Inducing healing-like human primary macrophage phenotypes by 3D hydrogel coated nanofibres

Immune cells are present in the blood and in resident tissues, and the nature of their reaction towards biomaterials is decisive for materials success or failure. Macrophages may for example be classically activated to trigger inflammation (M1), or alternatively activated which supports healing and vascularisation (M2). Here, we have generated 3D nanofibrous meshes in different porosities and precisely controlled surface chemistries comprising PLGA, hydrogel-coated protein repellant and protein repellant endowed with the bioactive peptide sequences GRGDS or GLF. We also prepared 2D substrates with corresponding surface chemistry for a systematic evaluation of primary human macrophage adhesion, migration, transcriptome expression, cytokine release and surface marker expression. Our data show that material morphology is a powerful means in biomaterial design to influence immune cell response. Flat substrates lead to an increased number of M2 classified CD163(+) macrophages. However, these M2 cells released large amounts of pro-inflammatory cytokines. In contrast, 3D nanofibres with corresponding surface chemistry yielded M1 classified 27E10(+) macrophages with a significantly increased release of pro-angiogenic chemokines and angiogenesis related molecules and a strong decrease of pro-inflammatory cytokines. We thus suggest that, for macrophages in contact with biomaterials, cytokine release is taken as main criterion instead of surface-markers for macrophage classifications.