Thesis (Ph. D.)--Lehigh University, 2004. / Includes bibliographical references and vita.

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Much of the aerosol research completed at the University of South Florida has revolved around evaluating industrial hygiene equipment and instrumentation in environmental chambers. Data collected during these studies has provided valuable baseline data on equipment and instrument performance under calm air conditions. A newly constructed wind tunnel now allows researchers to evaluate industrial hygiene instruments under moving air conditions. Because the wind tunnel is capable of producing wind velocities that a worker could encounter in the occupational setting, researchers may gain insight into instrument performance under simulated field conditions. Because aerosols can be introduced into the new wind tunnel testing section, researchers can also challenge industrial hygiene equipment and instrumentation with aerosols in sizes ranges that are of interest in public health. The purpose of this dissertation research was to develop a new wind tunnel to be used for aerosol research at the University of South Florida. Three specific aims had to be met for this study to be successful. They included: (1) designing a wind tunnel based on best practice information outlined in scientific literature, (2) constructing an operable wind tunnel to be used for aerosol research, and (3) characterizing wind tunnel performance by examining the wind tunnel velocity profile, turbulence intensity, and aerosol introduction/collection. The actual wind tunnel was constructed to a length of approximately 20 feet, a height of approximately 2 feet at its tallest point, and includes an entrance filter housing, a settling chamber, a contraction, a testing section, a diffuser, an exit filter housing, a fan, and exhaust duct. All components were designed and constructed using guidelines and best practices reported in the scientific literature. Velocity profile measurements were the first way that this wind tunnel was characterized. In order to successfully obtain measurements, the wind tunnel cross section was divided into 16 equal quadrants. Five measurements were taken for each quadrant at each wind velocity. Target wind velocities for this research were 0.5 m/s, 1.0 m/s, and 2.0 m/s. Actual average wind velocities of 0.48 m/s, 1.00 m/s, and 2.04 m/s. All were within established limits reported in the scientific literature. Turbulence intensity measurements were the second way that this wind tunnel was characterized. In order to successfully obtain measurements, the wind tunnel cross section was divided into 16 equal quadrants. Five measurements were taken for each quadrant at each wind velocity. Wind tunnels are typically designed to have the lowest turbulence intensity possible, generally below 10%. The overall average turbulence intensities for this wind tunnel at wind velocities of 0.5 m/s, 1.0 m/s, and 2.0 m/s were 9%, 10%, and 8% respectively. Overall turbulence intensity measurements were at or below 10%. Isokinetic sampling was the final method used to characterize this wind tunnel by collecting and detecting aerosols traveling through the wind tunnel testing section. The wind tunnel was operated at wind velocities of 0.5 m/s, 1.0 m/s, and 2.0 m/s with isokinetic sampling flow rates of 15.4 L/min, 30.9 L/min, and 61.7 L/min respectively. Monodisperse fluorescent polystyrene latex spheres were used as the test aerosol because they are uniform in size and shape and can be detected by fluorometry. The Blaustein Multi-Jet Atomizer (BLAM) was used to generate monodisperse fluorescent polystyrene latex aerosol 0.5 µm and 2.0 µm particles from liquid suspensions. The Vilnius Aerosol Generator (VAG) was used to generate monodisperse fluorescent polystyrene latex aerosol of 6.0 µm and 12.0 µm particles from dry powders. Nitrogen gas was used for delivering test aerosols into the wind tunnel. Five experimental runs were completed for each particle size and wind velocity for a total of 60 experimental runs. Fluorescence was detected in all 60 samples with average mass concentrations ranging from 0.000050 ng/ml to 0.002703 ng/ml. Based on velocity profile measurements, turbulence intensity measurements, and isokinetic sampling, the performance of University of South Florida wind tunnel was found to be excellent, indicating that it was designed and constructed appropriately. The wind tunnel can now successfully be used by researchers interested in evaluating industrial hygiene sampling equipment with aerosols ranging from 0.5 µm to 12.0 µm in moving air with velocities ranging from 0.5 m/s to 2.0 m/s.

Certain models of NIOSH-approved filtering facepiece air purifying respirators are manufactured with stapled head straps. Depending on the manufacturer, these head straps may be stapled to the filter media itself. This may cause leakage through the filter media of the respirator, potentially exposing the user to an unacceptable level of contaminant. In this study, monodisperse polystyrene latex (PSL) spheres were generated to challenge four replicates of a N95 single use respirator model made by the same manufacturer. Nominal particle sizes of the PSL spheres used to challenge the respirators were 0.5, 1, and 2 micrometers in diameter. All respirators were sealed onto a custom built testing assembly and tested in a sealed chamber. Particle sizes of interest were generated using a nebulizer, and passed through a diffusion dryer and a Krypton-85 radioactive source prior to entering the test chamber. The dryer reduces the humidity of the aerosol generating by the nebulizer, while the radioactive source neutralizes the charge of the aerosol cloud. The test chamber was constructed using a glass aquarium measuring 32 x 53 x 122 centimeters. Three stainless steel air diffusers were placed above the testing compartment to evenly distribute the aerosol in the chamber. An exhaust manifold was placed at the lower part of the chamber beneath another stainless steel diffuser below the area where test respirators were placed. The respirators were challenged as received from the manufacturer with 0.5, 1 and 2 micrometer-sized (PSL) spheres. The same procedure was repeated for each respirator after sealing the areas where the head straps were stapled with silicon rubber. Testing was conducted at a flow rate of 85 liters per minute, as specified in the NIOSH respirator testing protocol. A laser particle counter was used to measure the concentration inside and outside of the respirator. The results showed unsealed efficiencies for particle sizes 0.5, 1, and 2 micrometers of 96.68%, 99.72%, 99.88% and sealed efficiencies of 97.35%, 99.82%, 99.93% respectively. There were no differences for particle size or sealing at 1.0 and 2.0 micrometers. A significant drop in efficiency was observed when testing with 0.5 micrometer PSL spheres. The drops in efficiency are not sufficient to reduce the integrity of the respirator for N95 certification. However, the leakages detected will have a cumulative effect when added to other sources of single use respirator leakage in the field.

Surgical masks are intended to be used to prevent transmission of disease from a health care worker to a patient. Often times, they are relied upon by health care workers for their own protection. In light of recent developments regarding preparation for health care worker response to global infectious diseases such as H1N1 Influenza, health care workers may experience a false sense of security when wearing surgical masks. The goal of this study was to evaluate the filtration efficiency of a double strap tie-on surgical mask. The manufacturer asserts a >95% efficiency with a 0.1 um challenge aerosol under FDA testing procedures. The NIOSH Title 42 CFR Part 84 certification criteria call for testing at a rate of 85 lpm representing a human moderate to heavy work load breathing rate. Three sizes of monodispersed aerosols (polystyrene latex beads: 0.5 um, 1.0 um, 2.0 um) were used. The specific aims were to measure the collection efficiencies of this mask for the various particle sizes. Two tests were performed. In the first, masks were affixed to a dummy head and the edges of the mask were not sealed. In the second, the edges of the masks were sealed to the head using silicone sealant, so all penetration was through the filtering material of the mask. Differences in upstream and downstream particle concentrations were measured. Thus, penetration by leakage around the mask and through the filtering material was measured. The experimental set up involved passing the aerosol from the nebulizer through a diffusion dryer and Kr-85 charge equilibrator ensuring a dry charge neutralized aerosol cloud for detection by a LASAIR particle counter. The analysis revealed that the filtration efficiency for 0.5 um particles ranged from 3% to 43% for the unsealed masks and 42% to 51% for the sealed. For 1.0 um particles, the efficiency was 58% to 75% for unsealed and 71% to 84% for sealed masks. For 2.0 um, the efficiency was 58% to 79% for unsealed masks and 69% to 85% for the sealed masks. The data were statistically significant and indicated that surgical masks were associated with very low filtration efficiency. This suggests that they may be inadequate against airborne viruses and bacteria.

Yes / Nanoparticles are increasingly used in industry and medicine due to their unique physiochemical properties such as their small size, charge, shape, chemical architecture, large surface area, surface reactivity and media interactions, etc [1-5]. However, very little is still known on the interactions between nanoparticles and the biological system. This study aims to evaluate the cytotoxicity of polystyrene latex bead nanoparticles on HaCat and HeLa cell lines. Carboxyl-modified 20 nm polystyrene NPs core labelled with fluorophore were from Invitogen. We chose to use polystyrene NPs because this specific type of NP is being increasingly characterized for use in nanosensors and drug nanocarrier investigations. 1x 104 cells/100 μl of cell culture medium were plated into 96-well plates in triplicate, measuring activity post 24 hours at concentrations of 10, 50, 100 μg/ml of polystyrene NPs exposure. The extracellular lactate dehydrogenase release was measured by using a colorimetric CytoTox 96 nonradioactive assay kit from Promega and the absorbance were recorded at 450nm (FLUO-star) with Elisa micro plate reader. The MTT assay was used to evaluate mitochondrial activity. This was performed by inserting a premixed optimized dye solution in the culture wells. The Absorbance was recorded at 570 nm, from the recorded absorbance is directly proportional to the number of live cells. The cell lines were kept in a plastic T-75cm2 tissue culture flasks grown in DMEM. We found that cytotoxicity of polystyrene NPs on both cells was concentration dependent. For Hela cells, with exporesure of polystyrene NPs at concentrations of 10, 50, 100 μg/ml for 24 hrs, the percentage cytotoxicity of positive control for LDH assay was 35.9%, 49.5% and 73.4% respectively. With the MTT cell viability assay the percentage MTT reduction of negative control was 88.9%, 42.9% and 26.4% respectively. Cell toxicity increased with increasing polystyrene NPs concentration. For HaCaT cells, the cytotoxic effect is less significant than those on Hela cells. With MTT assay, when compared to HaCaT cells exposed to a negative control containing only PBS, the cell viability decreased as the concentrations of NPs increased. Cells exposed to 100μg/ml of polystyrene NPs for a period of 24 hours compared to those exposed to a positive control (100% cell viability) had an average cell viability of 49%, with those numbers decreasing from 59% for cells exposed to 10μg/ml of polystyrene NPs to 57% for cells exposed to 50μg/ml of polystyrene NPs. Our results indicated that polystyrene NPs acted differently in two different cell types and that cautions should be taken about its cytotoxicity. Further understanding of the mechanism involving the ROS generation could provide more information on how polystyrene NPs increase cytotoxicity.

No / Microcarrier cell culture systems provide an attractive alternative to the conventional monolayer cell culture for cell amplification, due to their high surface area-to-volume ratio. Unlike enzymatic methods for removing cells from microcarriers after cell culture, which can lead to irreversible damage of the cells, microcarriers which release cells by temperature adjustment have been developed. This was achieved by grafting a temperature-responsive polymer, poly(N-isopropylacrylamide) (PNIPAAm), on the microcarrier surface. This review comprehensively presents various methods to prepare such thermo-responsive microcarriers based on PNIPAAm. These methods include the grafting-to technique, grafting-from technique, grafting-through technique, along with methods leading to PNIPAAm hydrogel beads, seeded polymerization, and non-covalent adsorption. The methods for controlling PNIPAAm grafting density, molecular weight and molecular architecture are also outlined. Further, the efficiency of cell attachment, proliferation and thermally-induced detachment of such thermo-responsive microcarriers is introduced and compared. (C) 2015 Elsevier Ltd. All rights reserved.