C Manferdini1, F Paolella2, E Gabusi3, L Gambari4, A Piacentini5, G Filardo6, S Fleury-Cappellesso7, A Barbero8, M Murphy9, G Lisignoli10. 1. SC Laboratorio di Immunoreumatologia e Rigenerazione Tissutale, Istituto Ortopedico Rizzoli, 40136 Bologna, Italy. Electronic address: cristina.manferdini@ior.it. 2. SC Laboratorio di Immunoreumatologia e Rigenerazione Tissutale, Istituto Ortopedico Rizzoli, 40136 Bologna, Italy. Electronic address: francesca.paolella@ior.it. 3. SC Laboratorio di Immunoreumatologia e Rigenerazione Tissutale, Istituto Ortopedico Rizzoli, 40136 Bologna, Italy. Electronic address: elena.gabusi@ior.it. 4. SD Laboratorio RAMSES, Istituto Ortopedico Rizzoli, 40136 Bologna, Italy. Electronic address: laura.gambari@ior.it. 5. SC Laboratorio di Immunoreumatologia e Rigenerazione Tissutale, Istituto Ortopedico Rizzoli, 40136 Bologna, Italy. Electronic address: anna.piacentini@ior.it. 6. Laboratorio NABI, Istituto Ortopedico Rizzoli, 40136 Bologna, Italy. Electronic address: giuseppe.filardo@ior.it. 7. EFS-Pyrénéés-Méditerranéé, Toulouse F-31300, France. Electronic address: sandrine.fleury@efs.sante.fr. 8. Department of Biomedicine, University Hospital Basel, University of Basel, Basel, Switzerland. Electronic address: abarbero@uhbs.ch. 9. Regenerative Medicine Institute, Galway, Ireland. Electronic address: mary.marphy@nuigalway.ie. 10. SC Laboratorio di Immunoreumatologia e Rigenerazione Tissutale, Istituto Ortopedico Rizzoli, 40136 Bologna, Italy. Electronic address: gina.lisignoli@ior.it.
Abstract
OBJECTIVE: 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. DESIGN: 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. RESULTS: 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. CONCLUSIONS: 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.
OBJECTIVE: 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. DESIGN: 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. RESULTS: 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. CONCLUSIONS: 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.
Authors: Robert M Stefani; Saiti S Halder; Eben G Estell; Andy J Lee; Amy M Silverstein; Evie Sobczak; Nadeen O Chahine; Gerard A Ateshian; Roshan P Shah; Clark T Hung Journal: Tissue Eng Part A Date: 2018-10-31 Impact factor: 3.845
Authors: D Thompson; N Morrice; L Grant; S Le Sommer; K Ziegler; P Whitfield; N Mody; H M Wilson; M Delibegović Journal: Mol Metab Date: 2017-06-13 Impact factor: 7.422