Conditioned media from human osteoarthritic synovium induces inflammation in a synoviocyte cell line

Elisa Belluzzi, Eleonora Olivotto, Giovanna Toso, Augusto Cigolotti, Assunta Pozzuoli, Carlo Biz, Giovanni Trisolino, Pietro Ruggieri, Brunella Grigolo, Roberta Ramonda, Marta Favero

Research output: Contribution to journalArticlepeer-review

Abstract

AIM: Osteoarthritis (OA) is a whole joint pathology involving cartilage, synovial membrane, meniscus, subchondral bone and infrapatellar fat pad (IFP). Synovitis has been widely documented in OA suggesting its important role in pathogenesis. The aim of this study was to investigate the role of different joint tissues in promoting synovitis.

MATERIALS AND METHODS: Conditioned media (CM) from cartilage, synovial membrane, meniscus and IFP, were generated from tissues of five patients undergoing total knee replacement and used to stimulate a human fibroblast-like synoviocytes cell line (K4IM). Cytokines, chemokines and metalloproteases release was analyzed in all CM by Bio-Plex Assay and sulfated glycosaminoglycan (GAG) content by dimethylmethylene blue assay. Gene expression of several markers was evaluated by Real-Time PCR in K4IM cells stimulated with the CM obtained from joint tissues.

RESULTS: CM from all tissues produced high levels of IL-6, IL-8, and CCL2. CCL21, MMP-3 and -13 levels were detected in all CM except IFP. MMP-10 was present only in CM of cartilage and synovial tissues. IL-1β, IL-15, TNF-α, CCL5 and CCL19 were undetectable. However, only K4IM cells stimulated by the CM from OA synovium showed an increase of IL-6, CXCL-8, CCL21, MMP10 and IL-1β expression.

CONCLUSIONS: Our study showed that K4IM might be a suitable in vitro model for evaluating different cellular pathways in OA studies. Importantly, we demonstrated that in OA, all joint tissues might be involved in the progression of synovitis with a predominant role of synovial membrane itself compared to the other joint tissues.

Original languageEnglish
Pages (from-to)1-10
Number of pages10
JournalConnective Tissue Research
DOIs
Publication statusE-pub ahead of print - May 2018

Keywords

  • Conditioned media
  • infrapatellar fat pad
  • cartilage
  • inflammation
  • meniscus
  • osteoarthritis

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