Araujo, Nuna C. P.Viegas, CarlaZubía, EvaMagalhães, JoanaRamos, AcácioCarvalho, Maria M.Cruz, HenriqueSousa, João PauloBlanco, Francisco J.Vermeer, CeesSimes, Dina2021-01-142021-01-142020-12-07Marine Drugs 18 (12): 624 (2020)1660-3397http://hdl.handle.net/10400.1/14960Osteoarthritis (OA) remains a prevalent chronic disease without effective prevention and treatment. Amentadione (YP), a meroditerpenoid purified from the alga <i>Cystoseira usneoides</i>, has demonstrated anti-inflammatory activity. Here, we investigated the YP anti-osteoarthritic potential, by using a novel OA preclinical drug development pipeline designed to evaluate the anti-inflammatory and anti-mineralizing activities of potential OA-protective compounds. The workflow was based on in vitro primary cell cultures followed by human cartilage explants assays and a new OA co-culture model, combining cartilage explants with synoviocytes under interleukin-1β (IL-1β) or hydroxyapatite (HAP) stimulation. A combination of gene expression analysis and measurement of inflammatory mediators showed that the proposed model mimicked early disease stages, while YP counteracted inflammatory responses by downregulation of COX-2 and IL-6, improved cartilage homeostasis by downregulation of MMP3 and the chondrocytes hypertrophic differentiation factors Col10 and Runx2. Importantly, YP downregulated NF-κB gene expression and decreased phosphorylated IkBα/total IkBα ratio in chondrocytes. These results indicate the co-culture as a relevant pre-clinical OA model, and strongly suggest YP as a cartilage protective factor by inhibiting inflammatory, mineralizing, catabolic and differentiation processes during OA development, through inhibition of NF-κB signaling pathways, with high therapeutic potential.engOsteoarthritisAmentadionePreclinical osteoarthritis modelsMarine compoundsCystoseira usneoidesInflammationMineralizationChondrocytesSynoviocytesCartilage explantsjournal article2021-01-0810.3390/md18120624