Viegas, CarlaAraújo, NunaCarreira, JoanaPontes, Jorge FilipeMacedo, Anjos L.Vinhas, MauríciaMoreira, Ana S.Faria, Tiago Q.Grenha, Anade Matos, António A.Schurgers, LeonVermeer, CeesSimes, Dina2022-06-282022-06-282022-04-27International Journal of Molecular Sciences 23 (9): 4813 (2022)1422-0067http://hdl.handle.net/10400.1/17888Chronic inflammation is a major driver of chronic inflammatory diseases (CIDs), with a tremendous impact worldwide. Besides its function as a pathological calcification inhibitor, vitamin K-dependent protein Gla-rich protein (GRP) was shown to act as an anti-inflammatory agent independently of its gamma-carboxylation status. Although GRP’s therapeutic potential has been highlighted, its low solubility at physiological pH still constitutes a major challenge for its biomedical application. In this work, we produced fluorescein-labeled chitosan-tripolyphosphate nanoparticles containing non-carboxylated GRP (ucGRP) (FCNG) via ionotropic gelation, increasing its bioavailability, stability, and anti-inflammatory potential. The results indicate the nanosized nature of FCNG with PDI and a zeta potential suitable for biomedical applications. FCNG’s anti-inflammatory activity was studied in macrophage-differentiated THP1 cells, and in primary vascular smooth muscle cells and chondrocytes, inflamed with LPS, TNFα and IL-1β, respectively. In all these in vitro human cell systems, FCNG treatments resulted in increased intra and extracellular GRP levels, and decreased pro-inflammatory responses of target cells, by decreasing pro-inflammatory cytokines and inflammation mediators. These results suggest the retained anti-inflammatory bioactivity of ucGRP in FCNG, strengthening the potential use of ucGRP as an anti-inflammatory agent with a wide spectrum of application, and opening up perspectives for its therapeutic application in CIDs.engNanoparticlesGla-rich protein (GRP)Chronic inflammatory diseases (CIDs)InflammationVitamin K-dependent protein (VKDP)journal article2022-05-1210.3390/ijms23094813