The dynamics of SARS-CoV-2 infectivity with changes in aerosol microenvironment.

Understanding the factors that influence the airborne survival of viruses such as severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) in aerosols is important for identifying routes of transmission and the value of various mitigation strategies for preventing transmission. We present measurements of the stability of SARS-CoV-2 in aerosol droplets (∼5 to 10 µm equilibrated radius) over timescales spanning 5 s to 20 min using an instrument to probe survival in a small population of droplets (typically 5 to 10) containing ∼1 virus/droplet. Measurements of airborne infectivity change are coupled with a detailed physicochemical analysis of the airborne droplets containing the virus. A decrease in infectivity to ∼10% of the starting value was observable for SARS-CoV-2 over 20 min, with a large proportion of the loss occurring within the first 5 min after aerosolization. The initial rate of infectivity loss was found to correlate with physical transformation of the equilibrating droplet; salts within the droplets crystallize at relative humidities (RHs) below 50%, leading to a near-instant loss of infectivity in 50 to 60% of the virus. However, at 90% RH, the droplet remains homogenous and aqueous, and the viral stability is sustained for the first 2 min, beyond which it decays to only 10% remaining infectious after 10 min. The loss of infectivity at high RH is consistent with an elevation in the pH of the droplets, caused by volatilization of CO2 from bicarbonate buffer within the droplet. Four different variants of SARS-CoV-2 were compared and found to have a similar degree of airborne stability at both high and low RH.

An assessment of the airborne longevity of group A Streptococcus.

Group A streptococcus (GAS) infections result in more than 500 000 deaths annually. Despite mounting evidence for airborne transmission of GAS, little is known about its stability in aerosol. Measurements of GAS airborne stability were carried out using the Controlled Electrodynamic Levitation and Extraction of Bioaerosols onto a Substrate (CELEBS) instrument. CELEBS measurements with two different isolates of GAS suggest that it is aerostable, with approximately 70 % of bacteria remaining viable after 20 min of levitation at 50 % relative humidity (RH), with lower survival as RH was reduced. GAS airborne viability loss was driven primarily by desiccation and efflorescence (i.e. salt crystallization), with high pH also potentially playing a role, given reduced survival in bicarbonate containing droplet compositions. At low enough RH for efflorescence to occur, a greater proportion of organic components in the droplet appeared to protect the bacteria from efflorescence. These first insights into the aerosol stability of GAS indicate that airborne transmission of these respiratory tract bacteria may occur, and that both the composition of the droplet containing the bacteria, and the RH of the air affect the duration of bacterial survival in this environment. Future studies will explore a broader range of droplet and air compositions and include a larger selection of GAS strains.

Toward Standardized Aerovirology: A Critical Review of Existing Results and Methodologies.

Understanding the airborne survival of viruses is important for public health and epidemiological modeling and potentially to develop mitigation strategies to minimize the transmission of airborne pathogens. Laboratory experiments typically involve investigating the effects of environmental parameters on the viability or infectivity of a target airborne virus. However, conflicting results among studies are common. Herein, the results of 34 aerovirology studies were compared to identify links between environmental and compositional effects on the viability of airborne viruses. While the specific experimental apparatus was not a factor in variability between reported results, it was determined that the experimental procedure was a major factor that contributed to discrepancies in results. The most significant contributor to variability between studies was poorly defined initial viable virus concentration in the aerosol phase, causing many studies to not measure the rapid inactivation, which occurs quickly after particle generation, leading to conflicting results. Consistently, studies that measured their reference airborne viability minutes after aerosolization reported higher viability at subsequent times, which indicates that there is an initial loss of viability which is not captured in these studies. The composition of the particles which carry the viruses was also found to be important in the viability of airborne viruses; however, the mechanisms for this effect are unknown. Temperature was found to be important for aerosol-phase viability, but there is a lack of experiments that directly compare the effects of temperature in the aerosol phase and the bulk phase. There is a need for repeated measurements between different research groups under identical conditions both to assess the degree of variability between studies and also to attempt to better understand already published data. Lack of experimental standardization has hindered the ability to quantify the differences between studies, for which we provide recommendations for future studies. These recommendations are as follows: measuring the reference airborne viability using the "direct method"; use equipment which maximizes time resolution; quantify all losses appropriately; perform, at least, a 5- and 10-min sample, if possible; report clearly the composition of the virus suspension; measure the composition of the gas throughout the experiment. Implementing these recommendations will address the most significant oversights in the existing literature and produce data which can more easily be quantitatively compared.

Studies of the Crystallization and Dissolution of Individual Suspended Sodium Chloride Aerosol Particles.

Aerosols transform between physical phases, as they respond to variations in environmental conditions. There are many industries that depend on these dynamic processes of crystallization and dissolution. Here, a single particle technique (an electrodynamic balance) is used to explore the crystallization and dissolution dynamics of a model system, sodium chloride. The physical and environmental factors that influence the dynamics of crystal formation from a saline droplet (whose initial radius is ∼25 µm) and the kinetics of water adsorption onto dried particles are examined. The drying relative humidity (RH) is shown to impact the physical properties of the dried particle. When a saline droplet is injected into an airflow at an RH close to the efflorescence RH (ERH, 45%), an individual single crystal forms. By contrast, when a compositionally equivalent saline droplet is injected into dry air (RH ∼ 0%), a salt crystal made of multiple crystalline particles is formed. Subsequent to crystallization, the crystal shape, morphology, and surface area were all found to affect the dissolution dynamics of the dried particle. Additionally, we report that the difference between the deliquesce RH and environmental RH significantly impacts the dissolution time scale.

Optical Interrogation of Single Levitated Droplets in a Linear Quadrupole Trap by Cavity Ring-Down Spectroscopy.

Optical trapping is a well-established technique to manipulate and levitate micro- and nanoscale particles and droplets. However, optical traps for single aerosol studies are most often limited to trapping spherical nonabsorbing droplets, and a universal optical trap for the stable confinement of particles regardless of their absorption strength and morphology is not established. Instead, new opportunities arise from levitating droplets using electrodynamic traps. Here, using a combined electrodynamic linear quadrupole trap and a cavity ring-down spectrometer, we demonstrate that it is possible to trap single droplets and simultaneously measure their extinction cross sections and elastic scattering phase functions over extended periods of time. To test the novel setup, we evaluated the evaporation of 1,2,6-hexanetriol under low-humidity conditions, and the evolution of aqueous (NH4)2SO4 and NaCl droplets experiencing changing environmental conditions. Our studies extended beyond spherical droplets and we measured particle extinction cross sections after the efflorescence (crystallization) of the inorganic salt particles. Comparison of measured cross sections for crystallized particles with light scattering model predictions (using Mie theory or the T-matrix/extended boundary-condition method (EBCM) implementations for random orientation, with either the spheroid or superellipsoid parameterizations) enables information on particle shape to be inferred. Specifically, we find that cross sections for dry (NH4)2SO4 particles are accounted for by Mie theory and, thus, particle shape is represented well by a sphere. Conversely, the cross sections for dry NaCl particles are only reconciled with light scattering models pertaining to nonspherical shapes. These results will have implications for accurate remote sensing retrievals of dry salt optical properties and for parameterizations implemented in radiative forcing calculations with changing humidity. Moreover, our new platform for precise and accurate measurement of optical properties of micron-scale and sub-micron particles has potential applications in a range of areas of atmospheric science, such as precise light scattering measurements for ice crystals and mineral dust. It represents a promising step toward accurate characterizations of optical properties for nonspherical and light-absorbing aerosols.

Transformative Approach To Investigate the Microphysical Factors Influencing Airborne Transmission of Pathogens.

Emerging outbreaks of airborne pathogenic infections worldwide, such as the current severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) pandemic, have raised the need to understand parameters affecting the airborne survival of microbes in order to develop measures for effective infection control. We report a novel experimental strategy, TAMBAS (tandem approach for microphysical and biological assessment of airborne microorganism survival), to explore the synergistic interactions between the physicochemical and biological processes that impact airborne microbe survival in aerosol droplets. This innovative approach provides a unique and detailed understanding of the processes taking place from aerosol droplet generation through to equilibration and viability decay in the local environment, elucidating decay mechanisms not previously described. The impact of evaporation kinetics, solute hygroscopicity and concentration, particle morphology, and equilibrium particle size on airborne survival are reported, using Escherichia coli MRE162 as a benchmark system. For this system, we report that (i) particle crystallization does not directly impact microbe longevity, (ii) bacteria act as crystallization nuclei during droplet drying and equilibration, and (iii) the kinetics of size and compositional change appear to have a larger effect on microbe longevity than the equilibrium solute concentration.IMPORTANCE A transformative approach to identify the physicochemical processes that impact the biological decay rates of bacteria in aerosol droplets is described. It is shown that the evaporation process and changes in the phase and morphology of the aerosol particle during evaporation impact microorganism viability. The equilibrium droplet size was found to affect airborne bacterial viability. Furthermore, the presence of Escherichia coli MRE162 in a droplet does not affect aerosol growth/evaporation but influences the dynamic behavior of the aerosol by processing the culture medium prior to aerosolization, affecting the hygroscopicity of the culture medium; this highlights the importance of the inorganic and organic chemical composition within the aerosolized droplets that impact hygroscopicity. Bacteria also act as crystallization nuclei. The novel approach and data have implications for increased mechanistic understanding of aerosol survival and infectivity in bioaerosol studies spanning the medical, veterinary, farming, and agricultural fields, including the role of microorganisms in atmospheric processing and cloud formation.

Coalescence Sampling and Analysis of Aerosols using Aerosol Optical Tweezers.

We present a first exploratory study to assess the use of aerosol optical tweezers as an instrument for sampling and detecting accumulation- and coarse-mode aerosol. A subpicoliter aqueous aerosol droplet is captured in the optical trap and used as a sampling volume, accreting mass from a free-flowing aerosol generated by a medical nebulizer or atomizer. Real-time measurements of the initial stability in size, refractive index, and composition of the sampling droplet inferred from Raman spectroscopy confirm that these quantities can be measured with high accuracy and low noise. Typical standard deviations in size and refractive index of the sampling droplet over a period of 200 s are <±2 nm and <±0.0005, respectively, equivalent to <±0.04% in both measured quantities. A standard deviation of <±1% over a 200 s period is achieved in the spontaneous Raman intensity measurement. When sampling coarse-mode aerosol, mass changes of <10 pg can be detected by the sampling droplet as discrete coalescence events. With accumulation-mode aerosol, we show that fluxes as low as 0.068 pg s-1 can be detected over a 50 s period, equivalent to ∼3 pg of sampled material.

Aerobiology: Experimental Considerations, Observations, and Future Tools.

Understanding airborne survival and decay of microorganisms is important for a range of public health and biodefense applications, including epidemiological and risk analysis modeling. Techniques for experimental aerosol generation, retention in the aerosol phase, and sampling require careful consideration and understanding so that they are representative of the conditions the bioaerosol would experience in the environment. This review explores the current understanding of atmospheric transport in relation to advances and limitations of aerosol generation, maintenance in the aerosol phase, and sampling techniques. Potential tools for the future are examined at the interface between atmospheric chemistry, aerosol physics, and molecular microbiology where the heterogeneity and variability of aerosols can be explored at the single-droplet and single-microorganism levels within a bioaerosol. The review highlights the importance of method comparison and validation in bioaerosol research and the benefits that the application of novel techniques could bring to increasing the understanding of aerobiological phenomena in diverse research fields, particularly during the progression of atmospheric transport, where complex interdependent physicochemical and biological processes occur within bioaerosol particles.

Measurements and Predictions of Binary Component Aerosol Particle Viscosity.

Organic aerosol particles are known to often absorb/desorb water continuously with change in gas phase relative humidity (RH) without crystallization. Indeed, the prevalence of metastable ultraviscous liquid or amorphous phases in aerosol is well-established with solutes often far exceeding bulk phase solubility limits. Particles are expected to become increasingly viscous with drying, a consequence of the plasticizing effect of water. We report here measurements of the variation in aerosol particle viscosity with RH (equal to condensed phase water activity) for a range of organic solutes including alcohols (diols to hexols), saccharides (mono-, di-, and tri-), and carboxylic acids (di-, tri-, and mixtures). Particle viscosities are measured over a wide range (10-3 to 1010 Pa s) using aerosol optical tweezers, inferring the viscosity from the time scale for a composite particle to relax to a perfect sphere following the coalescence of two particles. Aerosol measurements compare well with bulk phase studies (well-within an order of magnitude deviation at worst) over ranges of water activity accessible to both. Predictions of pure component viscosity from group contribution approaches combined with either nonideal or ideal mixing reproduce the RH-dependent trends particularly well for the alcohol, di-, and tricarboxylic acid systems extending up to viscosities of 104 Pa s. By contrast, predictions overestimate the viscosity by many orders of magnitude for the mono-, di-, and trisaccharide systems, components for which the pure component subcooled melt viscosities are ≫1012 Pa s. When combined with a typical scheme for simulating the oxidation of α-pinene, a typical atmospheric pathway to secondary organic aerosol (SOA), these predictive tools suggest that the pure component viscosities are less than 106 Pa s for ∼97% of the 50,000 chemical products included in the scheme. These component viscosities are consistent with the conclusion that the viscosity of α-pinene SOA is most likely in the range 105 to 108 Pa s. Potential improvements to the group contribution predictive tools for pure component viscosities are considered.

Influence of organic films on the evaporation and condensation of water in aerosol.

Uncertainties in quantifying the kinetics of evaporation and condensation of water from atmospheric aerosol are a significant contributor to the uncertainty in predicting cloud droplet number and the indirect effect of aerosols on climate. The influence of aerosol particle surface composition, particularly the impact of surface active organic films, on the condensation and evaporation coefficients remains ambiguous. Here, we report measurements of the influence of organic films on the evaporation and condensation of water from aerosol particles. Significant reductions in the evaporation coefficient are shown to result when condensed films are formed by monolayers of long-chain alcohols [C(n)H(2n+1)OH], with the value decreasing from 2.4 × 10(-3) to 1.7 × 10(-5) as n increases from 12 to 17. Temperature-dependent measurements confirm that a condensed film of long-range order must be formed to suppress the evaporation coefficient below 0.05. The condensation of water on a droplet coated in a condensed film is shown to be fast, with strong coherence of the long-chain alcohol molecules leading to islanding as the water droplet grows, opening up broad areas of uncoated surface on which water can condense rapidly. We conclude that multicomponent composition of organic films on the surface of atmospheric aerosol particles is likely to preclude the formation of condensed films and that the kinetics of water condensation during the activation of aerosol to form cloud droplets is likely to remain rapid.

Dynamics of Particle Size on Inhalation of Environmental Aerosol and Impact on Deposition Fraction.

Inhalation of elevated levels of particulate air pollution has been shown to elicit the onset of adverse health effects in humans, where the magnitude of the response is a product of where in the lung the particulate dose is delivered. At any point in time during inhalation the depositional flux of the aerosol is a function of the radius of the droplet, thus a detailed understanding of the rate and magnitude of the mass flux of water to the droplet during inhalation is crucial. In this study, we assess the impact of aerosol hygroscopicity on deposited dose through the inclusion of a detailed treatment of the mass flux of water to account for the dynamics of particle size in a modified version of the standard International Commission on Radiological Protection (ICRP) whole lung deposition model. The ability to account for the role of the relative humidity (RH) of the aerosol prior to, and during, inhalation on the deposition pattern is explored, and found to have a significant effect on the deposition pattern. The model is verified by comparison to previously published measurements, and used to demonstrate that ambient RH affects where in the lung indoor particulate air pollution is delivered.

Simultaneous analysis of the equilibrium hygroscopicity and water transport kinetics of liquid aerosol.

We demonstrate that the equilibrium hygroscopic response of an aerosol droplet and the kinetics of water condensation and evaporation can be retrieved with high accuracy, even close to saturation, through comparative measurements of probe and sample aerosol droplets. The experimental methodology is described and is based on an electrodynamic balance with a newly designed trapping chamber. Through use of a probe aerosol, composed of either pure water or a sodium chloride solution of known concentration, the gas-phase relative humidity (RH) can be accurately measured with an uncertainty of typically <0.005. By fast manipulation of the airflows into the chamber, a step-change in RH over a time scale of <0.5 s can be achieved. Using this approach, the kinetics of mass transfer are studied using the comparative procedure, and results are compared to theoretical mass flux predictions. The time-dependent measured mass fluxes for sodium chloride, ammonium sulfate, sorbitol, and galactose are used to calculate droplet water activities as a function of the droplet growth factor, allowing retrieval of a hygroscopic growth curve in a matter of seconds. Comparisons with both new and established thermodynamic predictions of hygroscopicity, as well as to optical tweezers measurements, are presented, demonstrating good agreement within the experimental uncertainties.

Formation of a stable mimic of ambient particulate matter containing viable infectious respiratory syncytial virus and its dry-deposition directly onto cell cultures.

Epidemiological associations of worse respiratory outcomes from combined exposure to ambient particulate matter (PM) and respiratory viral infection suggest possible interactions between PM and viruses. To characterize outcomes of such exposures, we developed an in vitro mimic of the in vivo event of exposure to PM contaminated with respiratory syncytial virus (RSV). Concentration of infectious RSV stocks and a particle levitation apparatus were the foundations of the methodology developed to generate specific numbers of PM mimics (PM(Mimics)) of known composition for dry, direct deposition onto airway epithelial cell cultures. Three types of PM(Mimics) were generated for this study: (i) carbon alone (P(C)), (ii) carbon and infectious RSV (P(C+RSV)), and (iii) aerosols consisting of RSV (A(RSV)). P(C+RSV) were stable in solution and harbored infectious RSV for up to 6 months. Unlike A(RSV) infection, P(C+RSV) infection was found to be dynamin dependent and to cause lysosomal rupture. Cells dosed with PM(Mimics) comprised of RSV (A(RSV)), carbon (P(C)), or RSV and carbon (P(C+RSV)) responded differentially as exemplified by the secretion patterns of IL-6 and IL-8. Upon infection, and prior to lung cell death due to viral infection, regression analysis of these two mediators in response to incubation with A(RSV), P(C), or P(C+RSV) yielded higher concentrations upon infection with the latter and at earlier time points than the other PM(Mimics). In conclusion, this experimental platform provides an approach to study the combined effects of PM-viral interactions and airway epithelial exposures in the pathogenesis of respiratory diseases involving inhalation of environmental agents.

Oxidative Stress Contributes to Bacterial Airborne Loss of Viability.

While the airborne decay of bacterial viability has been observed for decades, an understanding of the mechanisms driving the decay has remained elusive. The airborne transport of bacteria is often a key step in their life cycle and as such, characterizing the mechanisms driving the airborne decay of bacteria is an essential step toward a more complete understanding of microbial ecology. Using the Controlled Electrodynamic Levitation and Extraction of Bioaerosols onto a Substrate (CELEBS), it was possible to systematically evaluate the impact of different physicochemical and environmental parameters on the survival of Escherichia coli in airborne droplets of Luria Bertani broth. Rather than osmotic stress driving the viability loss, as was initially considered, oxidative stress was found to play a key role. As the droplets evaporate and equilibrate with the surrounding environment, the surface-to-volume ratio increases, which in turn increased the formation of reactive oxygen species in the droplet. These reactive oxygen species appear to play a key role in driving the airborne loss of viability of E. coli. IMPORTANCE The airborne transport of bacteria has a wide range of impacts, from disease transmission to cloud formation. By understanding the factors that influence the airborne stability of bacteria, we can better understand these processes. However, while we have known for several decades that airborne bacteria undergo a gradual loss of viability, we have not previously identified the mechanisms driving this process. In this work, we discovered that oxygen surrounding an airborne droplet facilitates the formation of reactive oxygen species within the droplet, which then gradually damage and kill bacteria within the droplet. This discovery indicates that adaptations to help bacteria deal with oxidative stress may also aid their airborne survival and be essential adaptations for bacterial airborne pathogens. Understanding the adaptations bacteria need to survive in airborne droplets could eventually lead to the development of novel antimicrobials designed to inhibit their airborne survival, helping to prevent the transmission of disease.

Accounting for changes in particle charge, dry mass and composition occurring during studies of single levitated particles.

The most used instrument in single particle hygroscopic analysis over the past thirty years has been the electrodynamic balance (EDB). Two general assumptions are made in hygroscopic studies involving the EDB. First, it is assumed that the net charge on the droplet is invariant over the time scale required to record a hygroscopic growth cycle. Second, it is assumed that the composition of the droplet is constant (aside from the addition and removal of water). In this study, we demonstrate that these assumptions cannot always be made and may indeed prove incorrect. The presence of net charge in the humidified vapor phase reduces the total net charge retained by the droplet over prolonged levitation periods. The gradual reduction in charge limits the reproducibility of hygroscopicity measurements made on repeated RH cycles with a single particle, or prolonged experiments in which the particle is held at a high relative humidity. Further, two contrasting examples of the influence of changes in chemical composition changes are reported. In the first, simple acid-base chemistry in the droplet leads to the irreversible removal of gaseous ammonia from a droplet containing an ammonium salt on a time scale that is shorter than the hygroscopicity measurement. In the second example, the net charge on the droplet (<100 fC) is high enough to drive redox chemistry within the droplet. This is demonstrated by the reduction of iodic acid in a droplet made solely of iodic acid and water to form iodine and an iodate salt.

Bulk, surface, and gas-phase limited water transport in aerosol.

The influence of solute species on mass transfer to and from aqueous aerosol droplets is investigated using an electrodynamic balance coupled with light scattering techniques. In particular, we explore the limitations imposed on water evaporation by slow bulk phase diffusion and by the formation of surface organic films. Measurements of evaporation from ionic salt solutions, specifically sodium chloride and ammonium sulfate, are compared with predictions from an analytical model framework, highlighting the uncertainties associated with quantifying gas diffusional transport. The influence of low solubility organic acids on mass transfer is reported and compared to both model predictions and previous work. The limiting value of the evaporation coefficient that can be resolved by this approach, when uncertainties in key thermophysical quantities are accounted for, is estimated. The limitation of slow bulk phase diffusion on the evaporation rate is investigated for gel and glass states formed during the evaporation of magnesium sulfate and sucrose droplets, respectively. Finally, the effect of surfactants on evaporation has been probed, with soluble surfactants (such as sodium dodecyl sulfate) leading to little or no retardation of evaporation through slowing of surface layer kinetics.

Mucin Transiently Sustains Coronavirus Infectivity through Heterogenous Changes in Phase Morphology of Evaporating Aerosol.

Respiratory pathogens can be spread though the transmission of aerosolised expiratory secretions in the form of droplets or particulates. Understanding the fundamental aerosol parameters that govern how such pathogens survive whilst airborne is essential to understanding and developing methods of restricting their dissemination. Pathogen viability measurements made using Controlled Electrodynamic Levitation and Extraction of Bioaerosol onto Substrate (CELEBS) in tandem with a comparative kinetics electrodynamic balance (CKEDB) measurements allow for a direct comparison between viral viability and evaporation kinetics of the aerosol with a time resolution of seconds. Here, we report the airborne survival of mouse hepatitis virus (MHV) and determine a comparable loss of infectivity in the aerosol phase to our previous observations of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Through the addition of clinically relevant concentrations of mucin to the bioaerosol, there is a transient mitigation of the loss of viral infectivity at 40% RH. Increased concentrations of mucin promoted heterogenous phase change during aerosol evaporation, characterised as the formation of inclusions within the host droplet. This research demonstrates the role of mucus in the aerosol phase and its influence on short-term airborne viral stability.

Water Uptake by Evaporating pMDI Aerosol Prior to Inhalation Affects Both Regional and Total Deposition in the Respiratory System.

As pulmonary drug deposition is a function of aerosol particle size distribution, it is critical that the dynamics of particle formation and maturation in pMDI sprays in the interim between generation and inhalation are fully understood. This paper presents an approach to measure the evaporative and condensational fluxes of volatile components and water from and to solution pMDI droplets following generation using a novel technique referred to as the Single Particle Electrodynamic Lung (SPEL). In doing so, evaporating aerosol droplets are shown capable of acting as condensation nuclei for water. Indeed, we show that the rapid vaporisation of volatile components from a volatile droplet is directly correlated to the volume of water taken up by condensation. Furthermore, a significant volume of water is shown to condense on droplets of a model pMDI formulation (hydrofluoroalkane (HFA), ethanol and glycerol) during evaporative droplet ageing, displaying a dramatic shift from a core composition of a volatile species to that of predominantly water (non-volatile glycerol remained in this case). This yields a droplet with a water activity of 0.98 at the instance of inhalation. The implications of these results on regional and total pulmonary drug deposition are explored using the International Commission of Radiological Protection (ICRP) deposition model, with an integrated semi-analytical treatment of hygroscopic growth. Through this, droplets with water activity of 0.98 upon inhalation are shown to produce markedly different dose deposition profiles to those with lower water activities at the point of inspiration.

Pulmonary aerosol delivery and the importance of growth dynamics.

Aerosols are dynamic systems, responding to variations in the surrounding environmental conditions by changing in size, composition and phase. Although, widely used in inhalation therapies, details of the processes occurring on aerosol generation and during inhalation have received little attention. Instead, research has focused on improvements to the formulation of the drug prior to aerosolization and the resulting clinical efficacy of the treatment. Here, we highlight the processes that occur during aerosol generation and inhalation, affecting aerosol disposition when deposited and, potentially, impacting total and regional doses. In particular, we examine the response of aerosol particles to the humid environment of the respiratory tract, considering both the capacity of particles to grow by absorbing moisture and the timescale for condensation to occur. [Formula: see text].