Extracellular Matrix Protection Factor‐1, a Novel Osteoarthritis Therapeutic, Decreases Expression of Interleukin‐1 beta by Primary Cultures of Human Osteoarthritic Chondrocytes

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Osteoarthritis (OA) is characterized by an imbalance of cartilage extracellular matrix (ECM) production and degradation, due, in part, to increased cytokine production. It has been demonstrated that the cytokine, Interleukin‐1 beta (IL‐1β), is produced by OA articular chondrocytes and plays a prominent role in altering ECM metabolism by targeting the matrix metalloproteases (MMPs) that degrade the ECM. We are developing a novel, OA therapeutic, Extracellular Matrix Protection Factor‐1 (ECPF‐1) that targets MMP interaction with its ECM substrate. We have demonstrated a reduction in ECM degradation when primary, human OA chondrocyte (HOAC) cultures are treated with ECPF‐1. In this study, HOACs isolated from total knee arthroplasty were reared in three‐dimensional, serum‐free, alginate cultures. The sides of least and greatest pathology of the femoral condyles and tibial plateaus were determined by gross inspection and cells isolated were cultured separately (LP and GP). Chondrocytes were plated at 2.5×106 cells per milliliter of alginate and incubated in serum‐free medium for five days. Cells were released from the alginate and the RNA extracted from the pellet, reverse‐transcribed into cDNA and quantitative, real time PCR (qRT‐PCR) was performed with TaqMan Gene Expression assays for IL‐1β, collagen type I, collagen type II and 18s rRNA. In a previous study, IL‐1β production was detected in the conditioned media of HOAC cultures with the highest concentration of this cytokine present in the cultures established from the region of greatest pathology (GP). In this study, IL‐1β expression was detectable in mRNA produced by LP and GP HOAC cultures. The expression was reduced 2‐fold in LP cultures treated 24 hours with 2.5 uM ECPF‐1 and 1.4 fold in GP cultures. These results demonstrate the ability of ECPF‐1 to alter expression of a molecule involved in the feedback loop of ECM destruction. ECPF‐1 provides a tool with which to further define the cellular mechanisms responsible for OA chondrocyte pathology.Support or Funding InformationNew Jersey Health Foundation, Center for Chronic Disorders of Aging Small Grant, PCOM's Division of ResearchThis abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.