SAT0004 Intra-articular injection of adipose-derived mesenchymal stromal cells reduce experimental oa pathology via il-1Β-mediated reallocation and enhanced phagocytosis of polymorphonuclear leucocytes
Authors: S. van Dalen, M. van den Bosch, A. Blom, P. van der Kraan, T. Vogl, L. Casteilla, P. van Lent
Journal: Annals of the Rheumatic Diseases (2018)

<h3>Background</h3> Injection of adipose-derived mesenchymal stromal cells (ASCs) into knee joints after induction of experimental inflammatory osteoarthritis (CiOA) reduces development of joint pathology.<sup>1</sup> This protection is only achieved when ASCs are applied in early CiOA, which is characterised by synovitis and high levels of S100A8/A9 and IL-1β, suggesting that inflammation boosts the protective effect of ASCs.<sup>2</sup> <h3>Objectives</h3> To examine the role of synovitis in ASC-mediated amelioration of CiOA pathology. <h3>Methods</h3> CiOA was induced by intra-articular collagenase injection. Knee joint sections were stained with haematoxylin/eosin and immunolocalization of polymorphonuclear leucocytes (PMNs) and ASCs was performed using antibodies for NIMP-R14 and CD271, respectively. Chemokine expression induced by IL-1β or S100A8/A9 was assessed with qPCR and Luminex. Migration of PMNs through transwell membranes towards ASC-conditioned medium (CM) was examined using flow cytometry. ASC-PMN co-cultures were analysed microscopically and with Luminex. Phagocytic capacity of PMNs was measured with labelled zymosan particles. <h3>Results</h3> Intra-articular injection of saline in knee joints of day 7 CiOA induced a flare already after 6 hours, characterised by particularly PMNs scattered throughout the synovium. Although ASC injection resulted in comparable numbers of PMNs, these cells however, were clustered around ASCs. IL-1β-stimulation of ASCs <i>in vitro</i> strongly increased expression of PMN-attracting chemokines KC, CXCL5, and CXCL7, whereas S100A8/A9 did not. Migration of PMNs towards CM of IL-1β-stimulated ASCs (IL-1β-CM) was significantly enhanced (2.9-fold increase) when compared to CM of non-stimulated ASCs (NS-CM). After 6 hours co-culturing PMNs with IL-1β-stimulated ASCs, the number of clustered PMNs per ASC was significantly increased. Interestingly, association of PMNs with ASCs significantly diminished the release of KC protein by ASCs (69% lower after 24 hour), and also strongly reduced the release of S100A8/A9 protein by the PMNs. Moreover, phagocytic capacity of PMNs was strongly enhanced after priming with CM of IL-1β-stimulated ASCs. <h3>Conclusions</h3> Local application of ASCs in inflamed CiOA knee joints results in attraction and clustering of PMNs with ASCs in the synovium, which is likely mediated by IL-1β-induced up-regulation of chemokine release by ASCs. This results in lowered S100A8/A9 levels and enhanced phagocytic capacity of PMNs, enabling the clearance of debris to attenuate synovitis and promote tissue repair. <h3>References</h3> [1] ter Huurne M, et al. Antiinflammatory and chondroprotective effects of intraarticular injection of adipose-derived stem cells in experimental osteoarthritis. Arthritis Rheum2012. Nov;64(11):3604–13. [2] Schelbergen RF, et al. Treatment efficacy of adipose-derived stem cells in experimental osteoarthritis is driven by high synovial activation and reflected by S100A8/A9 serum levels. Osteoarthritis Cartilage 2014 Aug;22(8):1158–66. <h3>Disclosure of Interest</h3> None declared