Toxicity assessment of CeO2 and CuO nanoparticles at the air-liquid interface using bioinspired condensational particle growth.

CeO2 and CuO nanoparticles (NPs) are used as additives in petrodiesel to enhance engine performance leading to reduced diesel combustion emissions. Despite their benefits, the additive application poses human health concerns by releasing inhalable NPs into the ambient air. In this study, a bioinspired lung cell exposure system, Dosimetric Aerosol in Vitro Inhalation Device (DAVID), was employed for evaluating the toxicity of aerosolized CeO2 and CuO NPs with a short duration of exposure (≤10 min vs. hours in other systems) and without exerting toxicity from non-NP factors. Human epithelial A549 lung cells were cultured and maintained within DAVID at the air-liquid interface (ALI), onto which aerosolized NPs were deposited, and experiments in submerged cells were used for comparison. Exposure of the cells to the CeO2 NPs did not result in detectable IL-8 release, nor did it produce a significant reduction in cell viability based on lactate dehydrogenase (LDH) assay, with a marginal decrease (10%) at the dose of 388 µg/cm2 (273 cm2/cm2). In contrast, exposure to CuO NPs resulted in a concentration dependent reduction in LDH release based on LDH leakage, with 38% reduction in viability at the highest dose of 52 µg/cm2 (28.3 cm2/cm2). Cells exposed to CuO NPs resulted in a dose dependent cellular membrane toxicity and expressed IL-8 secretion at a global dose five times lower than cells exposed under submerged conditions. However, when comparing the ALI results at the local cellular dose of CuO NPs to the submerged results, the IL-8 secretion was similar. In this study, we demonstrated DAVID as a new exposure tool that helps evaluate aerosol toxicity in simulated lung environment. Our results also highlight the necessity in choosing the right assay endpoints for the given exposure scenario, e.g., LDH for ALI and Deep Blue for submerged conditions for cell viability.

Efficient adsorption of aromatic and aliphatic hydrocarbons by electrospun hydrophobic PTFE-NiO composite nanofiber filter mats.

Aromatic and aliphatic hydrocarbons (AAHs) are comprised of a variety of gaseous chemicals that may affect human and environmental health. To remove AAHs from air, polytetrafluoroethylene-nickel oxide (PTFE-NiO) composite nanofiber filter mats (NFMs) were synthesized and characterized for their ability to effectively adsorb AAHs. The NiO-nanoparticle-doped mats were fabricated by green electrospinning of PTFE and polyvinyl alcohol (PVA) mixtures added with nickel (II) nitrate hexahydrate in the spinning solution followed by surface heat treatment. FE-SEM FTIR, Raman spectroscopy, sessile drop and Jar methods were applied as characterization techniques. The diameter of the electrospun nanofibers without NiO dopant ranged from 0.34 ± 21.61 to 0.23 ± 10.12 µm, whereas a reduction in diameter of NiO-doped nanofibers was obtained, ranging between pristine to 0.25 ± 24.12 µm and 0.12 ± 85.75 µm with heat treatment. 6% (by weight) NiO-doped PTFE composite NFMs exhibited a high water-contact angle of 120 ± 2.20 degrees; the high hydrophobicity value aided self-cleansing property of NFMs for practical applications. UV adsorption capability for heat-treated PTFE-NiO NFMs was evaluated for three AAHs, and the results showed that 6 wt% NiO adsorbed 1.41, 0.67, and 0.73 µg/mg of toluene, formaldehyde and acetone, respectively. These findings reveal the potential applicability of the prepared filter mats for capturing various AAHs from polluted air.

Integration of sample preparation with RNA-Amplification in a hand-held device for airborne virus detection.

Aerosol transmission is one of the three major transmission routes of respiratory viruses. However, the dynamics and significance of the aerosol transmission route are not well understood, partially due to the lack of rapid and efficient tools for on-the-spot detection of airborne viruses. We report a hand-held device that integrates a 3D-printed sample preparation unit with a laminated paper-based RNA amplification unit. The sample preparation unit features an innovative reagent delivery scheme based on a ball-based valve capable of storing and delivering reagents through the rotation of the unit without manual pipetting, while the paper-based unit enables RNA enrichment and reverse transcription loop-mediated isothermal amplification (RT-LAMP). We have determined the detection limit of the integrated sample-preparation/amplification device (SPAD) at 1 TCID50 H1N1 influenza viruses in 140 µL aqueous sample. Further, we integrated SPAD with a previously reported viable virus aerosol sampler (VIVAS), a water-vapor-based condensational growth system capable of collecting aerosolized virus particles (Pan et al., 2016) [1]. Using the combined VIVAS-SPAD platform, we have demonstrated the collection/detection of lab-generated, airborne H1N1 influenza viruses in 65 min, suggesting that the platform has a potential for detecting and monitoring airborne virus transmission during outbreaks. The effective sampling and rapid detection of airborne viruses by the sample-to-answer platform will also help us better understand the dynamics and significance of aerosol transmission of infectious disease.

Viable SARS-CoV-2 in the air of a hospital room with COVID-19 patients.

OBJECTIVES: Because the detection of SARS-CoV-2 RNA in aerosols but failure to isolate viable (infectious) virus are commonly reported, there is substantial controversy whether severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) can be transmitted through aerosols. This conundrum occurs because common air samplers can inactivate virions through their harsh collection processes. We sought to resolve the question whether viable SARS-CoV-2 can occur in aerosols using VIVAS air samplers that operate on a gentle water vapor condensation principle. METHODS: Air samples collected in the hospital room of two coronavirus disease-2019 (COVID-19) patients, one ready for discharge and the other newly admitted, were subjected to RT-qPCR and virus culture. The genomes of the SARS-CoV-2 collected from the air and isolated in cell culture were sequenced. RESULTS: Viable SARS-CoV-2 was isolated from air samples collected 2 to 4.8 m away from the patients. The genome sequence of the SARS-CoV-2 strain isolated from the material collected by the air samplers was identical to that isolated from the newly admitted patient. Estimates of viable viral concentrations ranged from 6 to 74 TCID50 units/L of air. CONCLUSIONS: Patients with respiratory manifestations of COVID-19 produce aerosols in the absence of aerosol-generating procedures that contain viable SARS-CoV-2, and these aerosols may serve as a source of transmission of the virus.

Viable SARS-CoV-2 in the air of a hospital room with COVID-19 patients.

Background - There currently is substantial controversy about the role played by SARS-CoV-2 in aerosols in disease transmission, due in part to detections of viral RNA but failures to isolate viable virus from clinically generated aerosols. Methods - Air samples were collected in the room of two COVID-19 patients, one of whom had an active respiratory infection with a nasopharyngeal (NP) swab positive for SARS-CoV-2 by RT-qPCR. By using VIVAS air samplers that operate on a gentle water-vapor condensation principle, material was collected from room air and subjected to RT-qPCR and virus culture. The genomes of the SARS-CoV-2 collected from the air and of virus isolated in cell culture from air sampling and from a NP swab from a newly admitted patient in the room were sequenced. Findings - Viable virus was isolated from air samples collected 2 to 4.8m away from the patients. The genome sequence of the SARS-CoV-2 strain isolated from the material collected by the air samplers was identical to that isolated from the NP swab from the patient with an active infection. Estimates of viable viral concentrations ranged from 6 to 74 TCID50 units/L of air. Interpretation - Patients with respiratory manifestations of COVID-19 produce aerosols in the absence of aerosol-generating procedures that contain viable SARS-CoV-2, and these aerosols may serve as a source of transmission of the virus.

Mimicking the human respiratory system: Online in vitro cell exposure for toxicity assessment of welding fume aerosol.

In assessing the biological impact of airborne particles in vitro, air-liquid interface (ALI) exposure chambers are increasingly preferred over classical submerged exposure techniques, albeit historically limited by their inability to deliver sufficient aerosolized dose. A novel ALI system, the Dosimetric Aerosol in Vitro Inhalation Device (DAVID), bioinspired by the human respiratory system, uses water-based condensation for highly efficient aerosol deposition to ALI cell culture. Here, welding fumes (well-studied and inherently toxic ultrafine particles) were used to assess the ability of DAVID to generate toxicological responses between differing welding conditions. After fume exposure, ALI-cultured cells showed reductions in viability that were both distinct between welding conditions and linearly dose-dependent with respect to exposure time; comparatively, submerged cell cultures ran in parallel did not show these trends across exposure levels. DAVID delivers a substantial dose in minutes (> 100 µg/cm2), making it preferable over previous ALI systems, which require hours of exposure to deliver sufficient dose, and over submerged techniques, which lack comparable physiological relevance. DAVID has the potential to provide the most accurate assessment of in vitro toxicity yet from the perspectives of physiological relevance to the human respiratory system and efficiency in collecting ultrafine aerosol common to hazardous exposure conditions.

In Vitro Aerosol Exposure to Nanomaterials: From Laboratory to Environmental Field Toxicity Testing.

Exposure to nanomaterials (NMs) is inevitable, requiring robust toxicological assessment to understand potential environmental and human health effects. NMs are favored in many applications because of their small size; however, this allows them to easily aerosolize and, subsequently, expose humans via inhalation. Toxicological assessment of NMs by conventional methods in submerged cell culture is not a relevant way to assess inhalation toxicity of NMs because of particle interference with bioassays and changes in particokinetics when dispersed in medium. Therefore, an in vitro aerosol exposure chamber (AEC) was custom designed and used for direct deposition of NMs from aerosols in the environment to the air-liquid interface of lung cells. Human epithelial lung cell line, A549, was used to assess the toxicity of copper, nickel, and zinc oxide nanopowders aerosolized by acoustic agitation in laboratory study. Post optimization, the AEC was used in the field to expose the A549 cells to NM aerosols generated from firing a hand gun and rifle. Toxicity was assessed using nondestructive assays for cell viability and inflammatory response, comparing the biologic effect to the delivered mass dose measured by inductively coupled plasma-mass spectrometry. The nanopowder exposure to submerged and ALI cells resulted in dose-dependent toxicity. In the field, weapon exhaust from the M4 reduced cell viability greater than the M9, while the M9 stimulated inflammatory cytokine release of IL-8. This study highlights the use of a portable chamber with the capability to assess toxicity of NM aerosols exposed to air-liquid interface in vitro lung cell culture.

Condensational particle growth device for reliable cell exposure at the air-liquid interface to nanoparticles.

A first-of-its-kind aerosol exposure device for toxicity testing, referred to as the Dosimetric Aerosol in Vitro Inhalation Device (DAVID), was evaluated for its ability to deliver airborne nanoparticles to lung cells grown as air-liquid interface (ALI) cultures. For inhalation studies, ALI lung cell cultures exposed to airborne nanoparticles have more relevancy than the same cells exposed in submerged culture because ALI culture better represents the respiratory physiology and consequently more closely reflect cellular response to aerosol exposure. In DAVID, water condensation grows particles as small as 5 nm to droplets sized > 5 µm for inertial deposition at low flow rates. The application of DAVID for nanotoxicity analysis was evaluated by measuring the amount and variability in the deposition of uranine nanoparticles and then assessing the viability of ALI cell cultures exposed to clean-air under the same operational conditions. The results showed a low coefficient of variation, < 0.25, at most conditions, and low variability in deposition between the exposure wells, trials, and operational flow rates. At an operational flow rate of 4 LPM, no significant changes in cell viability were observed, and minimal effects observed at 6 LPM. The reliable and gentle deposition mechanism of DAVID makes it advantageous for nanoparticle exposure.

Performance of silver, zinc, and iron nanoparticles-doped cotton filters against airborne E. coli to minimize bioaerosol exposure.

To overcome limitations of existing air-cleaning filters in capturing and deactivating aerosolized microorganisms, this study was embarked to evaluate novel Ag, Zn, and Fe nanoparticle-doped cotton filters (AgCt, ZnCt, FeCt), as biocidal filters for bioaerosol attenuation. To evaluate the biocidal activity of the nanocomposite filters, the survival of lab-generated E. coli after collection on each filter material was compared to collection on an undoped cotton control filter and in a BioSampler. Relative humidity (RH) affected the survival of bacteria on the filters, and the optimal RH was found to be 50 ± 5%. The physical removal efficiency (PRE) determined by an optical particle counter was 99.9 ± 0.7% for ZnCt, 97.4 ± 1.2% for AgCt, and 97.3 ± 0.6% for FeCt, where the control showed only 77.4 ± 6.3% for particles > 500 nm. The doped filters showed 100% viable removal efficiency (VRE). Importantly, the VRE of the nanocomposite filters after four cycles remained nearly 99% and was greater than the cotton control filter at 76.6 ± 3.2%. Adding to its benefits, the AgCt filters had a lower pressure drop than the FeCt and ZnCt filters and the cotton control. The permeability for the cotton control filter was 3.38 × 10-11 m2 while that for the AgCt filter was slightly higher (3.64 × 10-11 m2) than the other filters as well. Overall, these results suggest that nanocomposite-doped filter media, particularly AgCt, can provide effective protection against airborne pathogens with a lower pressure drop, elevated collection efficiency, and better disinfection capability as compared to untreated cotton filters, which are all important features for practical biocidal applications. Graphical abstract.

Characterization of exposure to byproducts from firing lead-free frangible ammunition in an enclosed, ventilated firing range.

U.S. Air Force small arms firing ranges began using copper-based, lead-free frangible ammunition in the early 2000s due to environmental and health concerns related to the use of lead-based ammunition. Exposure assessments at these firing ranges have routinely detected chemicals and metals in amounts much lower than their mass-based occupational exposure limits, yet, instructors report work-related health concerns including respiratory distress, nausea, and headache. The objective of this study at one firing range was to characterize the aerosol emissions produced by weapons during firing events and evaluate the ventilation system's effectiveness in controlling instructor exposure to these emissions. The ventilation system was assessed by measuring the range static air pressure differential and the air velocity at the firing line. Air flow patterns were near the firing line. Instructor exposure was sampled using a filter-based air sampling method for metals and a wearable, real-time ultrafine particle counter. Area air sampling was simultaneously performed to characterize the particle size distribution, morphology, and composition. In the instructor's breathing zone, the airborne mass concentration of copper was low (range = <1 µg/m3 to 16 µg/m3), yet the ultrafine (nanoscale) particle number concentration increased substantially during each firing event. Ultrafine particles contained some copper and were complex in morphology and composition. The ventilation assessment found that the average velocity across all shooting lanes was acceptable compared to the recommended guideline (20% of the ideal 0.38 m/s (75 ft/min). However, uniform, downrange airflow pattern requirements were not met. These results suggest that the mass-based occupational exposure limits, as applied to this environment, may not be protective enough to eliminate health complaints reported by instructors whose full-time job involves training personnel on weapons that fire lead-free frangible ammunition. Using an ultrafine particle counter appears to be an alternative method of assessing ventilation effectiveness in removing ultrafine particulate produced during firing events.