These particles BGJ398 ic50 have the additional advantage that they also allow the determination of the deposition rate in their core-fluorescently labeled form. Multi-walled carbon nanotubes with different diameters were used to identify the influence of shape and the aggregation behavior of the nanoparticles since carbon nanotubes show high aggregation and polystyrene particles low aggregation (Wiesner and Colvin, 2005). Carbon nanotubes are also candidates for numerous medical applications (Zhang et al., 2010). 20, 40, 100, and 200 nm red (580/605) fluorescent labeled carboxyl-functionalized polystyrene particles (FluoSpheres)
were purchased from Invitrogen (Vienna). Carboxylated short multi-walled carbon nanotubes (0.5–2 μm long, purity >95%) with outer diameters <8, 20–30 and >50 nm were obtained from CheapTubes Inc. (Brattleboro, Vermont). To identify a potential difference in cytotoxicity between exposure in submersed culture and exposure as aerosol, 20 nm amine-functionalized polystyrene particles were used (Estapor Microspheres, Merck Chimie S.A.S., Fontenay-sous-Bois). For exposure all nanoparticles were diluted and the suspensions were put into
an Elmasonic http://www.selleckchem.com/products/sch772984.html S40 water bath (ultrasonic frequency: 37 kHz, Elma, Singen) for 20 min prior to all experiments. For the VITROCELL system and the MicroSprayer cells were cultured for 24 h after the exposures. Fluorescein sodium salt (Sigma Aldrich, Steinheim) was used as a macromolecular reference substance. Nanoparticle-sizes were determined routinely tuclazepam by dynamic light scattering using a Zetasizer Nano ZS (Malvern Instruments, Malvern) equipped
with a 532 nm HeNe laser, taking into account viscosity as well as refraction index. Polystyrene particles were diluted with the same solvent used for the exposures (distilled water for VITROCELL PT/PARI BOY LC Sprint system and DMEM + 10% FBS for MicroSprayer) to a concentration of 200 μg/ml and sonicated for 20 min before measurements. To study the stability of the nanoparticles in the VITROCELL PT/PARI BOY LC Sprint system samples of the condensate from the vial at the end of the glass tube (Fig. 1a) were also tested. After equilibration of the sample solution to 25 °C, scattered light was detected at a 173° angle with laser attenuation and the dynamic fluctuations of light scattering intensity caused by Brownian motion of the particles was evaluated. Polydispersity Indices <0.2 are interpreted as indication for homogenous samples (Stancampiano et al., 2008). The viscosities of the different solutions were 0.88 cP (water) and 0.94 cP (DMEM + 10% FBS). Additionally, the refraction indices of all investigated media were found to be around 1.33 (1.33 for water, 1.345 DMEM + 10% FBS. These values were very similar and showed no impact on the measured particle sizes. CNTs were suspended in DMEM + 10% FBS at 1 mg/ml.