Abstract  Objective: The objectives are to develop inhalation dosimetry models of the flavoring agents diacetyl, 2, 3-pentanedione, and acetoin to predict uptake throughout the rat and human respiratory tracts and use the results with histopathology data from 2-week, nose-only inhalation exposures in Sprague-Dawley rats to assess relationships between predicted dose and in vivo responses. Methods: Computational fluid dynamics (CFD) models of the nasal passages were used to simulate inspiratory airflow and vapor uptake and mechanistic models of the lung airways were used to simulate vapor uptake during a breathing cycle. Results: Diacetyl and 2, 3-pentanedione demonstrated similar uptake and wall mass flux patterns throughout the respiratory tract. Acetoin, being more soluble, was rapidly absorbed in the nasal and upper lung airways. At a 10 ppm exposure concentration and resting breathing conditions, nasal uptake of diacetyl, 2, 3-pentanedione, and acetoin was 30.9, 30.3, and 73.6% in the rat, and 8.7, 9.3, and 32.5% in the human, respectively; total respiratory tract uptake was 76.5, 76.8, and 93.0% in the rat and 79.6, 81.1, and 85.9% in the human, respectively. Wall mass flux patterns aligned with previously reported in vivo observations of histopathological effects in the rat respiratory tract following 8.75, 17.5, or 35 ppm diacetyl or 2, 3-pentanedione exposure and can be used to evaluate dose-response behavior. Conclusions: Dose-response assessment of inhaled vapors demonstrates the utility of dosimetry models for interspecies extrapolation and chemical comparisons and how their use is an important part of risk characterization as non-animal alternatives are more widely considered.

Abstract  An in vitro study was conducted with the goal of developing empirical correlations to predict the deposition of particles with an aerodynamic diameter of 0.5-5.3 mu m in nasal airways of children with ages 4-14 years. CT images of nasal airways of thirteen healthy subjects and one with congested nasal airways were used for fabricating the replicas by rapid prototyping. Replicas included nasal airways to the level of the upper trachea as well as the face. Four physiological breathing patterns (sinusoidal waves) of children were simulated by a pulmonary waveform generator. Using an Electrical Low Pressure Impactor (ELPI) we measured deposition of sunflower oil particles generated by a Collison atomizer. Moreover, using the same setup, particle deposition in five adult replicas fabricated using MRI images was measured for direct comparison with the child replicas and in vivo data available for adults. Deposition increased by increasing particle size and flow rate, indicating impaction as the dominant deposition mechanism. Existing correlations for adults were unable to reduce the scatter in the deposition data for children. A correlation was developed for prediction of deposition including the relevant non-dimensional numbers (Stokes and Reynolds numbers) that were calculated considering the dimensions of the airways. The corrected correlations should be useful for exposure estimation of children and for efficient pediatric drug delivery using face masks. (C) 2011 Elsevier Ltd. All rights reserved.

Abstract  Increased use of electronic cigarette (EC) products has called for studying the health consequences from these new products. Government agencies such as the FDA are interested in toxicological, exposure data and other relevant information as a part of regulating the use of the products. A key step in risk assessment of EC products is to determine the inhaled dose of various constituents in the EC aerosol. Given the challenges in collecting human and animal data, mathematical models may guide and complement data collection. Models developed for combustible tobacco smoke may be improved and extended to EC aerosol to allow for higher volatility and hygroscopicity of the latter. Both these properties greatly affect the dynamics and deposition of inhaled aerosol. In this study, we developed a deposition model for EC aerosol based on realistic vaping scenarios. The model included several steps representing puff withdrawal into the oral cavity, mouth hold, dilution of the EC puff in the mouth with the subsequent dilution from inhaled air, inhalation of the puff into the lower respiratory tract, lung-hold, and exhalation. Results for deposition of individual constituents and droplet dynamics within the oral cavity of the respiratory tract will be presented in this publication. The model accounted for thermodynamic interactions between the droplet and vapor phases of each constituent in the aerosol mixture. Deposition from droplets and uptake of vapor constituents were calculated. The deposited mass fraction for each vapor constituent and droplets were also calculated based on the total mass of the inhaled constituents. The inhalation model developed in this study could help the academics, government agencies and tobacco manufacturers with development of risk models for EC use.

Abstract  "Standard saccharine test is used to detect nasal mucociliary clearance time in healthy individuals both adults and children. The length of the nose was measured radiologically and with the help of a soft malleable rubber catheter. For healthy individuals, mean nasal mucociliary clearance lime is 8.2 minutes in children and 9.5 minutes in adults. The mean nasal mucociliary clearance rates were 11.1 mm/min for children and 12.7 mm/min for adults.Deviated nasal septum, chronic sinusitis, allergic rhinitis, atrophic rhinitis, chronic smokers and patients with recent nasal packings were taken as diseased conditions in adults, whereas children with adenoid hyperplasia were taken for the study. In all of these, nasal mucociliary clearance was significantly prolonged."

Abstract  Traditionally, empirical correlations for predicting respiratory tract deposition of inhaled aerosols have been developed using limited available in vivo data. More recently, advances in medical image segmentation and additive manufacturing processes have allowed researchers to conduct extensive in vitro deposition experiments in realistic replicas of the upper and central branching airways. This work has led to a collection of empirical equations for predicting regional aerosol deposition, especially in the upper, nasal and oral airways. The present section reviews empirical correlations based on both in vivo and in vitro data, which may be used to predict total and regional deposition. Equations are presented for predicting total respiratory deposition fraction, mouth-throat fraction, nasal, and nose-throat fractions for a large variety of aerosol sizes, subject age groups, and breathing maneuvers. Use of these correlations to estimate total lung deposition is also described.

Abstract  Guinea pigs have been used as a surrogate for humans in a number of inhalation toxicology studies by exposing the animals to airborne viruses, biological agents, and other chemicals. Findings from these studies can be extrapolated to humans based on a particular dose-metric of interest to determine potential health effects in humans. In the absence of exposure-dose data, mathematical models can be developed for the transport and deposition of inhaled particles in the lungs of guinea pigs to allow for predictions of lobar, regional, and local deposition of particles.

Abstract  To facilitate computational toxicology, we developed an approach for generating high-resolution lung-anatomy and particle-deposition mouse models. Major processing steps of our method include mouse preparation, serial block-face cryomicrotome imaging, and highly automated image analysis for generating three-dimensional (3D) mesh-based models and volume-based models of lung anatomy (airways, lobes, sublobes, and near-acini structures) that are linked to local particle-deposition measurements. Analysis resulted in 34 mouse models covering 4 different mouse strains (B6C3F1: 8, BALB/C: 11, C57Bl/6: 8, and CD-1: 7) as well as both sexes (16 male and 18 female) and different particle sizes [2 mu m (n = 15), 1 mu m (n = 16), and 0.5 mu m (n = 3)]. On average, resulting mouse airway models had 1,616.9 +/- 298.1 segments, a centerline length of 597.6 +/- 59.8 mm, and 1,968.9 +/- 296.3 outlet regions. In addition to 3D geometric lung models, matching detailed relative particle-deposition measurements are provided. All data sets are available online in the lapdMouse archive for download. The presented approach enables linking relative particle deposition to anatomical structures like airways. This will in turn improve the understanding of site-specific airflows and how they affect drug, environmental, or biological aerosol deposition. NEW & NOTEWORTHY Computer simulations of particle deposition in mouse lungs play an important role in computational toxicology. Until now, a limiting factor was the lack of high-resolution mouse lung models and measured local particle-deposition information, which are required for developing accurate modeling approaches (e.g., computational fluid dynamics). With the developed imaging and analysis approach, we address this issue and provide all of the raw and processed data in a publicly accessible repository.

Abstract  ObjectiveCigarette smoking can lead to a host of adverse health effects such as lung and heart disease. Increased lung cancer risk is associated with inhalation of carcinogens present in a puff of smoke. These carcinogenic compounds deposit in the lung at different sites and trigger a cascade of events leading to adverse outcomes. Understanding the site-specific deposition of various smoke constituents will inform the study of respiratory diseases from cigarette smoking. We previously developed a deposition model for inhalation of aerosol from electronic nicotine delivery systems. In this study, the model was modified to simulate inhalation of cigarette smoke consisting of soluble and insoluble tar, nicotine, and cigarette-specific constituents that are known or possible human carcinogens.Materials and MethodsThe deposition model was further modified to account for nicotine protonation and other cigarette-specific physics-based mechanisms that affect smoke deposition. Model predictions showed a total respiratory tract uptake in the lung for formaldehyde (99%), nicotine (80%), and benzo[a]pyrene (60%).ResultsThe site of deposition and uptake depended primarily on the constituent's saturation vapor pressure. High vapor pressure constituents such as formaldehyde were preferentially absorbed in the oral cavity and proximal lung regions, while low vapor pressure constituents such as benzo[a]pyrene were deposited in the deep lung regions. Model predictions of exhaled droplet size, droplet retention, nicotine retention, and uptake of aldehydes compared favorably with experimental data.ConclusionThe deposition model can be integrated into exposure assessments and other studies that evaluate potential adverse health effects from cigarette smoking.

Abstract  Evaluation of the regional intranasal delivery of locally acting drugs in children is challenging. Anatomical nasal airway replicas potentially can provide a robust pre -clinical tool to test the performance of devices and formulations. However, there is often a challenge in identifying the nasal geometries that can reasonably be indicative of in vivo regional mass distribution of administered drug. This in vitro study was designed to investigate the regional intranasal drug delivery in 20 children, 2 -11 years old (50% 2 -6 years old and 50% female), using two commercially available suspension nasal spray products with different nozzle designs, plume characteristics, and active pharmaceutical ingredients. High -resolution computed tomography scans of the sinonasal region of pediatric human subjects with healthy nasal airways, reviewed and scored by a head and neck surgeon, were used to develop 20 three-dimensional (3D) replicas of the nasal airways. The 3D replicas were segmented into the two regions: anterior and posterior to the internal nasal valve (INV). They were then rapid prototyped in high clarity rigid plastic (Accura ClearVue). Each side of the septum of the 20 subjects was examined separately, resulting in 40 singular nasal cavities. A nozzle -specific spray tip holder was designed for each case to ensure consistent administration (insertion length, sagittal angle, and coronal angle) in all replicates. The wide range of posterior drug delivery observed in the forty geometries indicated significant intersubject variability in pediatric intranasal drug delivery. Three nasal geometries representing low, medium, and high levels of drug delivery to the target region, posterior to the INV, were chosen from the 40 nasal cavities. Our vision is that these three nasal geometries can potentially be beneficial in determining whether performance differences between test and reference nasal spray products are present that may affect their bioequivalence in children. They also may be useful when applied in parallel with similar adult nasal geometries, previously developed following a similar procedure, to provide additional insights into pediatric nasal drug delivery with innovator products in children in lieu of extending clinical studies to include pediatric subjects.

Abstract  Risk of lung damage from inhaled chemicals or substances has long been assessed using animal models. However, New Approach Methodologies (NAMs) that replace, reduce, and/or refine the use of animals in safety testing such as 2D and 3D cultures are increasingly being used to understand human-relevant toxicity responses and for the assessment of hazard identification. Here we review 2D and 3D lung models in terms of their application for inhalation toxicity assessment. We highlight a key case study for the Organization for Economic Cooperation and Development (OECD), in which a 3D model was used to assess human toxicity and replace the requirement for a 90-day inhalation toxicity study in rats. Finally, we consider the regulatory guidelines for the application of NAMs and potential use of different lung models for aerosol toxicity studies depending on the regulatory requirement/context of use.

Abstract  Quantifying drug delivery to the site of action using locally-acting nasal suspension sprays is a challenging but important step toward understanding bioequivalence (BE) between test and reference products. The main objective of this study was to investigate the in vitro deposition pattern of two common but different locally-acting nasal suspension sprays using multiple nasal cavities. Twenty anatomically accurate nasal replicas were developed from high-resolution sinonasal computed tomography scans of adults with healthy nasal airways. The airways were segmented into two regions of anterior and posterior to the internal nasal valve. Both sides of the septum were considered separately; hence, 40 nasal cavities were studied. The positioning of the spray nozzle in all 40 cavities was characterized by the head angle, coronal angle, and the insertion depth. Despite using a controlled protocol to minimize the anterior losses, a wide range of variability in posterior drug delivery was observed. The observed intersubject variability using this in vitro method may have important implications for understanding BE of locally-acting nasal suspension sprays.

Abstract  Respiratory tract dosimetry predictions for inhalation of tobacco product smoke and aerosols are sensitive to the values of the physicochemical properties of constituents that make up the puff. Physicochemical property values may change significantly with temperature, particularly in the oral cavity and upper airways of the lung, where the puff undergoes adjustments from high temperatures in the tobacco product to reach body temperature. The assumption of fixed property values may introduce uncertainties in the predicted doses in these and other airways of the lung. To obtain a bound for the uncertainties and improve dose predictions, we studied temperature evolution of the inhaled puff in the human respiratory tract during different puff inhalation events. Energy equations were developed for the transport of the puff in the respiratory tract and were solved to find air and droplet temperatures throughout the respiratory tract during two puffing scenarios: 1. direct inhalation of the puff into the lung with no pause in the oral cavity, and 2. puff withdrawal, mouth hold, and puff delivery to the lung via inhalation of dilution air. These puffing scenarios correspond to the majority of smoking scenarios. Model predictions showed that temperature effects were most significant during puff withdrawal. Otherwise, the puff reached thermal equilibrium with the body. Findings from this study will improve predictions of deposition and uptake of puff constituents, and therefore inform inhalation risk assessment from use of electronic nicotine delivery systems (ENDS) and combusted cigarettes.

Abstract  Nine oral airway replicas of children 6-14 years old were used for measuring deposition of 0.5-6.3 mu m diameter particles during inhalation of four representative (sinusoidal) breathing patterns typical of those observed during the use of nebulizers by children. For a given particle size and inhalation flow rate, deposition in the present child replicas is higher than in adults and is not well predicted by adult correlations, even when age-related scaling factors are applied. However, by using subject-specific geometrical dimensions of the airway replicas, dimensionless correlations that predict deposition of micrometer-sized particles in children's oral airways during tidal breathing are presented. Such best fitting correlation gives deposition efficiency eta(2) = [1-1/(4.99(stk Re-2.41(-0.17))+1)] x 100, where Re and Stk are Reynolds and Stokes numbers with length scale given by V/As where V is airway volume and A(s) is airway surface area. (C) 2012 Elsevier Ltd. All rights reserved.

Abstract  Experimentally measured deposition of ultrafine particles, ranging from 13-100 nm in diameter, in nasal airway replicas of ten infants aged 3-18 months is presented. The replicas included the face, nostrils, and nasal airways including the upper trachea. A differential mobility analyzer (DMA) and a condensation particle counter (CPC) were used to quantify the nasal deposition by comparing the number of polydisperse sodium chloride particles, generated by evaporation from a Collison atomizer, at the inlet and outlet of the replicas. Particles were individually classified in size by DMA and subsequently were counted one size bin at a time by CPC upstream and downstream of each replica. Since in vivo data is not available for infants to compare to, we validated our experimental procedure instead by comparing deposition in nasal airway replicas of six adults with in vivo measurements reported in literature. In the infant replicas, tidal inhalation was simulated at two physiologically compatible flow rates and the effect of flow rate on deposition was found to be small. Deposition obtained at constant flow rates is lower than with tidal breathing, indicating the importance of unsteadiness, in contrast to similar data in adults where unsteadiness is known to be unimportant. An empirical equation, containing geometrical features of the nasal airways in the form of related non-dimensional dynamical parameters (Reynolds, Schmidt, and Womersley numbers), was best fitted to the infant data. This equation may be useful for a priori prediction of nasal deposition and intersubject variability during exposure of infants to ultrafine aerosols.

Abstract  Characterization of internal dose and potential health impact of inhaling aerosol from an Electronic Nicotine Delivery System (ENDS) requires understanding and estimation of the regional deposition and absorption of the aerosol constituents in the respiratory tract. The aerosol generated from ENDS is a highly unstable mixture of multi-constituent aerosols and their vapor constituents. The mixture undergoes rapid changes once inhaled into the respiratory tract. Measurement of the deposited dose is a formidable challenge and no reliable method is currently available. Hence, a model for the deposition of aerosol components and vapor constituent of an ENDS aerosol mixture was developed based on previously constructed models for particle deposition and vapor uptake in the respiratory tract. Constituent phase instability and rapid mass exchange within the aerosol mixture and with airway walls required the coupling of the aerosol and vapor phases for all constituents. The fate of the ENDS aerosol mixture was determined throughout the respiratory tract for a typical vaping scenario consisting of puff withdrawal, mouth hold, mixing of the puff with dilution air at the end of mouth hold, lung inhalation, lung hold, and exhalation. Model predictions indicated that over 90% of constituents with medium vapor pressure (e.g., nicotine and propylene glycol or PG) were delivered to the lung tissues by both aerosol deposition and vapor uptake, which occurred in all regions of respiratory tract. Low vapor pressure constituents (e.g., glycerin) mostly remained in the aerosols and were delivered to the lung by aerosol deposition alone. The dosimetry model is a useful tool to estimate the internal exposure to the constituents in the ENDS aerosol and can provide valuable insights for risk assessments.

Abstract  Aerosol dosimetry estimates for mouse strains used as models for human disease are not available, primarily because of the lack of tracheobronchial airway morphometry data. By using micro-CT scans of in-situ prepared lung casts, tracheobronchial airway morphometry for four strains of mice were obtained: Balb/c, AJ, C57BL/6, and Apoe(-/-). The automated tracheobronchial airway morphometry algorithms for airway length and diameter were successfully verified against previously published manual and automated tracheobronchial airway morphometry data derived from two identical in-situ Balb/c mouse lung casts. There was also excellent agreement in tracheobronchial airway length and diameter between the automated and manual airway data for the AJ, C57BL/6, and Apoe(-/-) mice. Differences in branch angle measurements were partially due to the differences in definition between the automated algorithms and manual morphometry techniques. Unlike the manual airway morphometry techniques, the automated algorithms were able to provide a value for inclination to gravity for each airway. Inclusion of an inclination to gravity angle for each airway along with airway length, diameter, and branch angle make the current automated tracheobronchial airway data suitable for use in dosimetry programs that can provide dosimetry estimates for inhaled material. The significant differences in upper tracheobronchial airways between Balb/c mice and between C57BL/6 and Apoe(-/-) mice highlight the need for mouse strain-specific aerosol dosimetry estimates.

Abstract  The frequent use of rodent hepatic in vitro systems in pharmacological and toxicological investigations challenges extrapolation of in vitro results to the situation in vivo and interspecies extrapolation from rodents to humans. The toxicogenomics approach may aid in evaluating relevance of these model systems for human risk assessment by direct comparison of toxicant-induced gene expression profiles and infers mechanisms between several systems. In the present study, acetaminophen (APAP) was used as a model compound to compare gene expression responses between rat and human using in vitro cellular models, hepatocytes, and between rat in vitro and in vivo. Comparison at the level of modulated biochemical pathways and biological processes rather than at that of individual genes appears preferable as it increases the overlap between various systems. Pathway analysis by T-profiler revealed similar biochemical pathways and biological processes repressed in rat and human hepatocytes in vitro, as well as in rat liver in vitro and in vivo. Repressed pathways comprised energy-consuming biochemical pathways, mitochondrial function, and oxidoreductase activity. The present study is the first that used a toxicogenomics-based parallelogram approach, extrapolating in vitro to in vivo and interspecies, to reveal relevant mechanisms indicative of APAP-induced liver toxicity in humans in vivo.

Abstract  Determining the fate of inhaled aerosols in the respiratory system is essential in assessing the potential toxicity of inhaled airborne materials, responses to airborne pathogens, or in improving inhaled drug delivery. The availability of high-resolution clinical lung imaging and advances in the reconstruction of lung airways from CT images have led to the development of subject-specific in-silico 3D models of aerosol dosimetry, often referred to as computational fluid-particledynamics (CFPD) models. As CFPD models require extensive computing resources, they are typically confined to the upper and large airways. These models can be combined with lowerdimensional models to form multiscale models that predict the transport and deposition of inhaled aerosols in the entire respiratory tract. Understanding where aerosols deposit is only the first of potentially several key events necessary to predict an outcome, being a detrimental health effect or a therapeutic response. To that end, multiscale approaches that combine CFPD with physiologically-based pharmacokinetics (PBPK) models have been developed to evaluate the absorption, distribution, metabolism, and excretion (ADME) of toxic or medicinal chemicals in one or more compartments of the human body. CFPD models can also be combined with host cell dynamics (HCD) models to assess regional immune system responses. This paper reviews the state of the art of these different multiscale approaches and discusses the potential role of personalized or subject-specific modeling in respiratory health.