title: iNOS-mediated generation of reactive oxygen and nitrogen species by biomaterial-adherent neutrophils.
abstract: Infection due to implanted cardiovascular biomaterials is a serious complication initiated by bacterial adhesion to the surface of the implant. The release of reactive oxygen species by neutrophils, particularly superoxide anion, is a well-known bactericidal mechanism. Additionally, nitric oxide (NO) has also been identified as an important cytotoxic mediator in acute and chronic inflammatory responses with enhanced NO production by upregulation of inducible nitric oxide synthase (iNOS). The interaction of NO and superoxide anion will result in the formation of peroxynitrite (OONO-), a potent cytotoxic oxidant. In this study, we have shown that biomaterial-induced neutrophil activation does not cause upregulation of iNOS and activation of iNOS-mediated pathways. However, NO and O2- production does occur over time upon adhesion to a biomaterial and is modulated by biomaterial surface chemistry. With no stimulus, the polyethylene oxide-modified polyurethane induced greater neutrophil activation than did the control as indicated by the increased production of NO and O2- over time. Adherent-stimulated neutrophils generally produced lower amounts of NO over time in comparison with unstimulated cells. Furthermore, there is no evidence of peroxynitrite activity in unstimulated neutrophils adherent to the Elasthane 80A. However, upon stimulation with adherent Staphylococcus epidermidis, peroxynitrite formation did occur. Our results suggest that bactericidal mechanisms in neutrophils involving NO generation (NOS pathway) are further compromised than O2- producing pathways (NADPH oxidase) upon exposure to biomaterials, resulting in a diminished microbial killing capacity, which can increase the probability of device-centered infections.
authors.affiliation: Department of Biomedical Engineering, Case Western Reserve University, Cleveland, Ohio 44106, USA. jma6@po.cwru.edu
authors.forename: Jasmine D
authors.initials: JD
authors.lastname: Patel
authors.forename: Tianyi
authors.initials: T
authors.lastname: Krupka
authors.forename: James M
authors.initials: JM
authors.lastname: Anderson
mesh_terms: D000900:Anti-Bacterial Agents; D001672:Biocompatible Materials / Q000009:adverse effects* / Q000737:chemistry*; D002448:Cell Adhesion; D006801:Humans; D018375:Neutrophil Activation; D009504:Neutrophils / Q000166:cytology* / Q000201:enzymology / Q000378:metabolism*; D052247:Nitric Oxide Synthase Type II / Q000378:metabolism*; D030421:Peroxynitrous Acid / Q000032:analysis / Q000378:metabolism; D011092:Polyethylene Glycols / Q000494:pharmacology; D011140:Polyurethanes / Q000494:pharmacology; D026361:Reactive Nitrogen Species / Q000378:metabolism*; D017382:Reactive Oxygen Species / Q000378:metabolism*; D013210:Staphylococcus / Q000187:drug effects
chemical_list: D000900:Anti-Bacterial Agents; D001672:Biocompatible Materials; D011140:Polyurethanes; D026361:Reactive Nitrogen Species; D017382:Reactive Oxygen Species; D030421:Peroxynitrous Acid; D011092:Polyethylene Glycols; D052247:Nitric Oxide Synthase Type II