Effects of Formulation Variables on Lung Dosimetry of Albuterol Sulfate Suspension and Beclomethasone Dipropionate Solution Metered Dose Inhalers.

The performance of pressurized metered dose inhalers (MDIs) is affected by formulation and device variables that impact delivered dose, aerodynamic particle size distribution, and consequently lung deposition and therapeutic effect. Specific formulation variables of relevance to two commercially available products-Proventil® HFA [albuterol sulfate (AS) suspension] and Qvar® [beclomethasone dipropionate (BDP) solution]-were evaluated to determine their influence on key performance attributes measured experimentally with in vitro cascade impaction studies. These commercial MDIs, utilized as model systems, provided mid-points for a design of experiments (DoE) plan to manufacture multiple suspension and solution MDI formulations. The experimental results were utilized as input variables in a computational dosimetry model to predict the effects of MDI formulation variables on lung deposition. For the BDP solution DoE MDIs, increased concentrations of surfactant oleic acid (0-2% w/w) increased lung deposition from 24 to 46%, whereas changes in concentration of the cosolvent ethanol (7-9% w/w) had no effect on lung deposition. For the AS suspension DoE MDIs, changes in oleic acid concentration (0.005-0.25% w/w) did not have significant effects on lung deposition, whereas lung deposition decreased from 48 to 26% as ethanol concentration increased from 2 to 20% w/w, and changes in micronized drug volumetric median particle size distribution (X50, 1.4-2.5 µm) increased deposition in the tracheobronchial airways from 5 to 11%. A direct correlation was observed between fine particle fraction and predicted lung deposition. These results demonstrate the value of using dosimetry models to further explore relationships between performance variables and lung deposition.

Component-specific, cigarette particle deposition modeling in the human respiratory tract.

Inhalation of cigarette smoke particles (CSP) leads to adverse health effects in smokers. Determination of the localized dose to the lung of the inhaled smoke aids in determining vulnerable sites, and identifying components of the smoke that may be responsible for the adverse effects; thus providing a roadmap for harm reduction of cigarette smoking. A particle deposition model specific to CSP was developed for the oral cavity and the lung by accounting for cigarette particle size growth by hygroscopicity, phase change and coagulation. In addition, since the cigarette puff enters the respiratory tract as a dense cloud, the cloud effect on particle drag and deposition was accounted for in the deposition model. Models of particle losses in the oral cavities were developed during puff drawing and subsequent mouth-hold. Cigarette particles were found to grow by hygroscopicity and coagulation, but to shrink as a result of nicotine evaporation. The particle size reached a plateau beyond which any disturbances in the environmental conditions caused the various mechanisms to balance each other out and the particle size remain stable. Predicted particle deposition considering the cloud effects was greater than when treated as a collection of non-interacting particles (i.e. no cloud effects). Accounting for cloud movement provided the necessary physical mechanism to explain the greater than expected, experimentally observed and particle deposition. The deposition model for CSP can provide the necessary input to determine the fate of inhaled CSP in the lung. The knowledge of deposition will be helpful for health assessment and identification and reduction of harmful components of CSP.

Computational fluid dynamics simulations of inhaled nano- and microparticle deposition in the rhesus monkey nasal passages.

Anatomically accurate computational fluid dynamics (CFD) models of the nasal passages of an infant (6 months old, 1.3 kg) and adult (7 years old, 11.9 kg) rhesus monkey were used to predict nasal deposition of inhaled nano- and microparticles. Steady-state, inspiratory airflow simulations were conducted at flow rates equal to 100, 200 and 300% of the estimated minute volume for resting breathing in each model. Particle transport and deposition simulations were conducted using the Lagrangian method to track the motion of inhaled particles. Nasal deposition fractions were higher in the infant model than the adult model at equivalent physiologic flow rates. Deposition curves collapsed when differences in nasal geometry were accounted for by plotting microparticle deposition versus the Stokes number and nanoparticle deposition as a function of the Schmidt number and diffusion parameter. Particle deposition was also quantified on major nasal epithelial types. Maximum olfactory deposition ranged from 5 to 14% for 1-2 nm particles in the adult and infant models, depending on flow rate. For these particle sizes, deposition on respiratory/transitional epithelia ranged from 40 to 50%. Increased deposition was also predicted for olfactory and respiratory/transitional epithelia for particle sizes >5 µm in the infant model and >8 µm in the adult model. Semi-empirical curves were developed based on the CFD simulation results to allow for simplified calculations of age-based deposition in the rhesus monkey nasal passages that can be implemented into lung dosimetry models.

Effect of smoking parameters on the particle size distribution and predicted airway deposition of mainstream cigarette smoke.

It is known that puffing conditions such as puff volume, duration, and frequency vary substantially among individual smokers. This study investigates how these parameters affect the particle size distribution and concentration of fresh mainstream cigarette smoke (MCS) and how these changes affect the predicted deposition of MCS particles in a model human respiratory tract. Measurements of the particle size distribution made with an electrical low pressure impactor for a variety of puffing conditions are presented. The average flow rate of the puff is found to be the major factor effecting the measured particle size distribution of the MCS. The results of these measurements were then used as input to a deterministic dosimetry model (MPPD) to estimate the changes in the respiratory tract deposition fraction of smoke particles. The MPPD dosimetry model was modified by incorporating mechanisms involved in respiratory tract deposition of MCS: hygroscopic growth, coagulation, evaporation of semivolatiles, and mixing of the smoke with inhaled dilution air. The addition of these mechanisms to MPPD resulted in reasonable agreement between predicted airway deposition and human smoke retention measurements. The modified MPPD model predicts a modest 10% drop in the total deposition efficiency in a model human respiratory tract as the puff flow rate is increased from 1050 to 3100 ml/min, for a 2-s puff.

Deposition of ultrafine (nano) particles in the human lung.

Increased production of industrial devices constructed with nanostructured materials raises the possibility of environmental and occupational human exposure with consequent adverse health effects. Ultrafine (nano) particles are suspected of having increased toxicity due to their size characteristics that serve as carrier transports. For this reason, it is critical to refine and improve existing deposition models in the nano-size range. A mathematical model of nanoparticle transport by airflow convection, axial diffusion, and convective mixing (dispersion) was developed in realistic stochastically generated asymmetric human lung geometries. The cross-sectional averaged convective-diffusion equation was solved analytically to find closed-form solutions for particle concentration and losses per lung airway. Airway losses were combined to find lobar, regional, and total lung deposition. Axial transport by diffusion and dispersion was found to have an effect on particle deposition. The primary impact was in the pulmonary region of the lung for particles larger than 10 nm in diameter. Particles below 10 nm in diameter were effectively removed from the inhaled air in the tracheobronchial region with little or no penetration into the pulmonary region. Significant variation in deposition was observed when different asymmetric lung geometries were used. Lobar deposition was found to be highest in the left lower lobe. Good agreement was found between predicted depositions of ultrafine (nano) particles with measurements in the literature. The approach used in the proposed model is recommended for more realistic assessment of regional deposition of diffusion-dominated particles in the lung, as it provides a means to more accurately relate exposure and dose to lung injury and other biological responses.

Quantitative fluorescence of 5-FU-treated fetal rat limbs using confocal laser scanning microscopy and Lysotracker Red.

BACKGROUND: LysoTracker Red (LT) is a paraformaldehyde fixable probe that concentrates into acidic compartments of cells and tissues. After cell death, a high level of lysosomal activity (acidic enzyme) is expressed in tissues resulting from phagocytosis of apoptotic bodies by neighboring cells. LT was shown previously to be an indicator of cell death in a manner similar to other standard assays (Annexin, terminal dUTP nick end labeling, Nile blue sulfate, neutral red, and acridine orange). METHODS: LT fluorescence in fetal rat hindlimbs at gestational day 14 was measured 8 h after administration of the teratogen, 5-fluorouracil (5-FU), with the use of confocal laser scanning microscopy (CLSM). Four dose levels of 5-FU (0, 20, 30, and 40 mg/kg) were studied. The preparation technique involved staining with LT, paraformaldehyde fixation, methanol dehydration, and clearance with benzyl alcohol and benzyl benzoate. After this treatment, the limb was nearly transparent and ready for CLSM analysis. RESULTS: LT staining was observed in specific regions undergoing apoptosis in normal (control) hindlimbs. After 5-FU treatment, highly fluorescent regions appeared in the progress zone (PZ) of the limb. A dose-dependent response to 5-FU treatment was observed. Compared with controls, hindlimbs treated with 20, 30, and 40 mg/kg of 5-FU exhibited more fluorescence within the highly proliferative PZ. These results showed a dose-response relation between 5-FU exposure and LT uptake. CONCLUSIONS: We found that three-dimensional volumetric regions indicating a high level of fluorescence in the embryonic limb bud can be quantified with three different computer analysis programs. The combination of a sample preparation procedure that clears tissue, a CLSM technique that addresses the equipment variables, and an application of statistical population analysis procedures enabled the visualization and quantification of fluorescence in entire fetal rat hindlimbs that were approximately 500 microm in thickness.