Silver nanomaterials are increasingly being used as antimicrobial agents in medical devices. This study assessed the in vitro hemolytic potential of unbound silver particles in human blood to determine which physical and chemical particle properties contribute to mechanisms of red blood cell (RBC) damage. Four silver particle powders (two nano-sized and two micron-sized) were dispersed in water and characterized using transmission electron microscopy, dynamic light scattering, surface-enhanced Raman spectroscopy, and zeta potential measurement. Particle size and agglomeration were dependent on the suspension media. Under similar conditions to the hemolysis assay, with the particles added to phosphate buffered saline (PBS) and plasma, the size of the nanoparticles increased compared with particles suspended in water alone due to interaction with chloride ions and plasma proteins. To determine hemolysis response, aqueous particle suspensions were mixed with heparinized human blood diluted in PBS for 3.5 h at 37°C. Both nanoparticle preparations were significantly more hemolytic than micron-sized particles at equivalent mass concentrations > 220 μg/ml and at estimated surface area concentrations > 10 cm(2)/ml. The presence or absence of surface citrate on nanoparticles showed no significant difference in hemolysis. However, the aqueous nanoparticle preparations released significantly more silver ions than micron-sized particles, which correlated with increased hemolysis. Although significant size changes occurred to the silver particles due to interaction with media components, the higher level of in vitro hemolysis observed with nanoparticles compared with micron-sized particles may be related to their greater surface area, increased silver ion release, and direct interaction with RBCs.

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Photoluminescent silicon nanoparticles have a bright and stable fluorescence and are promising candidates for bio-imaging, cell staining and drug delivery. With increasing development of nanotechnology applications for biomedicine, an understanding of the potential toxicity of nanoparticles is needed to assess safety concerns for clinical applications. The objective of this study was to compare biological responses of silicon nanoparticles (SNs, 3 nm diameter) with silicon microparticles (SMs, approximately 100-3000 nm diameter) in cultured murine macrophages (RAW 264.7) using standard protocols for assessing cytotoxicity/cell viability and inflammatory responses developed for micron-sized particles. SNs and SMs were exposed to macrophages with and without addition of endotoxin lipopolysaccharide (LPS), a positive inducer of tumor necrosis factor-alpha (TNF-alpha), interleukin 6 (IL-6), and nitric oxide (NO). Cytotoxicity was assayed using the dye exclusion and MTT assays. Cell supernatants were assayed for production TNF-alpha, IL-6 and NO. SNs at concentrations < or = 20 microg ml(-1) exhibited no cytotoxicity or inflammatory responses; however, SNs and SMs >20 and 200 microg ml(-1), respectively, increased cytotoxicity compared with controls. SMs induced concentration-related increases in TNF-alpha and IL-6 production; in contrast, the production of these cytokines was shown to decrease with increasing concentrations of SNs. NO production was not induced by SNs or SMs alone. Fluorescence microscopy demonstrated that SNs were associated with the macrophages, either internalized or attached to cell membranes. In conclusion, evaluating differences in biological responses for nanoparticles compared with microparticles of the same material may help improve tests to assess biological responses of nanoparticles that may be used in biomedical applications.

More information characterizing the biological responses to nanoparticles is needed to allow the U.S. Food and Drug Administration to evaluate the safety and effectiveness of products with nano-scale components. The potential cytotoxicity and inflammatory responses of Au NPs (60 nm, NIST standard reference materials) were investigated in murine macrophages. Cytotoxicity was evaluated by MTT and LDH assays. Cytokines (IL-6, TNF-α), nitric oxide, and ROS were assayed to assess inflammatory responses. Morphological appearance and localization of particles were examined by high resolution illumination microscopy, transmission electron microscopy (TEM), and scanning TEM coupled with EDX spectroscopy. Results showed no cytotoxicity and no elevated production of proinflammatory mediators; however, imaging analyses demonstrated cellular uptake of Au NPs and localization within intracellular vacuoles. These results suggest that 60 nm Au NPs, under the exposure conditions tested, are not cytotoxic, nor elicit pro-inflammatory responses. The localization of Au NPs in intracellular vacuoles suggests endosomal containment and an uptake mechanism involving endocytosis.