title: Enhancement of proinflammatory and procoagulant responses to silica particles by monocyte-endothelial cell interactions.
abstract: BACKGROUND
  Inorganic particles, such as drug carriers or contrast agents, are often introduced into the vascular system. Many key components of the in vivo vascular environment include monocyte-endothelial cell interactions, which are important in the initiation of cardiovascular disease. To better understand the effect of particles on vascular function, the present study explored the direct biological effects of particles on human umbilical vein endothelial cells (HUVECs) and monocytes (THP-1 cells). In addition, the integrated effects and possible mechanism of particle-mediated monocyte-endothelial cell interactions were investigated using a coculture model of HUVECs and THP-1 cells. Fe₃O₄ and SiO₂ particles were chosen as the test materials in the present study.
  
  
  RESULTS
  The cell viability data from an MTS assay showed that exposure to Fe₃O₄ or SiO₂ particles at concentrations of 200 μg/mL and above significantly decreased the cell viability of HUVECs, but no significant loss in viability was observed in the THP-1 cells. TEM images indicated that with the accumulation of SiO₂ particles in the cells, the size, structure and morphology of the lysosomes significantly changed in HUVECs, whereas the lysosomes of THP-1 cells were not altered. Our results showed that reactive oxygen species (ROS) generation; the production of interleukin (IL)-6, IL-8, monocyte chemoattractant protein 1 (MCP-1), tumor necrosis factor (TNF)-α and IL-1β; and the expression of CD106, CD62E and tissue factor in HUVECs and monocytes were significantly enhanced to a greater degree in the SiO₂-particle-activated cocultures compared with the individual cell types alone. In contrast, exposure to Fe₃O₄ particles had no impact on the activation of monocytes or endothelial cells in monoculture or coculture. Moreover, using treatment with the supernatants of SiO₂-particle-stimulated monocytes or HUVECs, we found that the enhancement of proinflammatory response by SiO₂ particles was not mediated by soluble factors but was dependent on the direct contact between monocytes and HUVECs. Furthermore, flow cytometry analysis showed that SiO₂ particles could markedly increase CD40L expression in HUVECs. Our data also demonstrated that the stimulation of cocultures with SiO₂ particles strongly enhanced c-Jun NH2-terminal kinase (JNK) phosphorylation and NF-κB activation in both HUVECs and THP-1 cells, whereas the phosphorylation of p38 was not affected.
  
  
  CONCLUSIONS
  Our data demonstrate that SiO₂ particles can significantly augment proinflammatory and procoagulant responses through CD40-CD40L-mediated monocyte-endothelial cell interactions via the JNK/NF-κB pathway, which suggests that cooperative interactions between particles, endothelial cells, and monocytes may trigger or exacerbate cardiovascular dysfunction and disease, such as atherosclerosis and thrombosis. These findings also indicate that the monocyte-endothelial cocultures represent a sensitive in vitro model system to assess the potential toxicity of particles and provide useful information that may help guide the future design and use of inorganic particles in biomedical applications.
authors.affiliation: Shanghai Biomaterials Research & Testing Center, Shanghai Key Laboratory of Stomatology, Ninth People's Hospital, Shanghai Jiaotong University School of Medicine, No. 427 Ju-men Road, Shanghai 200023, China.
authors.forename: Xin
authors.initials: X
authors.lastname: Liu
authors.forename: Yang
authors.initials: Y
authors.lastname: Xue
authors.forename: Tingting
authors.initials: T
authors.lastname: Ding
authors.forename: Jiao
authors.initials: J
authors.lastname: Sun
mesh_terms: D015703:Antigens, CD / Q000378:metabolism; D015415:Biomarkers / Q000378:metabolism; D002448:Cell Adhesion / Q000187:drug effects / Q000502:physiology; D002450:Cell Communication / Q000187:drug effects / Q000502:physiology; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D018920:Coculture Techniques; D016207:Cytokines / Q000378:metabolism*; D005290:Ferric Compounds / Q000633:toxicity; D061307:Human Umbilical Vein Endothelial Cells / Q000187:drug effects* / Q000378:metabolism / Q000648:ultrastructure; D006801:Humans; D048031:JNK Mitogen-Activated Protein Kinases / Q000378:metabolism; D008247:Lysosomes / Q000187:drug effects / Q000648:ultrastructure; D008264:Macrophages / Q000187:drug effects* / Q000378:metabolism / Q000648:ultrastructure; D016328:NF-kappa B / Q000378:metabolism; D052638:Particulate Matter / Q000633:toxicity*; D015398:Signal Transduction; D012822:Silicon Dioxide / Q000633:toxicity*; D013925:Thromboplastin / Q000378:metabolism*
chemical_list: D015703:Antigens, CD; D015415:Biomarkers; D016207:Cytokines; D005290:Ferric Compounds; D016328:NF-kappa B; D052638:Particulate Matter; C000499:ferric oxide; D012822:Silicon Dioxide; D013925:Thromboplastin; D048031:JNK Mitogen-Activated Protein Kinases