Effects of WF10 on Glycosaminoglycan Sulphation in Proinflammatory Monocytes and Macrophages

The chlorite-based drug solution WF10 has been successfully applied to dampen strong inflammatory disease states and to improve wound healing processes. Howev‐ er, the molecular mechanisms of this drug are not well understood. This study is di‐ rected to investigate how WF10 and its components affect the expression of surface markers and sulphated proteoglycans and glycosaminoglycans in proinflammatorystimulated monocytes and macrophages. Human blood-derived macrophages were cultivated from monocytes in the presence of 50 U/ml granulocyte-macrophage colony-stimulating factor and activated by a mix‐ ture of 100 ng/ml lipopolysaccharide (LPS) and 10 ng/ml interferon γ (IFNγ). These cells were identified and characterised by their specific cell-surface receptors CD14, CD16, CD80, CD86, CD163, and CD206 using flow cytometry approaches. The sulpha‐ tion level of proteoglycans and glycosaminoglycans was assessed by the BlyscanTM dye-binding assay. The expression of the surface marker CD44, a proteoglycan with sulphated glycosaminoglycan side chains, was followed by antibodies against CD44. The binding of fluorescence-labelled hyaluronan to CD44 was also investigated by flow cytometry. All analyses were performed after incubation of monocytes and mac‐ rophages with WF10 or with its main components chlorite and chlorate. The drug substance WF10 inhibited the activation of LPS/IFNγ-stimulated human monocyte-derived macrophages. Among them are the diminished expression of proinflammatory surface markers, the inhibition of the expression of the hyaluronan receptor CD44, and the binding of hyaluronan to CD44. Further, the overall amount of sulphated proteoglycans and glycosaminoglycans was down-regulated by WF10. These in vitro experiments indicate that WF10 is able to inhibit the proinflammatory activation of macrophages. The results suggested that chlorite is the active principle in WF10 as chlorite caused principally the same changes in targets as WF10. The WF10 component chlorate inhibited only the overall sulphation level of proteoglycans and glycosaminoglycans and the binding of hyaluronan to CD44. © 2016 The Author(s). Licensee InTech. This chapter is distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/3.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. To sum up, WF10 is a promising tool to inhibit proinflammatory states of immune cells. The inhibition of activation processes in monocytes and macrophages by WF10 coincides well with the results about clinical application of WF10.