Ta-Chih Hsiao

Effects of size and surface of zinc oxide and aluminum-doped zinc oxide nanoparticles on cell viability inferred by proteomic analyses

Although the health effects of zinc oxide nanoparticles (ZnONPs) on the respiratory system have been reported, the fate, potential toxicity, and mechanisms in biological cells of these particles, as related to particle size and surface characteristics, have not been well elucidated. To determine the physicochemical properties of ZnONPs that govern cytotoxicity, we investigated the effects of size, electronic properties, zinc concentration, and pH on cell viability using human alveolar-basal epithelial A549 cells as a model. We observed that a 2-hour or longer exposure to ZnONPs induced changes in cell viability. The alteration in cell viability was associated with the zeta potentials and pH values of the ZnONPs. Proteomic profiling of A549 exposed to ZnONPs for 2 and 4 hours was used to determine the biological mechanisms of ZnONP toxicity. p53-pathway activation was the core mechanism regulating cell viability in response to particle size. Activation of the Wnt and TGFβ signaling pathways was also important in the cellular response to ZnONPs of different sizes. The cadherin and Wnt signaling pathways were important cellular mechanisms triggered by surface differences. These results suggested that the size and surface characteristics of ZnONPs might play an important role in their observed cytotoxicity. This approach facilitates the design of more comprehensive systems for the evaluation of nanoparticles.

Allergenicity and toxicology of inhaled silver nanoparticles in allergen-provocation mice models

Silver nanoparticles (AgNP) have been associated with the exacerbation of airway hyperresponsiveness. However, the allergenicity and toxicology of AgNP in healthy and allergic individuals are unclear. We investigated the pathophysiological responses to AgNP inhalation in a murine model of asthma. Continuous and stable levels of 33 nm AgNP were maintained at 3.3 mg/m3 during the experimental period. AgNP exposure concomitant with ovalbumin challenge increased the enhanced pause (Penh) in the control and allergic groups. AgNP evoked neutrophil, lymphocyte and eosinophil infiltration into the airways and elevated the levels of allergic markers (immunoglobulin E [IgE] and leukotriene E4 [LTE4]), the type 2 T helper (Th2) cytokine interleukin-13 (IL-13), and oxidative stress (8-hydroxy-2′-deoxyguanosine [8-OHdG]) in healthy and allergic mice. Bronchocentric interstitial inflammation was observed after AgNP inhalation. After inhalation, the AgNP accumulated predominantly in the lungs, and trivial amounts of AgNP were excreted in the urine and feces. Furthermore, the AgNP induced inflammatory responses in the peritoneum. The inhalation of AgNP may present safety concerns in healthy and susceptible individuals.

Aqueous film formation on irregularly shaped inorganic nanoparticles before deliquescence, as revealed by a hygroscopic differential mobility analyzer–Aerosol particle mass system

ABSTRACT A hygroscopic tandem differential mobility analyzer (H-TDMA) and a hygroscopic coupled DMA and aerosol particle mass (H-DMA-APM) were coupled to examine aqueous film formation and the deliquescence behavior of inorganic nanoparticles. The two systems complement each other because H-DMA-APM measures mass change, while H-TDMA measures mobility diameter (volume) change of nanoparticles upon water uptake. The former mass change was, in particular, more capable to discern minute particle phase changes than the latter size change at moderate RHs. The mass and diameter changes were used to derive the particle effective density for evaluation of aqueous film formation on the nanoparticle surface before and after deliquescence transition. The measurements further showed that approximately 3–5 and 12–20 monolayer equivalents of water molecules formed on the respective surface of 50- and 100-nm inorganic aerosols (ammonium sulfate and sodium chloride) before deliquescence relative humidity (DRH). These findings support the physical basis of the coated-surface model given by Russell and Ming in 2002, and suggest that the phase transition of inorganic nanoparticles near deliquescence is a gradual process instead of an abrupt change. This phenomenon changed the surface energy values, thus confirming the explanation that the DRH of nanoparticles increases as the particle size decreases. This is the first direct observation of nanoparticle deliquescence phase transition using the H-DMA-APM system, and the detailed characterization of aqueous film formation on inorganic nanoparticles is feasible with the presented measurement systems.

Filter Quality of Pleated Filter Cartridges

The performance of dust cartridge filters commonly used in dust masks and in room ventilation depends both on the collection efficiency of the filter material and the pressure drop across the filter. Currently, the optimization of filter design is based only on minimizing the pressure drop at a set velocity chosen by the manufacturer. The collection efficiency, an equally important factor, is rarely considered in the optimization process. In this work, a filter quality factor, which combines the collection efficiency and the pressure drop, is used as the optimization criterion for filter evaluation. Most respirator manufacturers pleat the filter to various extents to increase the filtration area in the limit space within the dust cartridge. Six sizes of filter holders were fabricated to hold just one pleat of filter, simulating six different pleat counts, ranging from 0.5 to 3.33 pleats cm(-1). The possible electrostatic charges on the filter were removed by dipping in isopropyl alcohol, and the air velocity is fixed at 100 cm s(-1). Liquid dicotylphthalate particles generated by a constant output atomizer were used as challenge aerosols to minimize particle loading effects. A scanning mobility particle sizer was used to measure the challenge aerosol number concentrations and size distributions upstream and downstream of the pleated filter. The pressure drop across the filter was monitored by using a calibrated pressure transducer. The results showed that the performance of pleated filters depend not only on the size of the particle but also on the pleat count of the pleated filter. Based on filter quality factor, the optimal pleat count (OPC) is always higher than that based on pressure drop by about 0.3-0.5 pleats cm(-1). For example, the OPC is 2.15 pleats cm(-1) from the standpoint of pressure drop, but for the highest filter quality factor, the pleated filter needed to have a pleat count of 2.65 pleats cm(-1) at particle diameter of 122 nm. From the aspect of filter quality factor, this study suggests that the respirator manufacturers should add approximately 0.5 pleats cm(-1) to the OPC derived from the generalized correlation curve for pleated filter design based on minimum pressure drop.

Surface PEGylation of Silver Nanoparticles: Kinetics of Simultaneous Surface Dissolution and Molecular Desorption

A quantitative study of the stability of silver nanoparticles (AgNPs) conjugated with thiolated polyethylene glycol (SH-PEG) was conducted using gas-phase ion-mobility and mass analyses. The extents of aggregation and surface dissolution of AgNPs, as well as the amount of SH-PEG adsorption and desorption, were able to be characterized simultaneously for the kinetic study. The results show that the SH-PEG with a molecular mass of 6 kg/mol (SH-PEG6K) was able to adsorb to the surface of AgNP to form PEG6K-HS-AgNP conjugates, with the maximum surface adsorbate density of ∼0.10 nm(-2). The equilibrium binding constant for SH-PEG6K on AgNPs was calculated as ∼(4.4 ± 0.9) × 10(5) L/mol, suggesting a strong affinity due to thiol bonding to the AgNP surface. The formation of SH-PEG6K corona prevented PEG6K-HS-AgNP conjugates from aggregation under the acidic environment (pH 1.5), but dissolution of core AgNPs occurred following a first-order reaction. The rate constant of Ag dissolution from PEG6K-HS-AgNP was independent of the starting surface packing density of SH-PEG6K on AgNP (σ0), indicating that the interactions of H(+) with core AgNP were not interfered by the presence of SH-PEG6K corona. The surface packing density of SH-PEG6K decreased simultaneously following a first-order reaction, and the desorption rate constant of SH-PEG6K from the conjugates was proportional to σ0. Our work presents the first quantitative study to illustrate the complex mechanism that involves simultaneous aggregation and dissolution of core AgNPs in combination with adsorption and desorption of SH-PEG. This work also provides a prototype method of coupled experimental scheme to quantify the change of particle mass versus the corresponding surface density of functional molecular species on nanoparticles.

Pulmonary pathobiology induced by zinc oxide nanoparticles in mice: A 24-hour and 28-day follow-up study

ABSTRACT Inhaled zinc oxide nanoparticles (ZnONPs) have high deposition rates in the alveolar region of the lungs; however, the adverse health effects of ZnONPs on the respiratory system are unclear. Herein, pathobiological responses of the respiratory system of mice that received intratracheal administration of ZnONPs were investigated by a combination of molecular and imaging (SPECT and CT) approaches. Also, normal BEAS‐2B and adenocarcinoma A549 cells were used to confirm the results in mice. First, female BALB/c mice were administrated a series of doses of 20‐nm ZnONPs and were compared to the phosphate‐buffered saline control for 24‐h and 28‐day follow‐up observations. Field emission‐scanning electron microscopy and an energy‐dispersive X‐ray microanalysis were first used to characterize ZnONPs. After 24 h, instilled ZnONPs had caused significant increases in lactic dehydrogenase (LDH) in bronchoalveolar lavage fluid (BALF) and 8‐hydroxy‐2′‐deoxyguanosine (8‐OHdG), caspase‐3, and the p63 tumor marker in lung tissues (p < 0.05). Airway inflammation was present in a dose‐dependent manner from the upper to the lower airway as analyzed by SPECT. After 28 days, p63 had significantly increased due to ZnONP exposure in lung tissues (p < 0.05). Pulmonary inflammatory infiltration mainly occurred in the left and right subsegments of the secondary bronchial bifurcation as observed by CT. A significant increase in p63 and decrease in TTF1 levels were observed in BEAS‐2B cells by ZnONP (p < 0.05), but not in A549 cells. Our results demonstrated that regional lung inflammation occurred with ZnONP exposure. We also showed that p63 was consistently overexpressed due to ZnONP exposure in vivo and in vitro. This work provides unique findings on the p63 response and the pathobiology in response to ZnONPs, which could be important to the study of pulmonary toxicity and repair. HighlightsPulmonary effects caused by ZnONPs are determined in mice.Acute lung injury, lower airway inflammation, and p63 expression are observed.Pulmonary infiltration occurred by ZnONP at the secondary bronchial bifurcation.p63 was consistently over‐expressed at 24 h and 28 days after ZnONP exposure.

Effects of physical characteristics of carbon black on metabolic regulation in mice

Potential adverse effects of human exposure to carbon black (CB) have been reported, but limited knowledge regarding CB-regulated metabolism is currently available. To evaluate how physical parameters of CB influence metabolism, we investigated CB and diesel exhaust particles (DEPs) and attempted to relate various physical parameters, including the hydrodynamic diameter, zeta potential, and particle number concentrations, to lung energy metabolism in female BALB/c mice. A body weight increase was arrested by 3 months of exposure to CB of smaller-size fractions, which was negatively correlated with pyruvate in plasma. There were no significant differences in cytotoxic lactate dehydrogenase (LDH) or total protein in bronchoalveolar lavage fluid (BALF) after 3 months of CB exposure. However, we observed alterations in acetyl CoA and the NADP/NADPH ratio in lung tissues with CB exposure. Additionally, the NADP/NADPH ratio was associated with the zeta potential of CB. Mild peribronchiovascular and interstitial inflammation and multinucleated giant cells (macrophages) with a transparent and rhomboid appearance and containing foreign bodies were observed in lung sections. We suggest that physical characteristics of CB, such as the zeta potential, may disrupt metabolism after pulmonary exposure. These results, therefore, provide the first evidence of a link between pulmonary exposure to CB and metabolism.

Investigation into the pulmonary inflammopathology of exposure to nickel oxide nanoparticles in mice

We investigated the effects of nickel oxide nanoparticles (NiONPs) on the pulmonary inflammopathology. NiONPs were intratracheally installed into mice, and lung injury and inflammation were evaluated between 1 and 28 days. NiONPs caused significant increases in LDH, total protein, and IL-6 and a decrease in IL-10 in the BALF and increases in 8-OHdG and caspase-3 in lung tissues at 24 h. Airway inflammation was present in a dose-dependent manner from the upper to lower airways at 24 h of exposure as analyzed by SPECT. Lung parenchyma inflammation and small airway inflammation were observed by CT after NiONP exposure. 8-OHdG in lung tissues had increased with formation of fibrosis at 28 days. Focal adhesion was the most important pathways identified at 24 h as determined by protemics, whereas glutathione metabolism was the most important identified at 28 days. Our results demonstrated the pulmonary inflammopathology caused by NiONPs based on image-to-biochemical approaches.

Characterization of pulmonary protein profiles in response to zinc oxide nanoparticles in mice: a 24-hour and 28-day follow-up study

Although zinc oxide nanoparticles (ZnONPs) are recognized to cause systemic disorders, little is known about the mechanisms that underlie the time-dependent differences that occur after exposure. The objective of this study was to investigate the mechanistic differences at 24 hours and 28 days after the exposure of BALB/c mice to ZnONPs via intratracheal instillation. An isobaric tag for the relative and absolute quantitation coupled with liquid chromatography/tandem mass spectrometry was used to identify the differential protein expression, biological processes, molecular functions, and pathways. A total of 18 and 14 proteins displayed significant changes in the lung tissues at 24 hours and 28 days after exposure, respectively, with the most striking changes being observed for S100-A9 protein. Metabolic processes and catalytic activity were the main biological processes and molecular functions, respectively, in the responses at the 24-hour and 28-day follow-up times. The glycolysis/gluconeogenesis pathway was continuously downregulated from 24 hours to 28 days, whereas detoxification pathways were activated at the 28-day time-point after exposure. A comprehensive understanding of the potential time-dependent effects of exposure to ZnONPs was provided, which highlights the metabolic mechanisms that may be important in the responses to ZnONP.

Corrigendum: SUV39H1 Reduction Is Implicated in Abnormal Inflammation in COPD

This corrects the article DOI: 10.1038/srep46667.