Iron nanoparticle-labeled murine mesenchymal stromal cells in an osteoarthritic model persists and suggests anti-inflammatory mechanism of action

Amanda M Hamilton; Wing-Yee Cheung; Alejandro Gómez-Aristizábal; Anirudh Sharma; Sayaka Nakamura; Amélie Chaboureau; Shashank Bhatt; Razieh Rabani; Mohit Kapoor; Paula J Foster; Sowmya Viswanathan

doi:10.1371/journal.pone.0214107

Iron nanoparticle-labeled murine mesenchymal stromal cells in an osteoarthritic model persists and suggests anti-inflammatory mechanism of action

PLoS One. 2019 Dec 3;14(12):e0214107. doi: 10.1371/journal.pone.0214107. eCollection 2019.

Authors

Amanda M Hamilton¹, Wing-Yee Cheung², Alejandro Gómez-Aristizábal², Anirudh Sharma², Sayaka Nakamura², Amélie Chaboureau², Shashank Bhatt², Razieh Rabani^{2

3}, Mohit Kapoor^{2

3

4}, Paula J Foster^{1

5}, Sowmya Viswanathan^{2

3

6

7}

Affiliations

¹ Imaging Research Laboratories, Robarts Research Institute, London, ON, Canada.
² The Arthritis Program, Toronto Western Hospital, Toronto, ON, Canada.
³ Division of Genetics and Development, Krembil Research Institute, University Health Network, Toronto, ON, Canada.
⁴ Department of Surgery, Department of Laboratory Medicine and Pathobiology, University of Toronto, Toronto, ON, Canada.
⁵ Department of Medical Biophysics, Schulich School of Medicine and Dentistry, Western University, London, ON, Canada.
⁶ Institute of Biomaterials and Biomedical Engineering, University of Toronto, Toronto, ON, Canada.
⁷ Department of Medicine, University of Toronto, Toronto, ON, Canada.

Abstract

Osteoarthritis (OA) is characterized by cartilage degradation and chronic joint inflammation. Mesenchymal stem cells (MSCs) have shown promising results in OA, but their mechanism of action is not fully understood. We hypothesize that MSCs polarize macrophages, which are strongly associated with joint inflammation to more homeostatic sub-types. We tracked ferumoxytol (Feraheme™, iron oxide nanoparticle)-labeled murine MSCs (Fe-MSCs) in murine OA joints, and quantified changes to joint inflammation and fibrosis. 10-week-old C57BL/6 male mice (n = 5/group) were induced to undergo osteoarthritis by destabilization of medical meniscus (DMM) or sham surgery. 3 weeks post-surgery, mice were injected intra-articularly with either fluorescent dye-(DiR) labeled or DiR-Fe-MSC or saline to yield 4 groups (n = 5 per group for each timepoint [1, 2 and 4weeks]). 4 weeks after injection, mice were imaged by MRI, and scored for i) OARSI (Osteoarthritis Research Society International) to determine cartilage damage; ii) immunohistochemical changes in iNOS, CD206, F4/80 and Prussian Blue/Sca-1 to detect pro-inflammatory, homeostatic and total macrophages and ferumoxytol -labeled MSCs respectively, and iii) Masson's Trichrome to detect changes in fibrosis. Ferumoxytol-labeled MSCs persisted at greater levels in DMM vs. SHAM-knee joints. We observed no difference in OARSI scores between MSC and vehicle groups. Sca-1 and Prussian Blue co-staining confirmed the ferumoxytol label resides in MSCs, although some ferumoxytol label was detected in proximity to MSCs in macrophages, likely due to phagocytosis of apoptotic MSCs, increasing functionality of these macrophages through MSC efferocytosis. MRI hypertintensity scores related to fluid edema decreased in MSC-treated vs. control animals. For the first time, we show that MSC-treated mice had increased ratios of %CD206+: %F4/80+ (homeostatic macrophages) (p<0.05), and decreased ratios of %iNOS+: %F4/80+ macrophages (p<0.01), supporting our hypothesis that MSCs may modulate synovial inflammation.

Publication types

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

MeSH terms

Animals
Anti-Inflammatory Agents / pharmacology
Cartilage, Articular
Disease Models, Animal
Ferric Compounds
Humans
Inflammation / drug therapy
Injections, Intra-Articular
Iron / metabolism
Iron / therapeutic use
Knee Joint
Male
Mesenchymal Stem Cell Transplantation / methods
Mesenchymal Stem Cells / physiology*
Metal Nanoparticles / therapeutic use*
Mice
Mice, Inbred C57BL
Nanoparticles
Osteoarthritis / therapy*

Substances

Anti-Inflammatory Agents
Ferric Compounds
ferric oxide
Iron

Grants and funding

This study received peer review funding from the Ontario Institute for Regenerative Medicine and Stem Cell Network and funding from The Arthritis Society (15-321) and Stem Cell Network (Clinical Trial Impact grant # 8). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.