Glycosaminoglycan-functionalized hydrogels for sustained delivery of tissue inhibitor of metalloproteinase-3 mediating matrix metalloprotease inhibition and extracellular matrix stabilization
Excessive protease activity and impaired tissue regeneration are hallmarks of many disease states. Elevated matrix metalloproteinase-9 (MMP-9) plays a key role in adverse tissue remodeling by excessively degrading extracellular matrix (ECM) components and growth factors. Tissue inhibitor of metalloproteinase-3 (TIMP-3) regulates ECM turnover, and its bioavailability is influenced by glycosaminoglycans (GAGs). This study aimed to develop a methacrylated gelatin (GelMA)-based hydrogel functionalized with acrylated sulfated hyaluronan (sHA<sub>c</sub>) as a TIMP-3 delivery system to decrease ECM degradation under pathophysiological conditions. sHA<sub>c</sub> incorporation enhanced hydrogel stiffness, reduced degradation rates and yielded sustained TIMP-3 release for up to 28 days. Molecular modeling and surface plasmon resonance demonstrated preferential binding of TIMP-3 to sHA<sub>c</sub> over hyaluronan methacrylates, together providing a molecular rationale for the reduced and sustained release of TIMP-3 from sHA<sub>c</sub>-containing hydrogels. Angiogenesis-related functional assays, supported by molecular modeling studies, indicate that sHA<sub>c</sub> modulates the anti-angiogenic activity of TIMP-3 by altering vascular endothelial growth factor receptor-associated signaling, while preserving metalloproteinase inhibition. Released TIMP-3 from GelMA/sHA<sub>c</sub> hydrogels retained bioactivity, effectively inhibiting MMP-9 activity and mitigating ECM degradation in-vitro and in human ex-vivo models. In a murine subcutaneous implantation model, sHA<sub>c</sub>-functionalized TIMP-3-loaded hydrogels were associated with reduced inflammatory cell presence and altered vascular- and matrix-related tissue signatures compared with GelMA controls. These findings underscore the potential of sHA<sub>c</sub>-functionalized GelMA hydrogels as biomaterials for therapeutics delivery, offering controlled TIMP-3 release and sustained bioactivity to promote ECM stability and on-demand MMP inhibition. This system represents a promising strategy for addressing the challenges of excessive MMP activity.
Authors
- Emmanuel Ampofo
- Poh Soo Lee (ORCID: https://orcid.org/0000-0003-2285-3621)
- Thomas Tschernig (ORCID: https://orcid.org/0000-0002-7788-1796)
- Letícia Prates Roma (ORCID: https://orcid.org/0000-0001-9527-0529)
- Matthias W. Laschke (ORCID: https://orcid.org/0000-0002-7847-8456)
- Matthias Hannig (ORCID: https://orcid.org/0000-0003-0669-6881)
- Marcel A. Lauterbach (ORCID: https://orcid.org/0009-0008-8145-6873)
- Gloria Ruiz‐Gómez
- Mathias Hohl (ORCID: https://orcid.org/0000-0001-6946-9825)
- Stefan Rupf (ORCID: https://orcid.org/0000-0002-1551-9935)
- Albrecht Berg
- Selina Wrublewsky
- Charlotte Berhorst
- Cedric Wilden
- Therese Steudter
- Stephanie Moeller
- Sandra Rother
- Tobias A. Dancker
- M. Teresa Pisabarro
- Dalia Alansary
- Markus Bischoff
- Ben Wieland
- Bergita Ganse
- Wolfgang Metzger
- Caroline Bickelmann
- Paula Marie Schindler
- Fabian Junker
Institutions
- Leibniz-Institute for New Materials (DE)
- Innovent (DE)
- Max Bergmann Zentrum für Biomaterialien (DE)
- Institute of Medical Microbiology and Hygiene (DE)
- Healing Foundation (AU)
- Technische Universität Dresden (DE)
- Saarland University (DE)
Publication Details
- Journal
- Bioactive Materials
- Published
- 2026-02-12
- DOI
- https://doi.org/10.1016/j.bioactmat.2026.02.010
- Primary Topic
- Protease and Inhibitor Mechanisms
- Type
- article
- Field-Weighted Citation Impact
- 0.00