to direct cellular responses, and can be mixed with other gels to to obtain properties that cannot be achieved using single component gels. A novel microfluidic technique was developed to also encapsulate drugs and cells inside microgels to combine cell transplantations with material-based therapies to increase their efficiency. As testing platform, a new bioreactor was developed that allows for long-term flow cultures of cells within hydrogel constructs with online analysis methods. With this technology, tissues can be constructed that mimic the in vivo situation to gain relevant insights into the mechanisms of the cells or pathologies thereof. Besides the bioreactor, an ex vivo perfusion system was established to keep pig livers alive and develop a damage model of a fatty liver to explore the mechanisms involved in this illness and test possible cures. For diagnostic purposes, tailored hydrogel materials are deployed at the interface between the biological sample and technical transducer to open new ways for a sensitive analysis of trace amounts of biomarkers in new settings that are closer to the patient and do not require centralized laboratories. These advances allow for portable devices and the detection of lower target concentrations, which will in the future pave the way to detect earlier stages of diseases.

Summer School
5 day Summer School to focus TP8: general skills (iv-x)

Workshop 1
2 day Workshop 1: TP8: general skills (i-iii)

Workshop 2
3 day Workshop 2: TP1: Synthesis

Workshop 3
3 day Workshop 3: TP2: Mechanical characterization

Workshop 4
3 day Workshop 4: TP3: Biomimetic hydrogels

Ambassador Day
All ESRs will act as Marie Skłodowska-Curie Ambassadors and visit schools, universities, community organizations

International conference
3 day international conference organized by BIOGEL

Workshop 5
3 day Workshop 5: TP4: Coatings and sensors

Workshop 6
4 day Workshop 6: TP5-6: in vitro, ex vivo, and in vivo models

Workshop 7
3 day Workshop 7: TP7: Imaging

Open ITN day
All participants will open their door to inform the public one day

Bicyclic RGD Peptides with Exquisite Selectivity for the Integrin α v β 3 Receptor Using a “Random Design” Approach
Author(s): Dominik Bernhagen, Vanessa Jungbluth, Nestor Gisbert Quilis, Jakub Dostalek, Paul B. White, Kees Jalink, Peter Timmerman
Published in: ACS Combinatorial Science, 2019, ISSN 2156-8952
DOI: 10.1021/acscombsci.8b00144

Cytoskeletal stiffening in synthetic hydrogels
Author(s): Paula de Almeida, Maarten Jaspers, Sarah Vaessen, Oya Tagit, Giuseppe Portale, Alan E. Rowan, Paul H. J. Kouwer
Published in: Nature Communications, Issue 10/1, 2019, Page(s) 609, ISSN 2041-1723
DOI: 10.1038/s41467-019-08569-4

Synthetic Extracellular Matrices with Nonlinear Elasticity Regulate Cellular Organization
Author(s): Kaizheng Liu, Silvia M. Mihaila, Alan Rowan, Egbert Oosterwijk, Paul H. J. Kouwer
Published in: Biomacromolecules, Issue 20/2, 2019, Page(s) 826-834, ISSN 1525-7797
DOI: 10.1021/acs.biomac.8b01445

Controlling the gelation temperature of biomimetic polyisocyanides
Author(s): Paul H.J. Kouwer, Paula de Almeida, Onno ven den Boomen, Zaskia H. Eksteen-Akeroyd, Roel Hammink, Maarten Jaspers, Stijn Kragt, Mathijs F.J. Mabesoone, Roeland J.M. Nolte, Alan E. Rowan, Martin G.T.A. Rutten, Vincent A.A. Le Sage, Daniël C. Schoenmakers, Chengfen Xing, Jialiang Xu
Published in: Chinese Chemical Letters, Issue 29/2, 2018, Page(s) 281-284, ISSN 1001-8417
DOI: 10.1016/j.cclet.2017.11.002

Polymer Brush-Functionalized Chitosan Hydrogels as Antifouling Implant Coatings
Author(s): Irene Buzzacchera, Mariia Vorobii, Nina Yu. Kostina, Andres de los Santos Pereira, Tomáš Riedel, Michael Bruns, Wojciech Ogieglo, Martin Möller, Christopher J. Wilson, Cesar Rodriguez-Emmenegger
Published in: Biomacromolecules, Issue 18/6, 2017, Page(s) 1983-1992, ISSN 1525-7797
DOI: 10.1021/acs.biomac.7b00516

Screening Libraries of Amphiphilic Janus Dendrimers Based on Natural Phenolic Acids to Discover Monodisperse Unilamellar Dendrimersomes
Author(s): Irene Buzzacchera, Qi Xiao, Hong Han, Khosrow Rahimi, Shangda Li, Nina Yu. Kostina, B. Jelle Toebes, Samantha E. Wilner, Martin Möller, Cesar Rodriguez-Emmenegger, Tobias Baumgart, Daniela A. Wilson, Christopher J. Wilson, Michael L. Klein, Virgil Percec
Published in: Biomacromolecules, Issue 20/2, 2019, Page(s) 712-727, ISSN 1525-7797
DOI: 10.1021/acs.biomac.8b01405

Microfluidic fabrication of polyethylene glycol microgel capsules with tailored properties for the delivery of biomolecules
Author(s): Luis P. B. Guerzoni, Jan Bohl, Alexander Jans, Jonas C. Rose, Jens Koehler, Alexander J. C. Kuehne, Laura De Laporte
Published in: Biomaterials Science, Issue 5/8, 2017, Page(s) 1549-1557, ISSN 2047-4830
DOI: 10.1039/c7bm00322f

Clinical Evidence for Use of a Noninvasive Biosensor for Tear Glucose as an Alternative to Painful Finger-Prick for Diabetes Management Utilizing a Biopolymer Coating
Author(s): Alicja E. Kownacka, Dovile Vegelyte, Maurits Joosse, Nicoleta Anton, B. Jelle Toebes, Jan Lauko, Irene Buzzacchera, Katarzyna Lipinska, Daniela A. Wilson, Nel Geelhoed-Duijvestijn, Christopher J. Wilson
Published in: Biomacromolecules, Issue 19/11, 2018, Page(s) 4504-4511, ISSN 1525-7797
DOI: 10.1021/acs.biomac.8b01429

On the synthesis and characterization of biofunctional hyaluronic acid based injectable hydrogels for the repair of cartilage lesions
Author(s): Evgenia Tsanaktsidou, Olga Kammona, Costas Kiparissides
Published in: European Polymer Journal, Issue 114, 2019, Page(s) 47-56, ISSN 0014-3057
DOI: 10.1016/j.eurpolymj.2019.02.024

Tunable laser interference lithography preparation of plasmonic nanoparticle arrays tailored for SERS
Author(s): Nestor Gisbert Quilis, Médéric Lequeux, Priyamvada Venugopalan, Imran Khan, Wolfgang Knoll, Souhir Boujday, Marc Lamy de la Chapelle, Jakub Dostalek
Published in: Nanoscale, Issue 10/21, 2018, Page(s) 10268-10276, ISSN 2040-3364
DOI: 10.1039/c7nr08905h

Encoding biological recognition in a bicomponent cell-membrane mimic
Author(s): Cesar Rodriguez-Emmenegger, Qi Xiao, Nina Yu. Kostina, Samuel E. Sherman, Khosrow Rahimi, Benjamin E. Partridge, Shangda Li, Dipankar Sahoo, Aracelee M. Reveron Perez, Irene Buzzacchera, Hong Han, Meir Kerzner, Ishita Malhotra, Martin Möller, Christopher J. Wilson, Matthew C. Good, Mark Goulian, Tobias Baumgart, Michael L. Klein, Virgil Percec
Published in: Proceedings of the National Academy of Sciences, 2017, Page(s) 201821924, ISSN 0027-8424
DOI: 10.1073/pnas.1821924116

High-Affinity RGD-Knottin Peptide as a New Tool for Rapid Evaluation of the Binding Strength of Unlabeled RGD-Peptides to α v β 3 , α v β 5 , and α 5 β 1 Integrin Receptors
Author(s): Dominik Bernhagen, Laura De Laporte, Peter Timmerman
Published in: Analytical Chemistry, Issue 89/11, 2017, Page(s) 5991-5997, ISSN 0003-2700
DOI: 10.1021/acs.analchem.7b00554

SET-LRP in biphasic mixtures of fluorinated alcohols with water
Author(s): Adrian Moreno, Tong Liu, Liang Ding, Irene Buzzacchera, Marina Galià, Martin Möller, Christopher J. Wilson, Gerard Lligadas, Virgil Percec
Published in: Polymer Chemistry, Issue 9/17, 2018, Page(s) 2313-2327, ISSN 1759-9954
DOI: 10.1039/c8py00062j

Exploring functional pairing between surface glycoconjugates and human galectins using programmable glycodendrimersomes
Author(s): Qi Xiao, Anna-Kristin Ludwig, Cecilia Romanò, Irene Buzzacchera, Samuel E. Sherman, Maria Vetro, Sabine Vértesy, Herbert Kaltner, Ellen H. Reed, Martin Möller, Christopher J. Wilson, Daniel A. Hammer, Stefan Oscarson, Michael L. Klein, Hans-Joachim Gabius, Virgil Percec
Published in: Proceedings of the National Academy of Sciences, Issue 115/11, 2018, Page(s) E2509-E2518, ISSN 0027-8424
DOI: 10.1073/pnas.1720055115

Cartilage Regeneration in Preannealed Silk Elastin-Like Co-Recombinamers Injectable Hydrogel Embedded with Mature Chondrocytes in an Ex Vivo Culture Platform
Author(s): Filippo Cipriani, Melanie Krüger, Israel Gonzalez de Torre, Luis Quintanilla Sierra, Matilde Alonso Rodrigo, Linda Kock, José Carlos Rodriguez-Cabello
Published in: Biomacromolecules, Issue 19/11, 2018, Page(s) 4333-4347, ISSN 1525-7797
DOI: 10.1021/acs.biomac.8b01211

Bicyclic RGD peptides with high integrin αvβ3 and α5β1 affinity promote cell adhesion on elastin-like recombinamers
Author(s): Filippo Cipriani, Dominik Bernhagen, Carmen Garcia-Arévalo, Israel González de Torre, Peter Timmerman, Jose Carlos Rodriguez-Cabello
Published in: Biomedical Materials, 2019, ISSN 1748-6041
DOI: 10.1088/1748-605x/aafd83

Sensor Devices Inspired by the Five Senses: A Review
Author(s): Mila I. Svechtarova, Irene Buzzacchera, B. Jelle Toebes, Jan Lauko, Nicoleta Anton, Christopher J. Wilson
Published in: Electroanalysis, 2016, ISSN 1040-0397
DOI: 10.1002/elan.201600047

Biomimetic Cell-Laden MeHA Hydrogels for the Regeneration of Cartilage Tissue
Author(s): Evgenia Tsanaktsidou, Olga Kammona, Norina Labude, Sabine Neuss, Melanie Krüger, Linda Kock, Costas Kiparissides
Published in: Polymers 2020, 12(7), 1598
DOI: 10.3390/polym12071598

Cellulose Nanofibril Hydrogel Promotes Hepatic Differentiation of Human Liver Organoids
Author(s): Melanie Krüger, Loes A. Oosterhoff, Monique E. van Wolferen, Simon A. Schiele, Andreas Walther, Niels Geijsen, Laura De Laporte, Luc J. W. van der Laan, Linda M. Kock, Bart Spee
Published in: Advanced Healthcare Materials 2020 Volume9, Issue6, 1901658
DOI: 10.1002/adhm.201901658

Anisometric Microstructures to Determine Minimal Critical Physical Cues Required for Neurite Alignment
Author(s): Sitara Vedaraman, Amaury Perez-Tirado, Tamas Haraszti, Jose Gerardo-Nava, Akihiro Nishiguchi, Laura De Laporte
Published in: Advanced Healthcare Materials 2021 Volume10, Issue20, 1901658
DOI: 10.1002/adhm.202100874

Bicyclic RGD peptides enhance nerve growth in synthetic PEG-based Anisogels
Author(s): Sitara Vedaraman, Dominik Bernhagen, Tamas Haraszti, Christopher Licht, Arturo Castro Nava, Abdolrahman Omidinia Anarkoli, Peter Timmerman, Laura De Laporte
Published in: Biomaterials Science (2021)
DOI: 10.1039/D0BM02051F

A Layer-by-Layer Single-Cell Coating Technique To Produce Injectable Beating Mini Heart Tissues via Microfluidics
Author(s): Luis P. B. Guerzoni, Yoshinari Tsukamoto, David B. Gehlen, Dirk Rommel, Tamás Haraszti, Mitsuru Akashi, Laura De Laporte
Published in: Biomacromolecules (2019)
DOI: 10.1021/acs.biomac.9b00786

Cellular responses to beating hydrogels to investigate mechanotransduction
Author(s): Yashoda Chandorkar, Arturo Castro Nava, Sjören Schweizerhof, Marcel Van Dongen, Tamás Haraszti, Jens Köhler, Hang Zhang, Reinhard Windoffer, Ahmed Mourran, Martin Möller, Laura De Laporte
Published in: Nature Communications (2019)
DOI: 10.1038/s41467-019-11475-4

Cell Encapsulation in Soft, Anisometric Poly(ethylene) Glycol Microgels Using a Novel Radical-Free Microfluidic System
Author(s): Luis P. B. Guerzoni, Jonas C. Rose, David B. Gehlen, Alexander Jans, Tamàs Haraszti, Matthias Wessling, Alexander J. C. Kuehne, Laura De Laporte
Published in: Small (2019)
DOI: 10.1002/smll.201900692

Nanofibrillar Cellulose as an Enzymatically and Flow Driven Degradable Scaffold for Three-Dimensional Tissue Engineering
Author(s): Melanie Krüger, Bart Spee, Andreas Walther, Laura De Laporte, Linda M. Kock
Published in: ASME. ASME J of Medical Diagnostics (2019)
DOI: doi.org/10.1115/1.4044473

Fibrous Hydrogels under Multi-Axial Deformation: Persistence Length as the Main Determinant of Compression Softening
Author(s): Paula de Almeida, Paul A. Janmey, Paul H. J. Kouwer
Published in: Advanced Functional Materials (2021)
DOI: 10.1002/adfm.202010527

Cell-matrix reciprocity in 3D culture models with nonlinear elasticity
Author(s): Kaizheng Liu, Maury Wiendels, Hongbo Yuan, Changshun Ruan, Paul H.J. Kouwer
Published in: Bioactive Materials (2022)
DOI: 10.1016/j.bioactmat.2021.08.002

Thin-Film Polyisocyanide-Based Hydrogels for Affinity Biosensors
Author(s): Daria Kotlarek, Kaizheng Liu, Nestor G. Quilis, Dominik Bernhagen, Peter Timmerman, Paul Kouwer, Jakub Dostalek
Published in: The Journal of Physical Chemistry C (2021)
DOI: doi.org/10.1021/acs.jpcc.1c02489

Actuated plasmonic nanohole arrays for sensing and optical spectroscopy applications
Author(s): Daria Kotlarek, Stefan Fossati,Priyamvada Venugopalan, Nestor Gisbert Quilis, Jiří Slabý, Jiří Homola, Médéric Lequeux, Frédéric Amiard,Marc Lamy de la Chapelle, Ulrich Jonas, Jakub Dostálek
Published in: Nanoscale (2020)
DOI: 10.1039/D0NR00761G

Compact Grating-Coupled Biosensor for the Analysis of Thrombin
Daria Kotlarek, Mariia Vorobii, Wojciech Ogieglo, Wolfgang Knoll, Cesar Rodriguez-Emmenegger, Jakub Dostálek
Published in: ACS Sensors (2019)
DOI: 10.1021/acssensors.9b00827

Actively Tunable Collective Localized Surface Plasmons by Responsive Hydrogel Membrane
Nestor Gisbert Quilis, Marcel van Dongen, Priyamvada Venugopalan, Daria Kotlarek, Christian Petri, Alberto Moreno Cencerrado, Sorin Stanescu, Jose Luis Toca Herrera, Ulrich Jonas, Martin Möller, Ahmed Mourran, Jakub Dostalek
Published in: Advanced Optical Materials (2021)
DOI: 10.1002/adfm.202010527

Encapsulation of hydrophobic components in dendrimersomes and decoration of their surface with proteins and nucleic acids
Paola Torre, Qi Xiao, Irene Buzzacchera, Samuel E. Sherman, Khosrow Rahimi, Nina Yu. Kostina, Cesar Rodriguez-Emmenegger, Martin Möller, Christopher J. Wilson, Michael L. Klein, Matthew C. Good, Virgil Percec
Published in: Proceedings of the National Academy of Sciences (PNAS, 2019)
DOI: doi.org/10.1073/pnas.1904868116

Hydrophobic Cholesteryl Moieties Trigger Substrate Cell–Membrane Interaction of Elastin–Mimetic Protein Coatings in Vitro
Leander Poocza, Filippo Cipriani, Matilde Alonso, José Carlos Rodríguez-Cabello
Published in: ACS Omega (2019)
DOI: 10.1021/acsomega.9b00548

Charge Density as a Molecular Modulator of Nanostructuration in Intrinsically Disordered Protein Polymers
Sergio Acosta, Leander Poocza, Luis Quintanilla-Sierra, José Carlos Rodríguez-Cabello
Published in: Biomacromolecules (2020)
DOI: 10.1021/acs.biomac.0c00934

Structural characterization of fibrous synthetic hydrogels using fluorescence microscopy
Johannes Vandaele, Boris Louis, Kaizheng Liu, Rafael Camacho, Paul H. J. Kouwer, Susana Rocha
Published in: Soft Matter (2020)
DOI: 10.1039/C9SM01828J

Synthetic Extracellular Matrices as a Toolbox to Tune Stem Cell Secretome
Kaizheng Liu, Tomas Veenendaal, Maury Wiendels, Alejandra M. Ruiz-Zapata, Justin van Laar, Rafail Kyranas, Hilde Enting, Bram van Cranenbroek, Hans J. P. M. Koenen, Silvia M. Mihaila, Egbert Oosterwijk, Paul H. J. Kouwer
Published in: ACS Applied Materials and Interfaces (2020)
DOI: 10.1021/acsami.0c16208

Responsive polymer networks and brushes for active plasmonics
N. G. Quilis, N. Sharma, S. Fossati, W. Knoll, J. Dostalek
Published in: ACS Applied Materials and Interfaces (2017)
DOI: 10.1002/9781119455042.ch24

Newsletters
Bi-annual newsletters to announce training events and scientific results

14 (anticipated) PhD diplomas
Awarding (anticipated) doctoral degrees by all partners

Bicyclic peptides
Different improved biological bicyclic peptides with integrin binding domains and proteolytic cleavage sites

Optical characterization methods
Characterization of thin film hydrogels by optical waveguide spectroscopy (OWS) and surface plasmon resonance (SPR)

Completion of Recruitment