Effect of molecular weight of tyramine-modified hyaluronan on polarization state of THP-1 and peripheral blood mononuclear cells-derived macrophages

Abstract

The immunomodulatory properties of hyaluronan and its derivatives are key to their use in medicine and tissue engineering. In this work we evaluated the capability of soluble tyramine-modified hyaluronan (THA) of two molecular weights (low Mw=280 kDa and high MMw=1640 kDa) for polarization of peripheral blood mononuclear cells-derived macrophages. We demonstrate the polarization effects of the supplemented THA by the semi-automated image analysis from confocal microscopy, immunofluorescent staining utilising CD68 and CD206 surface markers, RT-qPCR gene expression analysis, as well as using the enzyme-linked immunosorbent assay (ELISA). The expression of certain cytokines including TNF-α (pro-inflammatory), IL-10 (anti-inflammatory) and surface marker CD206 are more viable options for rapid screening of polarization states of macrophages with CD206 marker being associated with the lowest donor variability. Our findings indicate that low MwTHA can be associated with mild pro-inflammatory state, M1-like state, whereas high MwTHA can be associated with mild pro-regenerative, M2-like state, and that the extent of these responses are donor-dependent. The variation in the obtained phenotypic responses may stem from possible uncontrolled genetic factors in received blood donor samples. Finally, we confirm the typical association of low versus high molecular weight for THA holds true and stipulate that these responses will provide more accurate in vivo representation and translational immunomodulatory guidance for the use of THA-based biomaterials to a wider biomaterials and tissue engineering communities.