title: Metalloproteinase and cytokine production by THP-1 macrophages following exposure to chitosan-DNA nanoparticles.
abstract: The use of nanoparticles for gene therapy is gaining more and more interest for medical applications. Chitosan is among the candidate polymers that have a potential application as a gene delivery system. Before using chitosan-DNA nanoparticles in vivo, one must study their interaction and cell's behavior. Since macrophages play an important role in inflammatory processes, this study was performed to investigate the effects of chitosan-DNA nanoparticles on human THP-1 cell line. Cytokine (TNF-alpha, IL-1beta, IL-6 and IL-10) and metalloproteinase (MMP-2 and MMP-9) release as well as their inhibitors (TIMP-1 and TIMP-2) were assessed after time course incubation with different amount of nanoparticles. Their secretion was quantified by enzyme-linked immunosorbent assay. Gelatinolytic activity of MMP-2 and MMP-9 was determined by zymography in cell supernatants and lysates. Cytokine secretion was not detected even in the presence of high amount of nanoparticles. On the contrary, the secretion of MMP-9 in cell supernatants increased significantly after 24 and 48 h in comparison with non-treated cells. MMP-2 secretion was augmented only after 48 h for the highest concentrations of nanoparticles (10 and 20 microg/ml DNA content). However, zymography studies showed that the secreted MMPs were in the proactive forms, while the active form of MMP-9, but not MMP-2, was detected in cell lysates when 10 and 20 microg/ml DNA containing nanoparticles were used. In conclusion, exposure of THP-1 macrophages to Ch-DNA nanoparticles did not induce release of proinflammatory cytokines. The presence of active MMP-9 within the macrophages could possibly be related to nanoparticle phagocytosis and degradation rather than to inflammatory reactions.
authors.affiliation: Biomedical Engineering Institute, Ecole Polytechnique de Montréal, C.P. 6079, Succursale Centre-ville, Montréal, Québec, Canada H3C 3A7. fatiha.chellat@polymtl.ca
authors.forename: Fatiha
authors.initials: F
authors.lastname: Chellat
authors.forename: Alexia
authors.initials: A
authors.lastname: Grandjean-Laquerriere
authors.forename: Richard
authors.initials: R
authors.lastname: Le Naour
authors.forename: Julio
authors.initials: J
authors.lastname: Fernandes
authors.forename: L'Hocine
authors.initials: L
authors.lastname: Yahia
authors.forename: Moncef
authors.initials: M
authors.lastname: Guenounou
authors.forename: Dominique
authors.initials: D
authors.lastname: Laurent-Maquin
mesh_terms: D002460:Cell Line; D048271:Chitosan / Q000737:chemistry / Q000276:immunology / Q000493:pharmacokinetics*; D016207:Cytokines / Q000276:immunology / Q000378:metabolism*; D004247:DNA / Q000737:chemistry / Q000276:immunology / Q000493:pharmacokinetics*; D005549:Foreign-Body Reaction / Q000276:immunology / Q000378:metabolism* / Q000473:pathology; D006801:Humans; D008264:Macrophages / Q000276:immunology / Q000378:metabolism*; D020782:Matrix Metalloproteinases / Q000276:immunology / Q000378:metabolism*; D043942:Nanotubes / Q000737:chemistry / Q000648:ultrastructure; D010316:Particle Size
chemical_list: D016207:Cytokines; D004247:DNA; D048271:Chitosan; D020782:Matrix Metalloproteinases