Adipose stromal cells mediated switching of the pro-inflammatory profile of M1-like macrophages is facilitated by PGE2: in vitro evaluation.
To define if adipose mesenchymal stromal cell (ASC) treatment mediated switching of the pro-inflammatory profile of M1-like macrophages as a means to develop a tailored in vitro efficacy/potency test.



We firstly performed immunohistochemical analysis of CD68, CD80 (M1-like) and CD206 (M2-like) macrophages in osteoarthritic (OA) synovial tissue. ASC were co-cultured in contact and in transwell with activated (GM-CSF + IFNγ)-M1 macrophages. We analyzed IL1β, TNFα, IL6, MIP1α/CCL3, S100A8, S100A9, IL10, CD163 and CD206 by qRT-PCR or immunoassays. Prostaglandin E2 (PGE2) blocking experiments were performed using PGE2 receptor antagonist.



In moderate grade OA synovium we did not always find a higher percentage of CD80 with respect to CD206. M1-like-activated macrophage factors IL1β, TNFα, IL6, MIP1α/CCL3, S100A8 and S100A9 were down-modulated both in contact and in transwell by ASC. However, in both systems ASC induced the typical M2-like macrophage markers IL10, CD163 and CD206. Activated-M1-like macrophages pre-treated with PGE2 receptor antagonist failed to decrease secretion of TNFα, IL6 and to increase that of IL10, CD163 and CD206 when co-cultured with ASC confirming a PGE2 specific role.



We demonstrated that ASC are responsible for the switching of activated-M1-like inflammatory macrophages to a M2-like phenotype, mainly through PGE2. This evidenced that activated-M1-like macrophages may represent a relevant cell model to test the efficacy/potency of ASC and suggests a specific role of ASC as important determinants in therapeutic dampening of synovial inflammation in OA.
SC Laboratorio di Immunoreumatologia e Rigenerazione Tissutale, Istituto Ortopedico Rizzoli, 40136 Bologna, Italy. Electronic address: cristina.manferdini@ior.it.
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Manferdini
SC Laboratorio di Immunoreumatologia e Rigenerazione Tissutale, Istituto Ortopedico Rizzoli, 40136 Bologna, Italy. Electronic address: francesca.paolella@ior.it.
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Paolella
SC Laboratorio di Immunoreumatologia e Rigenerazione Tissutale, Istituto Ortopedico Rizzoli, 40136 Bologna, Italy. Electronic address: elena.gabusi@ior.it.
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Gabusi
SD Laboratorio RAMSES, Istituto Ortopedico Rizzoli, 40136 Bologna, Italy. Electronic address: laura.gambari@ior.it.
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Gambari
SC Laboratorio di Immunoreumatologia e Rigenerazione Tissutale, Istituto Ortopedico Rizzoli, 40136 Bologna, Italy. Electronic address: anna.piacentini@ior.it.
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Piacentini
Laboratorio NABI, Istituto Ortopedico Rizzoli, 40136 Bologna, Italy. Electronic address: giuseppe.filardo@ior.it.
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Filardo
EFS-Pyrénéés-Méditerranéé, Toulouse F-31300, France. Electronic address: sandrine.fleury@efs.sante.fr.
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Fleury-Cappellesso
Department of Biomedicine, University Hospital Basel, University of Basel, Basel, Switzerland. Electronic address: abarbero@uhbs.ch.
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Barbero
Regenerative Medicine Institute, Galway, Ireland. Electronic address: mary.marphy@nuigalway.ie.
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Murphy
SC Laboratorio di Immunoreumatologia e Rigenerazione Tissutale, Istituto Ortopedico Rizzoli, 40136 Bologna, Italy. Electronic address: gina.lisignoli@ior.it.
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Lisignoli
D017667:Adipocytes / Q000187:drug effects*; D000328:Adult; D015703:Antigens, CD / Q000378:metabolism; D016022:Case-Control Studies; D002454:Cell Differentiation / Q000502:physiology; D002465:Cell Movement / Q000187:drug effects; D002478:Cells, Cultured; D015232:Dinoprostone / Q000494:pharmacology*; D005260:Female; D006801:Humans; D007963:Leukocytes, Mononuclear / Q000187:drug effects; D008264:Macrophages / Q000187:drug effects*; D008297:Male; D059630:Mesenchymal Stem Cells / Q000187:drug effects*; D008875:Middle Aged; D010003:Osteoarthritis / Q000473:pathology; D010120:Oxytocics / Q000494:pharmacology*; D050507:Subcutaneous Fat, Abdominal / Q000166:cytology; D013583:Synovial Membrane / Q000166:cytology / Q000187:drug effects
D015703:Antigens, CD; D010120:Oxytocics; D015232:Dinoprostone
Adipose mesenchymal stromal cells; Inflammation; Monocytes/macrophages; Osteoarthritis; Synovitis