The Immobilization of Hyaluronic Acid in 3D Hydrogel Scaffolds Modulates Macrophage Polarization

Abstract

Macrophages are key modulators of immunity, tissue homeostasis and disease development. As our understanding of macrophage biology and their tissue specific behaviors grow the necessity to model macrophages within a 3D biomimetic environment becomes increasingly apparent. Numerous hydrogels have been developed and explored for this purpose, extracellular matrix (ECM) mimicking hydrogels gaining a special interest. In this study, we present the use of such a hydrogel composed of collagen and hyaluronic acid (HA), two of the major ECM components, for the 3D culture of macrophages to model their role in different tissues and diseases. We demonstrate the ability to tailor the mechanical properties of the hydrogel through formulation modulation. Human macrophages retain morphology, viability, and expression of key cell surface markers when 3D cultured within the hydrogel. Interestingly, we demonstrate in this work, that independent of mechanical properties, by adjusting the composition of the hydrogel, specifically HA molecular weight, we can steer macrophage polarization towards either a pro-inflammatory or anti-inflammatory phenotype. This HA-dependent modulation of macrophage behavior is nullified if the HA is chemically crosslinked, shedding light on the impact of one of the most commonly used preparation methods for collagen-HA hydrogels on macrophage behavior. Graphical abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=84 SRC="FIGDIR/small/588451v1_ufig1.gif" ALT="Figure 1"> View larger version (20K): org.highwire.dtl.DTLVardef@1cbe667org.highwire.dtl.DTLVardef@1837e58org.highwire.dtl.DTLVardef@1dbdbc6org.highwire.dtl.DTLVardef@139d65b_HPS_FORMAT_FIGEXP M_FIG C_FIG Hyaluronic acid (HA) is an extracellular matrix component, which can modulate the polarization of macrophages. The chemical crosslinking of HA to hydrogel scaffolds counteracts the cell signaling role of this molecule, preventing HA from modulating the immune polarization of macrophages within the model. Key pointsO_LICollagen-HA hydrogels form suitable biomimetic niches for the 3D culture of human macrophages in which hydrogel composition modulates macrophage phenotype, specifically the presence and molecular weight of HA. C_LIO_LIPolarization of macrophage phenotype by HA is nullified if HA is chemically cross-linked within the hydrogel. C_LI