ABSTRACT
Enzyme-linked immunosorbent assay (ELISA) for monitoring the effects of nanoparticles on immune cells is a conventional method of assessing the levels of cytokine that are released into the culture supernatant after the addition of nanoparticles to a macrophage culture. However, it has been found that the presence of nanoparticles can interfere with spectrophotometric analysis, used as an indicator test system; thus, it is necessary to thoroughly checked for the possibility of interference. In this chapter, the assessment method of cytokine production is covered in detail by utilizing the cytokine model produced by silver nanoparticles.
Subject(s)
Enzyme-Linked Immunosorbent Assay/methods , Interleukin-8/biosynthesis , Metal Nanoparticles/toxicity , Silver/toxicity , Humans , Reference Standards , Specimen Handling , Spectrophotometry , U937 CellsABSTRACT
Because of the limited information on size-dependent particle-mediated effects, the present study was conducted to determine if the changes in induced protein expression between 5 nm silver nanoparticles and 100 nm particles after exposure to sub-lethal concentrations. A total of 28,000 cDNA profiles were screened using 5 nm silver nanoparticles and 100 nm silver nanoparticles in a macrophage cell line. Based on results obtained from cDNA microarray we also assessed protein levels of hemeoxygenase-1 (HO-1), heat shock protein-70 (HSP-70) and interleukin-8 (IL-8), which were shown to significantly increase. Together with results obtained using N-acetylcystein (NAC), we were able to clearly show that low level and early stage exposure to 5 nm silver nanoparticles, but not 100 nm, induces expression of IL-8 as well as stress genes against reactive oxygen species (ROS). Therefore, we provide important data to understand and identify the early effects of silver nanoparticles on the immune system.
Subject(s)
Macrophages/drug effects , Macrophages/metabolism , Metal Nanoparticles/therapeutic use , Oligonucleotide Array Sequence Analysis/methods , Silver/therapeutic use , Cell Line , Cell Survival/drug effects , HSP70 Heat-Shock Proteins/metabolism , Heme Oxygenase-1/metabolism , Humans , Interleukin-8/metabolism , Metal Nanoparticles/adverse effects , Metal Nanoparticles/chemistry , Metal Nanoparticles/ultrastructure , Microscopy, Electron, Transmission , Reactive Oxygen Species/metabolism , Real-Time Polymerase Chain Reaction , Silver/adverse effects , Silver/chemistryABSTRACT
Silver nanoparticles (AgNPs) are one of the most commonly used nanomaterials due to their antibacterial properties. In this study, we examined the effects of polyvinylpyrrolidone (PVP)-coated AgNPs (average size 2.3nm) on angiogenesis in both an in vivo model and an in vitro endothelial cell line, SVEC4-10. Increased angiogenesis was detected around the injection site of AgNP-containing Matrigel in vivo. AgNPs also increased the infiltration of endothelial cells and the hemoglobin (Hb) content in AgNP-Matrigel plugs implanted into mice. AgNPs induced endothelial cell tube formation on growth factor-reduced Matrigel, production of reactive oxygen species (ROS), and production of angiogenic factors, such as vascular endothelial growth factor (VEGF) and nitric oxide (NO), in SVEC4-10 cells. In addition, AgNPs promoted the activation of FAK, Akt, ERK1/2, and p38, which are all involved in VEGF receptor (VEGFR)-mediated signaling. Finally, AgNP-treated tumors caused angiogenesis around tumors in B16F10 melanomas after they were injected into mice, and the Hb concentration in the tumors increased in a concentration-dependent manner with AgNP treatment. Thus, our study suggests that exposure to AgNPs can cause angiogenesis through the production of angiogenic factors.
Subject(s)
Endothelium, Vascular/drug effects , Metal Nanoparticles/toxicity , Neovascularization, Pathologic/chemically induced , Povidone/toxicity , Silver/toxicity , Animals , Cell Line , Drug Implants , Endothelium, Vascular/metabolism , Female , Hemoglobins/metabolism , Melanoma, Experimental/blood supply , Melanoma, Experimental/metabolism , Metal Nanoparticles/administration & dosage , Metal Nanoparticles/chemistry , Mice , Mice, Inbred C57BL , Neovascularization, Pathologic/metabolism , Nitric Oxide/metabolism , Particle Size , Povidone/administration & dosage , Povidone/chemistry , Reactive Oxygen Species/metabolism , Receptors, Vascular Endothelial Growth Factor/metabolism , Signal Transduction/drug effects , Silver/administration & dosage , Silver/chemistry , Subcutaneous Tissue/blood supply , Subcutaneous Tissue/drug effects , Subcutaneous Tissue/metabolism , Subcutaneous Tissue/pathology , Surface Properties , Vascular Endothelial Growth Factors/metabolismABSTRACT
The immune-response of macrophages is an important area of investigation since it represents the major pathway by which early-stage defense barriers are established in skin, lungs, and mucosal systems to counteract foreign objects. In this study, we have examined the size-dependent inflammatory and toxicological effects of nanostructured silver particles (nano-Ag) on macrophage immune cells.
Subject(s)
Macrophages/drug effects , Metal Nanoparticles/chemistry , Metal Nanoparticles/toxicity , Particle Size , Silver/chemistry , Silver/toxicity , Cell Line , Humans , Inflammation/chemically induced , Inflammation/metabolism , Interleukin-8/biosynthesis , Macrophages/immunology , Macrophages/metabolism , Phagocytosis/drug effects , Phagocytosis/immunology , Reactive Oxygen Species/metabolismABSTRACT
As a nanotechnology has been actively applied to the overall areas of scientific fields, it is necessary to understand the characteristic features, physical behaviors and the potential effects of exposure to nanomaterials and their toxicity. In this article we review the immunological influences induced by several nanomaterials and emphasize establishment of the animal models to estimate the impact of these nanomaterials on development of immunological diseases.
