Spray Administration Research Articles

Effect of Formulation- and Administration-Related Variables on Deposition Pattern of Nasal Spray Pumps Evaluated Using a Nasal Cast

To systematically evaluate the effect of formulation- and administration-related variables on nasal spray deposition using a nasal cast. Deposition pattern was assessed by uniformly coating a transparent nose model with Sar-Gel®, which changes from white to purple on contact with water. Sprays were subsequently discharged into the cast, which was then digitally photographed. Images were quantified using Adobe® Photoshop. The effects of formulation viscosity (which influences droplet size), simulated administration techniques (head orientation, spray administration angle, spray nozzle insertion depth), spray pump design and metering volume on nasal deposition pattern were investigated. There was a significant decrease in the deposition area associated with sprays of increasing viscosity. This appeared to be mediated by an increase in droplet size and a narrowing of the spray plume. Administration techniques and nasal spray pump design also had a significant effect on the deposition pattern. This simple color-based method provides quantitative estimates of the effects that different formulation and administration variables may have on the nasal deposition area, and provides a rational basis on which manufacturers of nasal sprays can base their patient instructions or post approval changes when it is impractical to optimize these using a clinical study.

Autologous Valve Replacement—CD133+ Stem Cell-Plus-Fibrin Composite-Based Sprayed Cell Seeding for Intraoperative Heart Valve Tissue Engineering

The development of biological valve prostheses with lifetime native-like performance and optimal host engraftment is an ultimate goal of heart valve tissue engineering. We describe a new concept for autologous graft coating based on a CD133(+)-stem-cells-plus-fibrin (SC+F) complex processed from bone marrow and peripheral blood of a single patient. CD133(+)-SC (1 × 10(6) cells/mL) from human bone marrow and autologous fibrin (20 mg/mL) were administered simultaneously via spray administration using the novel Vivostat Co-Delivery System. During static cultivation, SC+F performance was monitored for 20 days after delivery and compared with controls. For dynamic testing SC+F-composite was sprayed on a decellularized porcine pulmonary valve and transferred to a bioreactor under pulsatile flow conditions for 7 days. Static cultivation of SC+F-composite induced significant improvements in stem cell proliferation as compared with controls. For dynamic testing, microscopic analyses on a smooth engineered heart valve surface detected homogenous distribution of stem cells. Ultrasonic analysis revealed native-like valve performance. Applied CD133(+) stem cells differentiated into endothelial-like cells positive for CD31 and vascular endothelial growth factor receptor 2 and engrafted the valve. However, occasional delamination was observed. SC+F serves as an excellent autologous matrix for intraoperative tissue engineering of valve prostheses promising optimal in vivo integration. However, stability remains an issue.

Characterization of a novel live attenuated infectious bronchitis virus vaccine candidate derived from a Korean nephropathogenic strain

A nephropathogenic K2/01 strain of infectious bronchitis virus (IBV) was attenuated by 170 serial passages in embryonated chicken eggs for possible use as a future IBV vaccine strain. High-growth properties and narrow tissue tropisms (limited replication in respiratory tracts) were achieved by the adaptation process. Unlike the parent strain, the attenuated strain (K2p170) was safe in day-old specific-pathogen-free chicks since replication of the virus did not induce mortality and nephritis, and rarely induced histological changes in the trachea and kidney after intraocular administration. In day-old broilers, even though coarse spray administration of K2p170 induced clinical signs, ciliostasis, and histopathological lesions in the trachea and the kidney, they were all comparable to birds vaccinated with commercial H120 vaccine. Despite restriction of viral replication in the respiratory tract, K2p170 elicited the production of antiserum with a neutralization index of 4.5. K2p170 provided almost complete protection against both two distinct subgroups of Korean nephropathogenic strain (KM91-like and QX-like subgroup). Furthermore, K2p170 provided significantly greater cross-protection against two heterologous strains (Massachusetts and Korean respiratory strain) than those conferred by the commercial H120 vaccine. K2p170 also had no virulence reversion after five back passages in chickens. In conclusion, K2p170 exhibits a fine balance between attenuation and immunogenicity, possesses cross-protective efficacy, and merits further investigation as a potential live vaccine as an alternative means of protection against the recently emergent nephropathogenic IBV infection in many Eurasian countries.

Efficacy of a Spray Compound Containing a Pool of Collagen Precursor Synthetic Amino Acids (L-Proline, L-Leucine, L-Lysine and Glycine) Combined with Sodium Hyaluronate to Manage Chemo/Radiotherapy-Induced Oral Mucositis: Preliminary Data of an Open Clinical Trial

Oral mucositis (OM) is a very frequent and potentially severe complication experienced by patients receiving chemotherapy and/or radiotherapy, which often leads to significant morbidity and mortality, and decreased quality of life, and is very costly. Despite its severity and prevalence, there is no standard recognised management today. The aim of this open clinical trial is to evaluate the efficacy and compliance of a new spray compound containing sodium hyaluronate (SH) and a pool of collagen precursor amino acids (AAs) combined with sodium hyaluronate (SH) to manage radio/chemotherapy-induced OM. Twenty-seven consecutive patients with OM were treated according to the manufacturers instructions. At time T0 (baseline before intervention), we evaluated the following parameters: (i) pain score (by linear visual analogue scale; 0100) and (ii) severity of OM scored according to WHO Mucositis scale. The treatment efficacy was evaluated on i) pain score, ii) clinical resolution index (CRI) and iii) patient compliance at times T01 (after 2 hours), T1 (after 24 hours), T2 (after 72 hours), T3 (after 7 days) and T4 (after 14 days). Results showed that painful symptoms were significantly reduced after only 2 hours of spray administration compared with baseline measurements (p less than 0.0001; z=-4.541). A progressive reduction of pain through the 2 weeks was also noted (p less than 0.0001). Patient lesions treated with SH-Asbased spray also significantly improved after 72 hours of treatment (p=0.0051; z=-2.803). During the two-week observation, all patients significantly improved from the baseline (p less than 0.0001) and progressively ameliorated their ability to swallow foods and liquids. The compliance of all patients to the product was very good, and at the end of the study there were no adverse effects. The results suggest that the SHAAs-based spray accelerates lesion healing and above all helps to manage mucositis pain, especially in terms of immediate pain relief (after 2 hours from application). Although further randomized controlled studies are recommended, our findings suggest that frequent applications of this spray may offer rapid and effective pain management, aiding faster mucosal wound healing.

Citrus/Cydonia comp. for seasonal allergic rhinitis: A randomized study to compare treatment effects of the subcutaneous and the nasal routes of administration

Objectives Allergic rhinitis is a condition characterized by sneezing, watery nasal discharge, nasal obstruction and itching. It is an increasingly prevalent condition, particularly in the Western world where it affects around 20% of the adult population. Citrus/Cydonia comp. is an anthroposophic medicine, which contains extracts of lemon (Citrus lemon) and quince (Cydonia oblongata). Several observational studies have shown that Citrus/Cydonia comp. solution either as a subcutaneous injection or as a nasal spray might be an effective treatment for seasonal allergic rhinitis (SAR) patients. In addition, in vitro is has been shown that it has a selective effect on the differentiation of T-cells by producing relatively more IL-10 than IL-12. By that, it also seems to have an effect on the induction of regulatory (IL-10 producing) T-cell subsets. Therefore, it has been concluded that Citrus/Cydonia comp., at least in vitro, is capable of neutralizing (to some extent) the changes, characteristic to allergic rhinitis, with regard to the maturation, differentiation, and activity of the immune system. The aim of the present study is to compare the efficacy and safety of subcutaneous versus nasal spray administration in SAR patients by specifically investigating which route of administration is more effective in the differentiation and inductions of (regulatory) T-cells. Methods A national, randomized, comparative clinical trial with two parallel groups. A total of 28 patients with grass pollen SAR are recruited at primary care practices or through advertisements. After a 2-week run-in period without allergy medication, patients are randomized to a 6 weeks treatment period with either Citrus/Cydonia comp. 1% subcutaneous injections (2 injections per week) or Citrus/Cydonia comp. 1% nasal spray (1–2 sprays in each nostril, 4 times a day). The primary outcome variables are changes in immunological parameters such as IL-10, IL-12 and IL-5 at week 6, as studied in PBMCs isolated from the blood of the patients. Further objectives are to investigate the influence of Citrus/Cydonia comp. subcutaneous injections versus Citrus/Cydonia comp. nasal spray on the change in clinical symptoms by means of nasal and non-nasal symptom scores. The safety and tolerability of both injections and nasal spray will be investigated by determining adverse events surveillance. Results and conclusions The outcome of this randomized comparative study will be presented at the congress.

Recurrent vasovagal syncope: comparison between clomipramine and nitroglycerin as drug challenges during head-up tilt testing

To compare the responses between clomipramine, a centrally acting substance, and nitroglycerin, with mainly peripheral action, when each drug is used during tilt test for the induction of vasovagal syncope (VVS). Hundred patients with recurrent episodes of classical VVS underwent two tilt tests in a randomized sequence. One test included 20 min of tilt at 60 degrees with intravenous administration of 5 mg clomipramine (clomipramine tilt), whereas the other test included an initial 30 min period of passive 60 degrees tilt, followed by sublingual spray administration of 400 microg nitroglycerin (nitroglycerin tilt). Fifty asymptomatic subjects served as controls. Following clomipramine tilt, a positive response occurred in 73 patients (73%), a negative response in 23 (23%), and drug intolerance in 4 (4%). With nitroglycerin tilt, these percentages were 52, 48, and 0%, respectively. Significant differences were observed regarding positive responses (clomipramine vs. nitroglycerin: 73/100 vs. 52/100, P < 0.05), as well as negative responses (23/100 vs. 48/100, respectively, P < 0.05). A high concordance rate was observed in positive responses. In the evaluation of patients with recurrent classical VVS, clomipramine tilt is associated with an increased positive yield relative to nitroglycerin tilt. This suggests that central mechanisms may be more important than peripheral ones in VVS pathogenesis.

Vaccination of Chickens with Liposomal Inactivated Avian Pathogenic Escherichia coli (APEC) Vaccine by Eye Drop or Coarse Spray Administration

One hundred and twenty 30-day-old specific-pathogen-free chickens were inoculated with the liposomal inactivated avian pathogenic Escherichia coli (APEC) vaccine by eye drop or coarse spraying. All of the chickens produced anti-lipopolysaccharide antibodies of the IgG subclass in their sera as well as IgA antibodies in their oral mucus. The results demonstrated a rise in antibodies in the serum of chickens administered the APEC vaccine through nonparenteral mucosal routes. Bacterial counts in the blood decreased, and clinical signs were moderated in the vaccinated chickens after challenge with a strain of APEC. No harmful effects from the vaccination were observed. The liposomal inactivated APEC vaccine described in this paper would contribute to a practical method of control for avian colibacillosis.

Vaccination of Chickens with Lipsomal Inactivated Avian Pathogenic Escherichia coli Vaccine by Eye Drop or Coarse Spray Administration

Vaccination of Chickens with Lipsomal Inactivated Avian Pathogenic Escherichia coli Vaccine by Eye Drop or Coarse Spray Administration

Treatment of allergic rhinitis with asthma by different ways of inhaling corticosteroids

To observe the therapeutical effect on allergic rhinitis with asthma by different ways of inhaling corticosteroids. Ninety patients suffering from allergic rhinitis with asthma were classified into three groups in random and treated with budesonide (BUD) by nasal spray, inhaling and nasal spray-inhaling combined administration for 12 weeks, respectively. The concentration of serum total IgE, eosinophil cationic protein (ECP) and IL-5, pulmonary functions were examined before and after treatment. The improvement of symptom scores of rhinitis and asthma in three groups were significant (P < 0.05). The concentration of serum total IgE, ECP and IL-5 in three groups decreased greatly after treatment (P < 0.05). There were significant improvement in FEV1, FEV1/FVC and FEF25%-75% in inhaling group before and after treatment (P < 0.05): FEV1 from (2.04 +/- 0.45) L to (2.47 +/- 0.54) L, FEV1/FVC from (72.73 +/- 5.59)% to (75.42 +/- 5.94)% and FEF25%-75% from 1.69 +/- 0.52 to 2.06 +/- 0.77. There was also significant improvement in nasal spray-inhaling combined groups before and after treatment (P < 0.05): FEV1 from (2.32 +/- 0.56) L to (2.76 +/- 0.58) L, FEV1/FVC from (73.80 +/- 4.17)% to (76.04 +/- 4.49)% and FEF25%-75% from 2.09 +/- 0.45 to 2.34 +/- 0.64. The significant difference of FEV1 among the three treatment groups was observed (P = 0.041). The symptoms of rhinitis and asthma in three groups by three ways of inhaling BUD were greatly improved, no significant difference in symptom scores and serum parameters was found.

Influence of allergy in patients with nasal polyposis after endoscopic sinus surgery

Conclusion. Allergy does not modify the symptoms and steroid consumption (oral and local) of nasal polyposis (NP) patients after functional endoscopic sinus surgery (FESS). Objectives. To assess the role of allergy in the evolution after FESS of patients presenting with the diagnosis of NP. Patients and methods. This was a prospective study of 63 consecutive patients with NP (57% males, mean age 45.8 years), who were analyzed to detect whether the results of a surgical treatment of NP were influenced by the presence of positive allergic tests (Phadiatop®). Three nasal criteria were scored: nasal obstruction, posterior rhinorrhea, and the loss of smell. The frequency of asthma was evaluated. Medical treatment of NP after FESS consisted of washing of the nasal cavities, steroid spray, and oral steroid administration. The amount of consumption of steroids (prednisolone and beclomethasone) was studied. Results. Decrease of all nasal symptoms was not statistically different in the two groups of patients with and without allergy. Cumulative consumption of prednisolone and beclomethasone after surgery was similar in the two groups.

Liquid spray formulations of xibornol by using self-microemulsifying drug delivery systems

Xibornol is a lipophilic drug mainly used in Italy and Spain in spray dosage forms for the local treatment of infection and inflammation of the throat. Its poor water solubility makes difficult the development of aqueous formulations of the drug, thus giving rise to a limited number of stable and pharmaceutically accepted preparations. In fact, xibornol is actually marketed only as spray aqueous suspension. The aim of this work was to evaluate the possibility of developing a stable liquid formulation of the drug intended for oral spray administration using a self-microemulsifying drug delivery system (SMEDDS). These systems are able to adequately improve the drug solubility, allowing the introduction of relatively high concentration of drugs in the form of solution. Labrafil M1944, Labrafil M2125 and Labrafac CC were screened as oil phases, Labrasol and Labrafac PG as surfactants and Transcutol as co-surfactant. Pseudo-ternary phase diagrams were constructed, by titration with the aqueous phase of different oil phases and surfactant/co-surfactant mixtures in order to identify the self-microemulsification region and the optimal micro-emulsion composition. Then, complete pharmaceutical formulations were prepared and evaluated for stability and viscosity properties. The final selected formulations, containing Labrafil M1944, Transcutol, Labrafac PG and a hydrophilic co-solvent (propylene glycol or PEG 200) allowed complete solubilization of the required xibornol concentration (3%, w/v) and showed physical good stability up to 2 months at 25 and 4 °C, suitable viscosity and organoleptic properties.

Scopolamine Sublingual Spray: An Alternative Route of Delivery for the Treatment of Motion Sickness

ABSTRACTThe purpose of this study was to develop a sublingual drug delivery spray formulation of scopolamine hydrobromide (L-(-)-hyoscine hydrobromide) and to determine the absolute bioavailability of scopolamine hydrobromide following sublingual delivery and to investigate the effect of a bioadhesive on the pharmacokinetic parameters of this drug in a rabbit model. Rabbits received a single scopolamine free base equivalent sublingual dose of 100 μg/kg and this was compared to intravenous administration of the drug. Blood samples were collected at different time points, and plasma scopolamine concentrations were determined using a new sensitive and specific LC/MS analytical method which utilized electrospray ionization detection. The bioavailability of sublingual scopolamine was determined by comparing plasma concentrations after sublingual spray delivery with equivalent intravenous doses. Following delivery of the sublingual spray dose, the average Cmax was 1024.4 ± 177 ng/mL, and the AUC value was found to be 61067.6 ± 9605 ng.min/mL. Relative to the intravenous dose (100% bioavailability), the bioavailability was 79.8% after sublingual spray administration. The addition of 2% chitosan, a bio-adhesive material and an absorption enhancer, showed a significant improvement in scopolamine sublingual absorption (p < 0.05) was observed. Considering the limitations of delivering scopolamine orally or transdermally to patients who experience motion sickness, the sublingual route of administration using a spray delivery dosage form, is a potential alternative modality for the prevention of nausea and vomiting associated with motion sickness.

Influence of allergy on the symptoms and treatment of nasal polyposis

Conclusion. Allergy does not modify the symptoms of nasal polyposis, either initially or after a 1-year medical treatment. Objectives. To assess the role of allergy in the symptoms and treatment of patients presenting with the diagnosis of nasal polyposis. Patients and methods. Two simultaneous studies were carried out. In the first study, 180 consecutive patients with nasal polyposis (60% males, mean age = 48.4 years) were analyzed to detect whether the severity of their symptoms correlated with the presence of positive allergic tests. In the second study, 74 consecutive patients (57.5% males, mean age = 48.3 years) were analyzed to detect whether the results of a 1-year medical treatment of nasal polyposis were influenced by the presence of positive allergic tests (Phadiatop®). Five nasal criteria were scored: nasal obstruction, anterior and posterior rhinorrhea, facial pain, and the loss of sense of smell. The frequency of asthma was evaluated. Treatment of nasal polyposis consisted of washing of the nasal cavities, steroid spray, and oral steroid administration. The amount of steroid consumption (prednisolone and beclomethasone) was studied. Results. In the first study, mean scores of nasal symptoms did not differ between the two groups of patients with and without allergy. The prevalence of asthma (p = 0.03) was higher in the group with than without allergy. In the second study, decrease of all nasal symptoms was not statistically different in the two groups. Cumulative consumption of prednisolone and beclomethasone between baseline and year 1 were similar in the two groups.

True Nasopharyngeal Absorption of Zolmitriptan after Administration via Nasal Spray in Healthy Male Volunteers

The aim of this study was to quantify the fraction of zolmitriptan that is absorbed directly through the nasal mucosa after administration of zolmitriptan nasal spray. Following confirmation that activated charcoal blocks gastrointestinal absorption of zolmitriptan, healthy male volunteers were given zolmitriptan nasal spray 5mg (with and without charcoal) and zolmitriptan 5mg oral tablets (with and without charcoal) in a randomized, crossover design. Blood samples for pharmacokinetic analysis were collected up to 9 hours post-dose. Twelve subjects completed all treatments. The fraction of bioavailable dose absorbed through the nasal mucosa was calculated to be 29% (95% CI 19, 43). Intranasal absorption contributed 71% of zolmitriptan exposure during the first hour post-dose and 49% during the first 2 hours. This study confirms true nasopharyngeal absorption of zolmitriptan after nasal spray administration. This route of absorption contributed approximately 30% of total zolmitriptan exposure, with much higher proportions of exposure seen during the first 2 hours post-dose when patients would expect to benefit from treatment. These results suggest that intranasal absorption plays an important role in the early onset of efficacy of zolmitriptan nasal spray seen in clinical trials.

Pulmonary Edema following Phenylephrine Intranasal Spray Administration during the Induction of General Anesthesia in a Child

Topical phenylephrine, an agent used to facilitate nasotracheal intubation and prevent nasal mucosal bleeding, can cause severe hypertension in some patients, secondary to its stimulation of alpha-adrenergic receptors. Moreover, a high incidence of pulmonary edema is found in patients whose phenylephrine administration is followed by treatment with beta-blocking agents. We report a case of acute pulmonary edema in a pediatric patient who developed severe hypertension after the inadvertent administration of a large dose of topical nasal phenylephrine, followed by beta-adrenergic antagonists (esmolol).

Protective efficacy of a live attenuated anticoccidial vaccine administered to 1-day-old chickens

The efficacy of a live attenuated anti-coccidial vaccine, Paracox-5, administered to 1-day-old chicks was investigated by assessing protection against changes in weight gain following virulent challenge. Vaccinatedbirds were challenged independently 28 days later with each of the component species (Eimevia acervulina,Eimeria maxima, Eimeria mitis or Eimeria tenella), and protection was demonstrated against associatedreduction in weight gain and lesion formation. In addition, an improvement in bird performance, in terms of feedconversion ratio, was also observed following vaccination. Furthermore, under conditions designed to moreclosely mimic those in the field and using hatchery spray administration, protection against a mixed virulentchallenge introduced by 'seeder birds' was demonstrated evenly across a flock of broiler birds within 21 daysafter vaccination. These data demonstrate that Paracox-5 vaccine will protect broiler chickens against theadverse effects on performance induced by Eimeria spp.

Experimental eradication of pinworms (Syphacia obvelata and Aspiculuris tetraptera) from mice colonies using ivermectin.

A spray administration of ivermectin was evaluated for the treatment of pinworm infection in mice. In this study, a spray of 0.1% ivermectin injectable solution over the entire cage once a week, for three consecutive weeks (one cycle treatment), was effective in eradicating both Syphacia obvelata and Aspiculuris tetraptera from mice under experimental conditions. In addition, no acute toxicity was observed in 105 mothers or 687 neonates treated with ivermectin, indicating that ivermectin does not affect murine reproduction. Finally, we attempted to eradicate pinworms from infected mice in our institute using this method. Two cycles of treatment were administered, with a two-week pause between cycles, resulting in complete eradication for at least one year. Treating mouse colonies with spray ivermectin is inexpensive, safe, requires very little labor and is very effective at eradicating pinworms from mice.

Endothelial dysfunction associated with oxidative stress in human

The imbalance in pro- and anti-oxidant activity causes endothelial dysfunction, which precedes and exacerbates atherosclerosis. LDL-apheresis improves ischemic symptom in patients with hypercholesterolemia and regresses atherosclerosis. Whereas, patients who undergo hemodialysis experience accelerated atherosclerosis and premature death even though their cholesterol level is known to be normal. We investigated the association between oxidative modification and endothelial function during a single session of blood purification therapies, LDL-apheresis and hemodialysis. To examine the effect of a single session of LDL-apheresis on endothelial function in patients with hypercholesterolemia, we measured forearm blood flow (FBF) by strain gauge plethysmography before and after LDL-apheresis, while infusing acetylcholine (ACh) and sodium nitroprusside (SNP). The single session of LDL-apheresis reduced oxidized-LDL. Although ACh and SNP increased FBF dose-dependently before and after LDL-apheresis, ACh response was significantly augmented without changes in responses to SNP. The plasma level of oxidized LDL correlated with the degree of ACh-induced vasodilatation. To examine the effect of a single session of hemodialysis on endothelial function in patients with end stage renal disease, we estimated flow-mediated vasodilation (FMD) during reactive hyperemia using a high-resolution ultrasonography, before and after a single session of hemodialysis. We also investigated the effect of anti-oxidative modification during hemodialysis using an a-tocopherol acetate-coated or a non-coated cellulose dialyzer. Non-specific endothelium independent vasodilation was measured after sublingual glyceryltrinitrate spray (GTN; 0.3 mg) administration in each session. A single session of hemodialysis increased plasma level of oxidized LDL. Although hemodialysis treatment did not affect GTN-induced vasodilation, a single session of hemodialysis impaired the endothelium-dependent vasodilation. Treatment with vitamin E-coated-dialyzer restored endothelium-dependent vasodilation, which was accompanied by decrease in oxidized LDL. Thus, endothelial function was impaired by acute increase in oxidative stress and was restored by anti-oxidant therapy. Our results suggest the importance of controlling oxidative stress in addition to classical risk factors to preserve human endothelial function.

Évaluation in vitro et in vivo d’une chambre d’inhalation fabriquée en Tunisie

Aerosol delivered through metered-dose inhalers offers an efficacious way to deliver aerosols to asthmatics. Unfortunately, children and aged subjects are unable to coordinate satisfactory their use with inhalation, a problem overcome by using spacer devices. Purpose. – To compare, in vitro and in vivo, a new Tunisian spacer device: Airbox® (Omezine Laboratory) with Volumatic® (Glaxo Wellcome). Methods. – The experimental model includes an artificial ventilator, which simulates a seven-year-old child’s respiration, connected to spacer devices. A millipore filter was placed in serie at the exit of the spacer. The filter was weighed blind by a high precision balance (± 0.01 mg) before and after the spray administration. Thirty-six children aged from 6 to 14 years were enrolled in a randomised, double-blind, crossover, in vivo study. All children have severe persistent asthma with forced expiratory volume in a one-second (FEV 1) baseline between 50 and 60% of reference value. The reversibility of airway obstruction was assessed by the FEV 1 and the change after inhalation of 200 μg of salbutamol delivered via each spacer devices. A serie of 30 experiments was made with different doses of salbutamol. Results. – The amount of aerosol retained by the filter was statically similar for both spacer devices. Reversibility percentages measured by the two spacer devices was in the same confidence interval. Conclusions. – The salbutamol-Airbox® association was as efficient as the salbutamol-Volumatic® association in vitro and in vivo.

Host responses and persistence of vector genome following intrabronchial administration of an E1(-)E3(-) adenovirus gene transfer vector to normal individuals.

Adenovirus (Ad)-mediated gene transfer to the respiratory epithelium of experimental animals and to nasal and airway epithelium of individuals with cystic fibrosis is followed by transient gene expression. Extensive studies in experimental animals are consistent with the concept that local cellular host anti-vector immune responses account for this short-term expression, and systemic and local [lung epithelial lining fluid (ELF)] anti-Ad neutralizing antibodies are generated following Ad vector administration to the respiratory epithelial surface. To determine if this paradigm holds in normal humans, a first-generation Ad vector (Ad(GV)CD.10, an E1(-)E3(-) Ad serotype 5-based vector coding for the Escherichia coli cytosine deaminase gene) was sprayed locally in escalating doses (8 x 10(8)-8 x 10(10) particle units (pu), n = 2/group) into the lung airway epithelium of six normal individuals. Serum, ELF, and endobronchial biopsies were obtained at baseline and at various time points following vector administration. In contrast to the observations in experimental animals in which lung administration of first-generation Ad vectors is followed by strong systemic and local host response, bronchial spray administration of the Ad vector to normal humans showed: (1) minimal inflammation in bronchial biopsies, bronchial brushing, and bronchoalveolar lavage fluid; (2) no blood lymphocyte proliferation in five of six individuals in response to in vitro stimulation with Ad antigens; and (3) no significant increase from baseline in blood or lung ELF anti-Ad neutralizing antibodies. Despite this minimal normal human anti-Ad host response, dose-dependent levels of vector DNA in the airway epithelium were transient. Vector DNA in the targeted airway epithelial cells peaked in a dose-dependent fashion at 0.007 to 1.1 copies/cell at day 7 and declined thereafter, reducing to <10% of peak levels by 2 weeks. These observations demonstrate both the strengths and the limits of using experimental animals to predict human responses to gene transfer vectors. While the transient nature of Ad vector persistence in the airway epithelium is predicted by most experimental animal studies, respiratory epithelial administration of first-generation Ad vectors at doses up to 10(10) pu to airway epithelium of healthy individuals elicits minimal to no detectable systemic and mucosal humoral and cellular immune responses, an observation diametrically opposed to the host responses measured in experimental animals. These findings suggest that, while adaptive anti-Ad immune responses likely play some role in the disappearance of the vector DNA following vector administration to the human lung, other mechanisms may also be involved in the response of humans to Ad gene transfer vectors.