Revealing Silver Nanoparticle Uptake by Macrophages Using SR-µXRF and LA-ICP-MS.

To better study the impact of nanoparticles on both in vitro and in vivo models, tissue distribution and cellular doses need to be described more closely. Here silver nanoparticles were visualized in alveolar macrophages by means of synchrotron radiation micro X-ray fluorescence spectroscopy (SR-µXRF) with high spatial resolution of 3 × 3 µm2. For the spatial allocation of silver signals to cells and tissue structures, additional elemental labeling was carried out by staining with eosin, which binds to protein and can be detected as bromine signal with SR-µXRF. The method was compatible with immunostaining of macrophage antigens. We found that the silver distribution obtained with SR-µXRF was largely congruent with distribution maps from a subsequent laser ablation inductively coupled plasma mass spectrometry (LA-ICP-MS) of the same tissue sites. The study shows a predominant, though not exclusive uptake of silver into alveolar macrophages in the rat lung, which can be modeled by a similar uptake in cultured alveolar macrophages. Advantages and limitations of the different strategies for measuring nanoparticle uptake at the single cell level are discussed.

Phosphonate coating of SiO2 nanoparticles abrogates inflammatory effects and local changes of the lipid composition in the rat lung: a complementary bioimaging study.

BACKGROUND: The well-known inflammatory and fibrogenic changes of the lung upon crystalline silica are accompanied by early changes of the phospholipid composition (PLC) as detected in broncho-alveolar lavage fluid (BALF). Amorphous silica nanoparticles (NPs) evoke transient lung inflammation, but their effect on PLC is unknown. Here, we compared effects of unmodified and phosphonated amorphous silica NP and describe, for the first time, local changes of the PLC with innovative bioimaging tools. METHODS: Unmodified (SiO2-n), 3-(trihydroxysilyl) propyl methylphosphonate coated SiO2-n (SiO2-p) as well as a fluorescent surrogate of SiO2-n (SiO2-FITC) nanoparticles were used in this study. In vitro toxicity was tested with NR8383 alveolar macrophages. Rats were intratracheally instilled with SiO2-n, SiO2-p, or SiO2-FITC, and effects on lungs were analyzed after 3 days. BALF from the right lung was analyzed for inflammatory markers. Cryo-sections of the left lung were subjected to fluorescence microscopy and PLC analyses by matrix-assisted laser desorption/ionization mass spectrometry imaging (MALDI-MS), Fourier transform infrared microspectroscopy (FT-IR), and tandem mass spectrometry (MS/MS) experiments. RESULTS: Compared to SiO2-p, SiO2-n NPs were more cytotoxic to macrophages in vitro and more inflammatory in the rat lung, as reflected by increased concentration of neutrophils and protein in BALF. Fluorescence microscopy revealed a typical patchy distribution of SiO2-FITC located within the lung parenchyma and alveolar macrophages. Superimposable to this particle distribution, SiO2-FITC elicited local increases of phosphatidylglycerol (PG) and phosphatidylinositol (PI), whereas phoshatidylserine (PS) and signals from triacylgyceride (TAG) were decreased in the same areas. No such changes were found in lungs treated with SiO2-p or particle-free instillation fluid. CONCLUSIONS: Phosphonate coating mitigates effects of silica NP in the lung and abolishes their locally induced changes in PLC pattern. Bioimaging methods based on MALDI-MS may become a useful tool to investigate the mode of action of NPs in tissues.

Impact of particle size and surface modification on gold nanoparticle penetration into human placental microtissues.

AIM: Nanoparticle-based drug carriers hold great promise for the development of targeted therapies in pregnancy with reduced off-target effects. Here, we performed a mechanistic in vitro study on placental localization and penetration of gold nanoparticles (AuNPs) in dependence of particle size and surface modification. MATERIALS & METHODS: AuNP uptake and penetration in human placental coculture microtissues was assessed by inductively coupled plasma-mass spectrometry, transmission electron microscopy and laser ablation-inductively coupled plasma-mass spectrometry. RESULTS: Higher uptake and deeper penetration was observed for smaller (3-4 nm) or sodium carboxylate-modified AuNPs than for larger (13-14 nm) or PEGylate AuNPs, which barely passed the trophoblast barrier layer. CONCLUSION: It is possible to steer placental uptake and penetration of AuNPs by tailoring their properties, which is a prerequisite for the development of targeted therapies in pregnancy.

Dielectric analysis of depth dependent curing behavior of dental resin composites.

OBJECTIVES: The aim of this study is to investigate depth dependent changes of polymerization process and kinetics of visible light-curing (VLC) dental composites in real-time. The measured quantity - "ion viscosity" determined by dielectric analysis (DEA) - provides the depth dependent reaction rate which is correlated to the light intensity available in the corresponding depths derived from light transmission measurements. METHODS: The ion viscosity curves of two composites (VOCO Arabesk Top and Grandio) were determined during irradiation of 40s with a light-curing unit (LCU) in specimen depths of 0.5/0.75/1.0/1.25/1.5/1.75 and 2.0mm using a dielectric cure analyzer (NETZSCH DEA 231 with Mini IDEX sensors). The thickness dependent light transmission was measured by irradiation composite specimens of various thicknesses on top of a radiometer setup. RESULTS: The shape of the ion viscosity curves depends strongly on the specimen thickness above the sensor. All curves exhibit a range of linear time dependency of the ion viscosity after a certain initiation time. The determined initiation times, the slopes of the linear part of the curves, and the ion viscosities at the end of the irradiation differ significantly with depth within the specimen. The slopes of the ion viscosity curves as well as the light intensity values decrease with depth and fit to the Lambert-Beer law. The corresponding attenuation coefficients are determined for Arabesk Top OA2 to 1.39mm(-1) and 1.48mm(-1), respectively, and for Grandio OA2 with 1.17 and 1.39mm(-1), respectively. For thicknesses exceeding 1.5mm a change in polymerization behavior is observed as the ion viscosity increases subsequent to the linear range indicating some kind of reaction acceleration. SIGNIFICANCE: The two VLC composites and different specimen thicknesses discriminate significantly in their ion viscosity evolution allowing for a precise characterization of the curing process even with respect to the polymerization mechanism.

Curing kinetics of visible light curing dental resin composites investigated by dielectric analysis (DEA).

During the curing process of light curing dental composites the mobility of molecules and molecule segments is reduced leading to a significant increase of the viscosity as well as the ion viscosity. Thus, the kinetics of the curing behavior of 6 different composites was derived from dielectric analysis (DEA) using especially redesigned flat sensors with interdigit comb electrodes allowing for irradiation at the top side and measuring the ion viscosity at the bottom side. As the ion viscosities of dental composites change 1-3 orders of magnitude during the curing process, DEA provides a sensitive approach to evaluate their curing behavior, especially in the phase of undisturbed chain growth. In order to determine quantitative kinetic parameters a kinetic model is presented and examined for the evaluation of the ion viscosity curves. From the obtained results it is seen that DEA might be employed in the investigation of the primary curing process, the quality assurance of ingredients as well as the control of processing stability of the light curing dental composites.