In vitro evaluation of iron oxide nanoparticle-induced thromboinflammatory response using a combined human whole blood and endothelial cell model

Alexandra Gerogianni; Melissa Bal; Camilla Mohlin; Trent M Woodruff; John D Lambris; Tom E Mollnes; Dick J Sjöström; Per H Nilsson

doi:10.3389/fimmu.2023.1101387

In vitro evaluation of iron oxide nanoparticle-induced thromboinflammatory response using a combined human whole blood and endothelial cell model

Front Immunol. 2023 Apr 4:14:1101387. doi: 10.3389/fimmu.2023.1101387. eCollection 2023.

Authors

Alexandra Gerogianni^{1

2}, Melissa Bal^{1

2}, Camilla Mohlin^{1

2}, Trent M Woodruff³, John D Lambris⁴, Tom E Mollnes^{5

6

7}, Dick J Sjöström^{1

2}, Per H Nilsson^{1

2

5}

Affiliations

¹ Linnaeus Centre for Biomaterials Chemistry, Linnaeus University, Kalmar, Sweden.
² Department of Chemistry and Biomedicine, Linnaeus University, Kalmar, Sweden.
³ School of Biomedical Sciences, Faculty of Medicine, The University of Queensland, Brisbane, QLD, Australia.
⁴ Department of Pathology and Laboratory Medicine, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, United States.
⁵ Department of Immunology, Oslo University Hospital and University of Oslo, Oslo, Norway.
⁶ Centre of Molecular Inflammation Research, Department of Clinical and Molecular Medicine, Norwegian University of Science and Technology, Trondheim, Norway.
⁷ Research Laboratory, Nordland Hospital, Bodo, Norway.

Abstract

Iron oxide nanoparticles (IONPs) are widely used in diagnostic and therapeutic settings. Upon systemic administration, however, they are rapidly recognized by components of innate immunity, which limit their therapeutic capacity and can potentially lead to adverse side effects. IONPs were previously found to induce the inflammatory response in human whole blood, including activation of the complement system and increased secretion of cytokines. Here, we investigated the thromboinflammatory response of 10-30 nm IONPs in lepirudin anticoagulated whole blood in interplay with endothelial cells and evaluated the therapeutic effect of applying complement inhibitors to limit adverse effects related to thromboinflammation. We found that IONPs induced complement activation, primarily at the C3-level, in whole blood incubated for up to four hours at 37°C with and without human microvascular endothelial cells. Furthermore, IONPs mediated a strong thromboinflammatory response, as seen by the significantly increased release of 21 of the 27 analyzed cytokines (p<0.05). IONPs also significantly increased cell-activation markers of endothelial cells [ICAM-1 (p<0.0001), P/E-selectin (p<0.05)], monocytes, and granulocytes [CD11b (p<0.001)], and platelets [CD62P (p<0.05), CD63 (p<0.05), NAP-2 (p<0.01), PF4 (p<0.05)], and showed cytotoxic effects, as seen by increased LDH (p<0.001) and heme (p<0.0001) levels. We found that inflammation and endothelial cell activation were partly complement-dependent and inhibition of complement at the level of C3 by compstatin Cp40 significantly attenuated expression of ICAM-1 (p<0.01) and selectins (p<0.05). We show that complement activation plays an important role in the IONPs-induced thromboinflammatory response and that complement inhibition is promising in improving IONPs biocompatibility.

Keywords: complement activation; complement inhibition; endothelial cells; iron oxide nanoparticles; thromboinflammation; whole blood.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Complement System Proteins / metabolism
Cytokines / metabolism
Endothelial Cells* / metabolism
Humans
Inflammation / metabolism
Intercellular Adhesion Molecule-1 / metabolism
Magnetic Iron Oxide Nanoparticles
Thrombosis* / drug therapy
Thrombosis* / metabolism

Substances

Intercellular Adhesion Molecule-1
Complement System Proteins
Cytokines

Grants and funding

This work was supported by the Norwegian Research Council (Project 274332), the Swedish Research Council (Project 2018-04087), and the Crafoord Foundation (Project 20210961 and 20190890).