Patterned alginate hydrogel spatially guides collagen fibrillogenesis, viscoelasticity and endothelial cell invasion

bioRxiv (Cold Spring Harbor Laboratory)

Abstract

Abstract Angiogenesis following injury has been shown to be driven by fibrillar proteins of the extracellular matrix (ECM), such as collagen. However, the use of protein-based biomaterials presents some challenges, such as uncontrolled degradation and limited tuneability. We demonstrate how to create patterned interpenetrating networks (IPNs) based on covalently crosslinked alginate and physically crosslinked collagen that provide suitable mechanical properties to support migration of endothelial cells (ECs) in a spatially controlled manner. Low molecular weight alginate is functionalized with norbornene (N) or tetrazine (T), which enables two independent covalent crosslinking methods: UV-mediated and degradable crosslinks with matrix metalloproteinase (MMP) sensitive peptides (Deg) and slower spontaneous N:T non-degradable crosslinks (noDeg). Using photolithography, patterns in degradation, collagen fibrillogenesis, microarchitecture and matrix viscoelasticity are created. The potential of such 3D patterned alginate-collagen (Alg-Col) IPNs to spatially guide EC invasion and proliferation was tested in a microfluidics platform resembling an early healing setting. Only regions combining collagen fibrillogenesis, alginate degradability and viscoelasticity demonstrated EC cell invasion similar to the ones found in vivo following injury. The 3D patterned Alg-Col IPNs are compatible with microfluidics, offer an strategy to widen the applications of protein-based hydrogels and present a versatile platform for tissue engineering and disease modeling. Table of Contents 3D patterned alginate–collagen interpenetrating networks spatially control collagen fibrillogenesis, matrix degradability, microarchitecture, and viscoelasticity. By combining orthogonal crosslinking with photolithographic patterning, distinct matrix regions guide endothelial cell invasion and proliferation. These hybrid materials recapitulate key angiogenic responses following injury and can be readily incorporated into microfluidic chips, providing a versatile platform for tissue engineering and disease modeling.

Authors 9

1. Claudia A. Garrido Aachen DWI−Leibniz Institute for Interactive Materials Aachen

   Max Planck Institute of Colloids and Interfaces · DWI – Leibniz Institute for Interactive Materials · Berlin Institute of Health at Charité - Universitätsmedizin Berlin · Charité - Universitätsmedizin Berlin

   Affiliation as printed

   DWI – Leibniz-Institute for Interactive Materials. Aachen, Germany

   Julius Wolff Institute for Biomechanics and Musculoskeletal Regeneration, Berlin Institute for Health at Charité - Universitätsmedizin Berlin, Berlin, Germany

   Max Planck Institute for Colloids and Interfaces, Potsdam, Germany

2. Daniela S. Garske

   Max Planck Institute of Colloids and Interfaces · Berlin Institute of Health at Charité - Universitätsmedizin Berlin · Charité - Universitätsmedizin Berlin

   Affiliation as printed

   Julius Wolff Institute for Biomechanics and Musculoskeletal Regeneration, Berlin Institute for Health at Charité - Universitätsmedizin Berlin, Berlin, Germany

   Max Planck Institute for Colloids and Interfaces, Potsdam, Germany

3. Bernhard Häßel Aachen DWI−Leibniz Institute for Interactive Materials Aachen Institute of Technical and Macromolecular Chemistry Chair for Macromolecular Materials for Medicine

   DWI – Leibniz Institute for Interactive Materials · RWTH Aachen University

   Affiliation as printed

   DWI – Leibniz-Institute for Interactive Materials. Aachen, Germany

   Institute of Technical and Macromolecular Chemistry (ITMC), Chair for Macromolecular Materials for Medicine, RWTH University Aachen, Aachen, Germany

4. C Bastard Aachen DWI−Leibniz Institute for Interactive Materials Aachen Institute of Technical and Macromolecular Chemistry Chair for Macromolecular Materials for Medicine

   DWI – Leibniz Institute for Interactive Materials · RWTH Aachen University

   Affiliation as printed

   DWI – Leibniz-Institute for Interactive Materials. Aachen, Germany

   Institute of Technical and Macromolecular Chemistry (ITMC), Chair for Macromolecular Materials for Medicine, RWTH University Aachen, Aachen, Germany

5. Julia van de Kamp Aachen DWI−Leibniz Institute for Interactive Materials Aachen Institute of Technical and Macromolecular Chemistry Chair for Macromolecular Materials for Medicine

   DWI – Leibniz Institute for Interactive Materials · RWTH Aachen University

   Affiliation as printed

   DWI – Leibniz-Institute for Interactive Materials. Aachen, Germany

   Institute of Technical and Macromolecular Chemistry (ITMC), Chair for Macromolecular Materials for Medicine, RWTH University Aachen, Aachen, Germany

6. Laura De Laporte Aachen DWI−Leibniz Institute for Interactive Materials Aachen Institute for Applied Medical Engineering Institute of Technical and Macromolecular Chemistry Chair for Macromolecular Materials for Medicine

   DWI – Leibniz Institute for Interactive Materials · Universitätsklinikum Aachen · RWTH Aachen University

   Affiliation as printed

   DWI – Leibniz-Institute for Interactive Materials. Aachen, Germany

   Institute for Applied Medical Engineering (AME), Advanced Materials for Biomedicine (AMB), University Hospital RWTH Aachen, Institute of Technical and Macromolecular Chemistry (ITMC), Center for Biohybrid Medical Systems (CMBS), Aachen, Germany

   Institute of Technical and Macromolecular Chemistry (ITMC), Chair for Macromolecular Materials for Medicine, RWTH University Aachen, Aachen, Germany

7. Georg N. Duda

   Berlin Institute of Health at Charité - Universitätsmedizin Berlin · Charité - Universitätsmedizin Berlin

   Affiliation as printed

   Julius Wolff Institute for Biomechanics and Musculoskeletal Regeneration, Berlin Institute for Health at Charité - Universitätsmedizin Berlin, Berlin, Germany

8. Katharina Schmidt‐Bleek

   Berlin Institute of Health at Charité - Universitätsmedizin Berlin · Charité - Universitätsmedizin Berlin

   Affiliation as printed

   Julius Wolff Institute for Biomechanics and Musculoskeletal Regeneration, Berlin Institute for Health at Charité - Universitätsmedizin Berlin, Berlin, Germany

9. Amaia Cipitria corresponding

   Ikerbasque · Biogipuzkoa Health Research Institute

   Affiliation as printed

   Group of Bioengineering in Regeneration and Cancer, Biogipuzkoa Health Research Institute, San Sebastián, Spain

   IKERBASQUE, Basque Foundation for Science, Bilbao, Spain