Drug targeting to lungs by way of microspheres

In many conventional drug delivery systems in vogue, failure to deliver efficient drug delivery at the target site/organs; is evident as a result, less efficacious pharmacological response is elicited. Microspheres can be derived a remedial measure which can improve site-specific drug delivery to a considerable extent. As an application, Lung-targeting Ofloxacin-loaded gelatin microspheres (GLOME) were prepared by water in oil emulsion method. The Central Composite Design (CCD) was used to optimize the process of preparation, the appearance and size distribution were examined by scanning electron microscopy, the aspects such as in vitro release characteristics, stability, drug loading, loading efficiency, pharmacokinetics and tissue distribution in albino mice were studied. The experimental results showed that the microspheres in the range of 0.32-22 microm. The drug loading and loading efficiency were 61.05 and 91.55% respectively. The in vitro release profile of the microspheres matched the korsmeyer's peppas release pattern, and release at 1h was 42%, while for the original drug, ofloxacin under the same conditions 90.02% released in the first half an hour. After i.v. administration (15 min), the drug concentration of microspheres group in lung in albino mice was 1048 microg/g, while that of controlled group was 6.77 microg/g. GLOME found to release the drug to a maximum extent in the target tissue, lungs.     

Respiratory deposition and inhalability of monodisperse aerosols in Long-Evans rats.

Because of limitations on conducting exposure experiments using human subjects to evaluate adverse health effects, the deposition and fate of airborne particles in animals are often studied. The results of such studies are extrapolated to humans to estimate equivalent dose and subsequent response. In this article, particle inhalability and respiratory deposition of micron-size particles are determined for female Long-Evans rats. Monodisperse aerosols were generated from a solution of radiolabeled iron chloride ((59)FeCl(3)). Long-Evans rats were exposed to the radiolabeled particles in a Cannon nose-only exposure tower to determine head, lung lobar, and total lung deposition fractions. Particle deposition fractions in a hypothetical situation, when all particles are inhalable, were found from an experimentally validated deposition model. Particle inhalability in a Cannon nose-only exposure scenario was obtained by comparing the measured deposition fractions with the predicted values for the case of 100% inhalability. Particle deposition fraction and inhalability were compared with data available in the literature. For large particles, the measured deposition fraction was lower than the literature values. Consequently, our inhalability estimates were found to be lower than previously published values. The findings here will directly affect health risk assessments in humans from exposure to airborne particles. The deposition results will improve the database on particle deposition in the lung airways of rats, and inhalability information will improve the accuracy of rat-to-human data extrapolation.     

A system for the production and delivery of monodisperse salbutamol aerosols to the lungs

An aerosol system is described for the generation and delivery of measured doses of monodisperse therapeutic drug particles to the human lungs. The system comprises a spinning top aerosol generator (STAG), aerosol chamber and inhalation control unit. Monodisperse aerosols allow drug particle size effects to be studied as the dose is within a narrow size distribution and when combined with controlled inhalation may lead to more precise targeting of therapeutic drug to the airways. Using the STAG, particles in the size range 1.5-12 microm were generated and their mass median aerodynamic diameter (MMAD) and concentration measured using an aerodynamic particle sizer (APS). The application and validation of the system with the bronchodilator drug salbutamol sulphate is described, and its potential use in the study of aerosol particle size effects is discussed.     

Production of monodisperse silica gel microspheres for bioencapsulation by extrusion into an oil cross-flow

Silica gel microspheres are ideal materials for bioencapsulation due to their mechanical properties, biocompatibility, and stability. Encapsulated cells are isolated from the environment and protected from predators, changes in pH, and osmotic stress. However methods for the production of silica gel microspheres suitable for bioencapsulation are not well established. This paper describes a method for the production of monodisperse silicon alkoxide cross-linked silica nanoparticle (SNP) gel microspheres for bioencapsulation in which silica gel precursor is extruded from a needle into a cross-flowing stream of mineral oil. Microspheres produced ranged from 1.3 to 2.9?mm in diameter with coefficients of variation ranging from 2 to 6%. Microsphere size was mainly controlled by the flowrate of the cross-flowing oil and smaller microspheres generally had larger coefficients of variation. The method described in this paper can be optimised to produce silica gel microspheres with a diverse range of compositions and properties.     

Particle deposition in children's lungs: theory and experiment.

A mathematical model of inhaled aerosol particle deposition for children is presented and validated with data from two published experimental studies. The model accurately predicts deposition fraction (DF) in children as a function of particle size for particles in the size range 1-3 microns for both sedentary and exercise breathing conditions. When the experimental data are grouped according to age, the model is able to predict age-dependent trends in DF at the studied particle sizes under sedentary breathing conditions. The model predicts that when ventilatory conditions are held constant, age-dependent changes in morphology result in decreasing DF with age; however, under realistic conditions these changes may be masked by age-dependent changes in ventilation. Despite the fact that mean DF differs significantly from adult values only in children younger than 9, the model predicted that dose-per-surface area may still be greater in children due to smaller lung sizes.     

Ozone-induced alterations in collagen metabolism of rat lungs

Rats were exposed to amounts of ozone ranging from 0.5 to 2.0 ppm for intervals of 1, 2, or 3 weeks. Collagen synthesis rates in their lungs were quantitated by biochemical analyses performed with lung minces. Correlative histological observations were made in different lung lobes from the same rats. At all levels of ozone tested, collagen synthesis rates of the lungs were significantly elevated and histologically discernible fibrosis of the alveolar duct walls was observed. Within the range of ozone concentrations studied, the elevation of collagen synthesis rate in exposed rats was a linear function of the level of ozone to which the animals were exposed. We conclude that exposure of rats to near-ambient levels of ozone causes biochemically and histologically discernible fibrotic changes in their lungs, suggesting that such effects may occur at levels of ozone at or near the current ambient air quality standard for this pollutant.     

Macromolecule release from monodisperse PLG microspheres: control of release rates and investigation of release mechanism

Novel macromolecular therapeutics such as peptides, proteins, and DNA are advancing rapidly toward the clinic. Because of typically low oral bioavailability, macromolecule delivery requires invasive methods such as frequently repeated injections. Parenteral depots including biodegradable polymer microspheres offer the possibility of reduced dosing frequency but are limited by the inability to adequately control delivery rates. To control release and investigate release mechanisms, we have encapsulated model macromolecules in monodisperse poly(D,L-lactide-co-glycolide) (PLG) microspheres using a double-emulsion method in combination with the precision particle fabrication technique. We encapsulated fluorescein-dextran (F-Dex) and sulforhodamine B-labeled bovine serum albumin (R-BSA) into PLG microspheres of three different sizes: 31, 44, and 80 microm and 34, 47, and 85 microm diameter for F-Dex and R-BSA, respectively. The in vitro release profiles of both compounds showed negligible initial burst. During degradation and release, the microspheres hollowed and swelled at critical time points dependant upon microsphere size. The rate of these events increased with microsphere size resulting in the largest microspheres exhibiting the fastest overall release rate. Monodisperse microspheres may represent a new delivery system for therapeutic proteins and DNA and provide enhanced control of delivery rates using simple injectable depot formulations.     

Effect of inhaled terbutaline sulphate in relation to its deposition in the lungs

We studied the effects of inhaled terbutaline on FEV₁ and gas exchange, and the pattern of deposition within the lungs. To document this and to estimate the dose of terbutaline administered to the lungs, [^99mTc]DTPA was added to nebulised terbutaline solution. The aerosol was deposited preferentially in large or small airways by using aerosols with different particle mass median diameters (1.5 and 4.8 μm) and different inhalation flow rates (0.25 and 1.01/s). The patients inhaled placebo and then three increasing doses of terbutaline (0.006, 0.02 and 0.08 mg to the lungs). Finally, 2 mg terbutaline was inhaled from a metered dose inhaler via a spacer. After each inhalation FEV₁, PaO₂ and PaCO₂ was measured. The inhalation of small particles at a low flow resulted in a fairly uniform lung deposition, while larger particles at a higher flow resulted in heavy central deposition. Penetration index for small and large particles were 1.3 ± 0.2 and 0.8 ± 0.3 (P < 0.001), respectively. In both groups FEV₁ increased similarly with each dose, and at 0.02 and 0.08 mg the increase was significant (P < 0.01). After eight metered doses of terbutaline sulphate (0.25 mg per dose) inhaled via a spacer, there was a further increase in FEV₁ (P < 0.001). Gas exchange did not differ between the two groups but if they were combined the Da-aO₂ was significantly lower after metered doses than control (P < 0.05). Thus, it appears that the site of deposition is not important for the bronchodilator effect of terbutaline, and gas exchange tended to improve with both modes of administration.     

Polyamine synthesis in rat lungs injured with alpha-naphthylthiourea

The diamine, putrescine, and polyamines, spermidine and spermine, are low molecular weight organic cations with documented regulatory roles in cell growth and differentiation. Multiple lines of direct and indirect evidence suggest that these organic cations also may function in stimulus-response coupling processes regulating cellular injury and repair. For example, recent studies in monocrotaline-treated rats, hyperoxic rats, and in cultured pulmonary endothelial cells suggest that polyamines regulate pulmonary endothelial integrity and may thus participate in development and/or regression of acute edematous lung injury. To determine if the polyamines are involved in a well-characterized animal model of acute lung injury, the present experiments assessed the relation between changes in polyamine synthesis and development of edema in lungs from rats treated with alpha-naphthylthiourea (ANTU). ANTU caused dose- and time-dependent increases in the lung activity of the initial and rate-limiting enzyme in polyamine biosynthesis, ornithine decarboxylase (ODC) and in the lung contents of the polyamines putrescine, spermidine, and spermine. ANTU also caused dose- and time-dependent increases in the lung wet-to-dry weight ratio indicative of pulmonary edema formation. Changes in lung polyamine biosyntheic activity after ANTU did not relate temporally to changes in the lung wet-to-dry weight ratio: ODC activity was depressed during the 3-h period immediately following ANTU administration, a period when the wet-to-dry weight ratio was increasing, and markedly elevated at 18 h after ANTU administration when the wet-to-dry weight ratio had returned to control levels. Pretreatment of the animals with alpha-difluoromethylornithine, a highly specific inhibitor of ODC, failed to attenuate ANTU-induced increases in lung wet-to-dry weight ratio. These observations indicate polyamine synthesis is enhanced in rat lungs with ANTU-induced pulmonary edema but, unlike certain other models of lung injury and pulmonary edema, accumulation of polyamines probably is not essential for development of edematous lung injury. It is conceivable that in this animal model polyamines play a role in lung repair processes or some longer-term consequence of lung injury.     

In vitro monodisperse aerosol deposition in a mouth and throat with six different inhalation devices.

Experiments were performed to determine the effect of different pharmaceutical aerosol inhalation devices on the deposition of monodisperse aerosols in an idealized mouth and throat geometry. The devices included two dry powder inhalers (Diskus and Turbuhaler), two nebulizers (Pari LC STAR and Hudson T-Updraft), and a metered dose inhaler with attached holding chamber (Aerochamber), in addition to a straight tube (1.7 cm inner diameter). Aerosol particles (DL-alpha tocopheryl acetate) of diameters of 2.5, 5, and 7 microm generated by a vibrating orifice generator were inhaled at steady air flow rates of Q = 5-90 L/min through the devices and into the mouth-throat. Deposition in the mouth-throat and after-filter were determined by ultraviolet (UV) spectrophotometric assay. The amount of deposition in the mouth and throat region was found to depend on the type of device that the aerosol entered through. Deposition in the extrathoracic region with the two types of jet nebulizers did not differ significantly (p > 0.1) from that of a straight tube or each other over their entire tested range of 590 > or = pd2Q > or = 11,375, where p is particle density (in g/cm3), d is particle diameter (in microm), and Q is flow rate (in cm3/s). The metered dose inhaler with attached holding chamber was found to differ from the straight tube only at two intermediate values of pd2Q = 5,145 and 16,033. The deposition occurring for the dry powder inhalers was found to be significantly greater than for the straight tube for all values of pd2Q > or = 10,954 for the Diskus and pd2Q > or = 9,435 for the Turbuhaler. Deposition with the dry powder inhalers was found to be up to 14 times greater than that with the straight tube. Thus, the inhaler geometry that the aerosol passes through prior to entering the mouth and throat region can greatly affect the deposition in the mouth-throat.     

Computer simulations of particle deposition in the lungs of chronic obstructive pulmonary disease patients

Epidemiology data show that mortality rates for chronic obstructive pulmonary disease (COPD) patients increase with an increase in concentration of ambient particulate matter (PM). This is not seen for normal subjects. Therefore, the U.S. Environmental Protection Agency (EPA) has identified COPD patients as a susceptible subpopulation to be considered in regulatory standards. In the present study, a computer model was used to calculate deposition fractions of PM within the lungs of COPD patients. The morphology of COPD lungs was characterized by two distinct components: obstruction of airways (chronic bronchitis component), and degeneration of alveolar structure (emphysema component). The chronic bronchitis component was modeled by reducing airway diameters using airway resistance measurements in vivo, and the emphysema component was modeled by increasing alveolar volumes. Calculated results were compared with experimental data obtained from COPD patients for controlled breathing trials (tidal volume of 500 ml, respiratory time of 1 s) with a particle size of 1 microm. The model successfully depicts PM deposition patterns and their dependence on the severity of disease. The findings indicate that airway obstructions are the main cause for increased deposition in the COPD lung.     

Radionuclide imaging technologies and their use in evaluating asthma drug deposition in the lungs

Whole lung and regional lung deposition of inhaled asthma drugs in the lungs can be quantified using either two-dimensional or three-dimensional radionuclide imaging methods. The two-dimensional method of gamma scintigraphy has been the most widely used, and is currently considered the industry standard, but the three-dimensional methods (SPECT, single photon emission computed tomography; and PET, positron emission tomography) give superior regional lung deposition data and will undoubtedly be used more frequently in future. Recent developments in radionuclide imaging are described, including an improved algorithm for assessing regional lung deposition in gamma scintigraphy, and a patent-protected radiolabelling method (TechneCoat), applicable to both gamma scintigraphy and SPECT. Radionuclide imaging data on new inhaled asthma products provide a milestone assessment, and the data form a bridge between in vitro testing and a full clinical trials program, allowing the latter to be entered with increased confidence.     

Acute toxicity of intratracheal arsenic trioxide instillation in rat lungs

This study investigated the acute toxicity of different concentrations of arsenic trioxide (As₂O₃; ATO) on rat lungs. In total, 160 Wistar rats were randomly divided into the control, low‐, medium‐ and high‐dose groups, which were exposed to 0, 0.16, 1.60 and 16 μg/kg of ATO by intratracheal instillation, respectively. Samples were collected at 6, 12, 24, 48 and 72 hours after exposure and the dynamic changes indicative of acute lung toxicity were monitored. Compared with the control group, the exposure groups exhibited significant changes such as increased lung water content ratio and protein concentration in the bronchoalveolar lavage fluid, pulmonary interstitial thickening, cell membrane edema, increased inflammatory factor concentration, JNK and P38 were significantly activated, and the degree of phosphorylation was increased. Furthermore, all the changes in the exposure groups were exposure concentration‐dependent. ATO respiratory tract exposure can cause restrictive ventilatory disturbance in rats, and the degree of injury is exposure concentration‐dependent.     

Functional characterization of the muscarinic receptor in rat lungs.

The effects of various muscarinic antagonists on antigen- and acetylcholine-induced bronchoconstriction were studied. In isolated and ventilated lungs of naive rats, the pA2 values with respect to acetylcholine-induced bronchoconstriction were 9.01 (atropine), 8.39 (ipratropium bromide), 7.39 (pirenzepine), 5.94 (AF-DX 116, a M2-selective muscarinic antagonist), 6.91 (UH-AH 37, a novel muscarinic antagonist) and 9.37 (4-DAMP: 4-diphenylacetoxy-N-methylpiperidine methobromide). Except for ipratropium bromide, the slopes of the Schild plots were not significantly different from unity. None of the drugs were potent or effective in inhibiting bronchoconstriction or histamine release evoked by antigen challenge in actively sensitized rats. However, in vivo, in anesthetized spontaneously breathing rats, vagotomy and atropine (1 mg/kg) did reduce antigen-induced bronchoconstriction. It is concluded that functional muscarinic receptors in isolated rat lungs are probably of the M3 receptor subtype. With respect to antigen-induced bronchoconstriction and mediator release in a denervated model such as the isolated lung, they are of little, if any, importance. In vivo, vagotomy and atropine reduced antigen-induced bronchoconstriction, probably by blockade of a vagal reflex which is thought to play a role in antigen-evoked bronchoconstriction.     

Metabolic effects of isoflurane on rat lungs perfused in situ

1. The current experiments studied the effects of the inhalation anesthetic, isoflurane, on 5-hydroxytryptamine (5-HT) metabolism, protein synthesis, and angiotensin-converting enzyme activity in perfused rat lungs. 2. Under first order reaction conditions, isoflurane decreased the accumulation of tissue 5-hydroxyindoleacetic acid, the principle metabolite of 5-HT in a concentration-related, competitive, and reversible manner, indicating inhibition of endothelial 5-HT uptake. 3. In apparent contrast, isoflurane appeared to stimulate uptake of 5-HT by an imipramine-sensitive process, into a cell type unable to metabolize the parent amine. 4. Isoflurane increased absolute angiotensin-converting enzyme activity only at an inspired concentration of 5%. The anesthetic did not affect lung protein synthesis.     

Vinyl acetate casts of emphysematous rat lungs.

Emphysema was induced in rats by chronic exposure to a papain aerosol according to established methods. The emphysematous changes were evaluated by previously described histological and biochemical parameters and by a new method employing infiltration of the lungs with vinyl acetate to make casts of the air spaces of the pulmonary tree. The weights and gross and microscopic appearance of the casts demonstrated the emphysematous changes in lungs of the papain-treated rats. Vinyl acetate casting may offer a new and useful tool in evaluating experimental induction of emphysema in rats.     

Age-related morphometric differences in responses of rat lungs to ozone

The influence of age on morphologic changes in lungs of rats exposed to ozone was studied in female Sprague-Dawley rats, aged 60 and 444 days. Rats of both age groups were exposed continuously for 72 hr to either 0.35 or 0.80 ppm ozone, or to filtered air. Tissues were evaluated using light microscopic morphometry and scanning electron microscopy. The lungs from ozone-exposed 60-day-old rats had larger volume fractions of centriacinar lesions than lungs from exposed 444-day-old rats. Within each age group there was an observed dose response, with rats exposed to 0.80 ppm ozone having larger volume fractions of lesions than those exposed to 0.35 ppm. Only the 444-day-old rats lost body weight during the exposure period. They also had smaller fixed lung volumes than same-aged controls. All 60-day-old rats gained weight during the exposure period, although rats exposed to 0.80 ppm ozone gained less than filtered air controls. Lesions observed in both age groups of female rats were qualitatively similar to those previously described in young adult male rats. We conclude that there are age-related differences in the morphometric responses of rats to ozone exposure. Younger rats had larger proportions of centriacinar lesions and macrophages while older rats had greater body weight and lung volume changes.