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Gelatin-Hyaluronan Click-Crosslinked Cryogels Elucidate Human Macrophage Invasion Behavior |
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| Authors: | Laura C Bahlmann Alexander E G Baker Chang Xue Sophie Liu Moritz Meier-Merziger Danielle Karakas Luke Zhu Ileana Co Spencer Zhao Allysia Chin Alison McGuigan John Kuruvilla Rob C Laister Molly S Shoichet |
| Affiliation: | 1. Institute of Biomedical Engineering, 164 College Street, Toronto, Ontario, M5S 3G9 Canada
The Donnelly Centre, University of Toronto, 160 College St, Toronto, Ontario, M5S 3E1 Canada;2. Institute of Biomedical Engineering, 164 College Street, Toronto, Ontario, M5S 3G9 Canada The Donnelly Centre, University of Toronto, 160 College St, Toronto, Ontario, M5S 3E1 Canada Department of Chemical Engineering and Applied Chemistry, University of Toronto, 200 College Street, Toronto, Ontario, M5S 3E5 Canada;3. Department of Chemical Engineering and Applied Chemistry, University of Toronto, 200 College Street, Toronto, Ontario, M5S 3E5 Canada;4. Department of Chemistry, Johannes Gutenberg University, Duesbergweg 10-14, 55128 Mainz, Germany;5. Institute of Biomedical Engineering, 164 College Street, Toronto, Ontario, M5S 3G9 Canada;6. Institute of Biomedical Engineering, 164 College Street, Toronto, Ontario, M5S 3G9 Canada Department of Chemical Engineering and Applied Chemistry, University of Toronto, 200 College Street, Toronto, Ontario, M5S 3E5 Canada;7. Princess Margaret Cancer Centre, University Health Network, 610 University Ave, Toronto, Ontario, M5G 2C1 Canada |
| Abstract: | Hydrogel models of metastasis traditionally focus on the invasion of cancer cells; however, other cells in the tumor microenvironment that are associated with metastasis also have the ability to migrate. Macrophage phenotype plays a key role in the tumor microenvironment, yet understanding their migration within tunable 3D in vitro models has been limited. To gain a greater understanding of macrophage invasive behavior, stable and transparent oxime-crosslinked cryogels comprised of click-crosslinked gelatin-oxyamine and hyaluronan-aldehyde (GELox-HAa) are synthesized. Fibronectin-derived, oxyamine-modified PHSRN-RGDSP peptides are incorporated to further mimic the tumor extracellular matrix without impacting cryogel mechanics. It is found that primary human macrophages migrate to a greater depth in cryogels with greater porosity and pore size. To better understand the mechanism of migration, cells are treated with either inhibitors of matrix metalloproteinases (MMPs) or rho-associated protein kinase (ROCK) and a predominantly MMP-mediated mechanism of invasion is found. Macrophage polarization studies reveal that anti-inflammatory, interleukin-4/13 (IL4/IL13)-treated macrophages migrate through cryogels to a greater extent than pro-inflammatory, interferon-gamma/lipopolysaccharide (IFNγ/LPS)-treated cells. Interestingly, polarized macrophages move through cryogels using a combination of amoeboid and mesenchymal migration. These findings of macrophage invasion in this cryogel platform set the stage for their further study in a biomimetic tumor microenvironment. |
| Keywords: | cryogel extracellular matrix gelatin hyaluronan macrophage invasion |