Changes In Barrier Properties Research Articles

Determination of Endothelial Barrier Resistance by Electric Cell-Substrate Impedance Sensing (ECIS) System.

Endothelial cells lining the inner surface of blood vessels and lymphatic vessels play an indispensable role in vascular homeostasis. Apart from regulating vessel tone and forming an anti-thrombotic and anti-atherosclerotic surface, the dynamic endothelial barrier controls transport of solutes and fluid in and out of tissues at the capillary bed. Transit of circulating leukocytes into and out of circulation during inflammation and tissue repair is also regulated by the endothelium. Dysregulation of this barrier function of endothelial cells is a hallmark feature of multiple diseases and conditions such as sepsis, cancer metastasis, and edema. In this chapter we describe a detailed methodology to perform an in vitro experiment to monitor changes in barrier properties of human umbilical vein endothelial cells (HUVECs) in real time, in response to thrombin with electrical cell-substrate impedance sensing (ECIS) biosensor system.

Characterization of Biodegradable Food Contact Materials under Gamma-Radiation Treatment.

Radiation is an example of one of the techniques used for pasteurization and sterilization in various packaging systems. There is a high demand for the evaluation of the possible degradation of new composites, especially based on natural raw materials. The results of experimental research that evaluated the impact of radiation technology on biodegradable and compostable packaging materials up to 40 kGy have been presented. Two commercially available flexible composite films based on aliphatic-aromatic copolyesters (AA) were selected for the study, including one film with chitosan and starch (AA-CH-S) and the other with thermoplastic starch (AA-S). The materials were subjected to the influence of ionizing radiation from 10 to 40 kGy and then tests were carried out to check their usability as packaging material for the food industry. The results showed that the mechanical properties of AA-S films improved due to the radiation-induced cross-linking processes, while in the case of AA-CH-S films, a considerable decrease in the elongation at break was observed. The results also showed a decrease in the WVTR in the case of AA-S and no changes in barrier properties in the case of AA-CH-S. Both materials revealed no changes in the odor analyzed by sensory analysis. In the case of the AA-S films, the higher the radiation dose, the faster the biodegradation rate. In the case of the AA-CH-S film, the radiation did not affect biodegradation. The performed research enables the evaluation of the materials intended for direct contact with food. AA-CH-S was associated with unsatisfactory parameters (exceeding the overall migration limit and revealing color change during storage) while AA-S showed compliance at the level of tests carried out. The study showed that the AA-CH-S composite did not show a synergistic effect due to the presence of chitosan.

Induction of permeability and apoptosis in colon cancer cell line with chitosan

The effect of chitosan on tight junctions (TJs) permeability on Caco-2 cell monolayer intestinal model was investigated. We have also investigated the effects of low molecular weight water-soluble chitosan (LMWC) on apoptosis in HT-29 cell. The changes in barrier properties of Caco-2 cell monolayers, including transepithelial electrical resistance (TEER) and permeability to Lucifer yellow, were assessed in response to chitosan treatment. High molecular weight chitosan (HMWC) was found to cause up to 50% dose-dependent reduction in TEER of Caco-2 cell monolayer without damage to the cell membrane under lower concentration. The effect of HMWC on TJs was confirmed by increased permeability of lucifer yellow when cells were treated with 0.001-0.0001% HMWC for 120 min compared to control cells. Results showed that HMWC did not affect the F-actin of cytoskeleton. LMWC was proven to be an antitumor compound as shown by inducing apoptosis as a function of DNA fragmentation. These results suggest that HMWC is an useful drug delivery agent in paracellular pathway and LMWC has potential value in colon cancer therapy.

Development of an Oxygen Sensitive Model Gel System to Detect Defects in Metal Oxide Coated Multilayer Polymeric Films

Metal oxide coated multilayered polymeric pouches provide a suitable alternative to foil-based packaging for shelf-stable products with extended shelf-life. The barrier performance of these films depends upon the integrity of the metal oxide coating which can develop defects as a result of thermal processing and improper handling. In this work, we developed a methodology to visually identify these defects using an oxygen-sensitive model gel system. Four pouches with different metal oxide coatings: MOA (Coated PET), MOB (SiOx -coated PET), MOC (Overlayer-AlOx -Organic-coated PET), MOD (Overlayer-SiOx -coated PET) were filled with water and retort-processed for 30 and 40 min at 121 °C. After processing, the pouches were cut open, dried and subsequently filled with a gel containing methylene blue that changes color in the presence of oxygen. The pouches were then stored at 23 and 40 °C for 180 and 90 days, respectively. Defects were identified by observing the localized color change from yellow to blue in the packaged gel. These observations were confirmed through measurement of oxygen and water vapor transmission rates, as well as SEM and CLSM analyses. The MOC pouches showed the least change in barrier properties after thermal processing. This was due to crosslinking in the organic coating and protection provided by the overlayer. The melting enthalpy of all films increased significantly (P < 0.05) after sterilization. This may increase the brittleness of the substrates after processing. Findings may be used to improve the barrier performance of metal oxide coated polymeric films intended for food packaging applications. PRACTICAL APPLICATION: In this study, we developed a methylene blue-based, oxygen-sensitive model gel system to identify defects in metal oxide coated polymeric structures induced by thermal processing and mechanical stresses. We also performed a comprehensive analysis of these defects through CLSM and SEM. The gel system and methodology developed may be useful in the design and development of high barrier metal oxide coated films.

Application of Nanofibrillated Cellulose on BOPP/LDPE Film as Oxygen Barrier and Antimicrobial Coating Based on Cold Plasma Treatment

The application of nanofibrillated cellulose (NC) films in packaging industry has been hindered by its lack of heat-sealing ability. Incorporation of NC films with the biaxially oriented polypropylene/low density polyethylene (BOPP/LDPE) laminates can take advantage of each material and endow the films with novel functions for food packaging applications. In this study, a coating that consists of NC and nisin was applied onto a cold plasma treated BOPP/LDPE film to fabricate a novel active packaging with an improved oxygen barrier performance and an added antimicrobial effect. The results showed that cold plasma treatment improved the surface hydrophilicity of BOPP/LDPE films for better attachment of the coatings. NC coatings significantly enhanced oxygen barrier property of the BOPP/LDPE film, with an oxygen transmission rate as low as 24.02 cc/m2·day as compared to that of the non-coated one (67.03 cc/m2·day). The addition of nisin in the coating at a concentration of 5 mg/g caused no significant change in barrier properties but imparted the film excellent antimicrobial properties, with a growth inhibition of L. monocytogenes by 94%. All films exhibit satisfying mechanical properties and transparency, and this new film has the potential to be used as antimicrobial and oxygen barrier packaging.

Linking morphology changes to barrier properties of polymeric packaging for microwave‐assisted thermal sterilized food

ABSTRACTIt is a priority to develop polymeric packaging that can withstand microwave‐assisted thermal sterilization (MATS) and maintain the quality of low‐acid foods during long‐term storage. In this study, we explored changes in the morphology of pouch films with two multi‐layer structures. The films are based on barrier layers of metal oxide‐coated poly(ethylene terephthalate) (PET) (film A) and ethylene vinyl alcohol (EVOH) (film B). A 8‐oz model food in pouches was processed with MATS (F0 = 9.0 min) and stored at 23, 35 and 45 °C for up to 12 months. Findings reveal that the oxygen barrier of film A was influenced by the coating and crystallinity of PET. The oxygen barrier of film B was primarily affected by the moisture content of the EVOH polymer. Results also show that changes in barrier properties depended on storage temperature. Recrystallization in polymer might be an important morphological change that occurs during storages. © 2017 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2017, 134, 45481.

Detection of charge density wave phase transitions at 1T-TaS2/GaAs interfaces

The transition metal dichalcogenide 1T-TaS2 is well known to harbor a rich variety of charge density wave (CDW) distortions which are correlated with underlying lattice atom modulations. The long range CDW phases extend throughout the whole crystal and terminate with charge displacements at the crystal boundaries. Here, we report on the transport properties and capacitance characteristics of the interface between freshly exfoliated flakes of 1T-TaS2 in intimate van der Waals contact with n-type GaAs substrates. The extracted barrier parameters (ideality, barrier height, and built-in potential) experience pronounced changes across the Mott-CDW transition in the 1T-TaS2. The CDW-induced changes in barrier properties are well described by a bond polarization model which upon decreasing temperature gives rise to an increased potential drop across the interfacial region due to the localization of carriers and a decreased dielectric constant.

Minimizing Corrosion of Outdoor Metalworks Using Dispersed Chemically Stabilized Nanoclays in Polyvinylidene Fluoride Latex Coatings.

Nanoclays are small enough to appear optically transparent, yet they have large surface-to-volume and high aspect ratios that can significantly inhibit water diffusion when incorporated into protective coatings. Clear coatings, which minimally affect the aesthetics of metalworks, are commonly applied to outdoor metalworks, such as sculptures, to prevent and slow corrosion. In recent years, waterborne clear coatings, rather than solvent-based clear coatings, are increasingly used in many applications to reduce the quantity of volatile organic components in the formulation, yet the performance of dry films produced from waterborne colloidal suspensions is generally poorer. In this work, we aim to improve the barrier properties of a highly weatherable waterborne acrylic/polyvinylidene fluoride emulsion by adding a synthetic nanoclay, Laponite, into the formulation. To improve clay–polymer compatibility, the clay was covalently modified using an acetoxy or perfluoroalkyl silane monomer that is reactive with the hydroxyl groups at the edges of the Laponite platelets. Cation exchange on the clay faces using phosphorylcholine was conducted to increase the stability in water and characterized by zeta potential. Resulting changes in barrier properties of the polymer nanocomposite films were characterized by gravimetry, colorimetry, and electrochemical impedance spectroscopy. Surface ablation after accelerated artificial weathering was monitored by attenuated total internal reflectance Fourier transform infrared microspectroscopy and Raman microspectroscopy, thin film X-ray diffraction (TF-XRD) and gloss and thickness measurements. The composite films showed many improved properties: reduced water sensitivity and ultraviolet-induced polymer degradation, which increased the barrier properties and reduced the diffusion constants over both short- and long-term weathering studies compared with films without nanoclays. The diffusion constant measured for the highest performing composite film showed that the performance gap between relevant water- and solvent-borne coatings used to protect outdoor metals was narrowed by half.

The study of air-plasma treatment on corn starch/poly(ε-caprolactone) films

In spite of usefulness of synthetic polymers in every aspects of life, the environmental hazards limit their use. Starch based biodegradable polymers is one of the solutions to it. Packaging has a major share in use of synthetic polymers. For packaging application, it is necessary to have good surface and barrier properties of the material. Plasma surface modification of materials is a promising solution to enhance surface properties. In the present paper, cornstarch/poly(ε-caprolactone) (CSPCL) films were treated in air-plasma for different durations of time. The effect of air-plasma treatment on surface properties and biodegradation was studied. The extent of etching was evaluated from weight change (%) study. Changes in surface chemical composition were analyzed using ATR-FTIR and XPS. The contact angle and surface free energy (SFE) study indicate that air-plasma treatment leads to hydrophilization of CSPCL films. The changes in surface topography of plasma processed films were analyzed using AFM and SEM. The roughness caused by etching and increase in surface free energy facilitates the improvement in adhesive properties like printability and peel strength. Changes in barrier properties were studied using water vapor and oxygen transmission rate. Effect of air-plasma treatment on biodegradation of treated and untreated samples was studied by simulating natural biodegradation conditions in a controlled environment using indoor soil burial method and with a single bacterial system comprising of a commonly occurring soil bacterium, Bacillus subtilis MTCC 121. While the soil system is indicative of biodegradation due to macro as well as micro elements, a single microbial system will identify the interaction between the microorganisms and modified surface thus showing the effect of air-plasma treatment on the degradation process. Biodegradation by indoor soil burial method was assessed by measuring loss in tensile properties and growth of soil micro flora on surface by optical light microscopy (OLM). Biodegradation by B. subtilis was assessed by measuring increase in its number along with the changes it brought about in the sample surface by optical light microscopy and SEM. It was observed that such surface modifications enhanced the biodegradation rate along with finding application in packaging field, thus providing a green solution for the increasing packaging utilization and addressing environmental concerns.

Impedance analysis of GPCR-mediated changes in endothelial barrier function: overview and fundamental considerations for stable and reproducible measurements.

The past 20 years has seen significant growth in using impedance-based assays to understand the molecular underpinning of endothelial and epithelial barrier function in response to physiological agonists and pharmacological and toxicological compounds. Most studies on barrier function use G protein-coupled receptor (GPCR) agonists which couple to fast and transient changes in barrier properties. The power of impedance-based techniques such as electric cell-substrate impedance sensing (ECIS) resides in its ability to detect minute changes in cell layer integrity label-free and in real-time ranging from seconds to days. We provide a comprehensive overview of the biophysical principles, applications, and recent developments in impedance-based methodologies. Despite extensive application of impedance analysis in endothelial barrier research, little attention has been paid to data analysis and critical experimental variables, which are both essential for signal stability and reproducibility. We describe the rationale behind common ECIS data presentation and interpretation and illustrate practical guidelines to improve signal intensity by adapting technical parameters such as electrode layout, monitoring frequency, or parameter (resistance versus impedance magnitude). Moreover, we discuss the impact of experimental parameters, including cell source, liquid handling, and agonist preparation on signal intensity and kinetics. Our discussions are supported by experimental data obtained from human microvascular endothelial cells challenged with three GPCR agonists, thrombin, histamine, and sphingosine-1-phosphate.

Ultrastructural changes in the equine colonic mucosa after ischaemia and reperfusion

Ultrastructural changes in the epithelium can provide information on early changes in barrier properties, repair and inflammation in equine colon after ischaemia and reperfusion (I/R). To describe the morphology and ultrastructure of the epithelium in equine large colonic mucosa after I/R, and the response of inflammatory cells to injury. Ischaemia was induced for 1 h followed by 4 h of reperfusion in a 40 cm segment of the pelvic flexure in 6 horses. Mucosal biopsies before and after ischaemia, and after 1, 2 and 4 h of reperfusion were fixed in glutaraldehyde/paraformaldehyde and osmium tetroxide, and embedded in epon. Morphological and ultrastructural changes were evaluated in toluidine blue-stained semithin sections by light microscopy and in thin sections stained with uranyl acetate/lead citrate by transmission electron microscopy. Ischaemia caused swelling of epithelial cells and their organelles, opening of tight junctions, detachment from the basement membrane, early apoptosis and single cell necrosis. Autophagy was a prominent feature in epithelial cells after ischaemia. Reperfusion was characterised by apoptosis, epithelial regeneration and restoration of apical cell junctions. Phagocytic-like vacuoles containing cellular debris and bacteria were evident in epithelial cells after reperfusion. Paracellular and subepithelial clefts formed, accompanied by infiltration of neutrophils, lymphocytes and eosinophils into the epithelium. Subepithelial macrophages and luminal neutrophils had increased phagocytic activity. Ischaemia caused ultrastructural damage to the colonic epithelium, but epithelial cells recovered during reperfusion. Transmission electron microscopy can demonstrate subtle ultrastructural damage to epithelial cells and evidence of recovery after I/R in equine colon.

Occludin is required for cytokine-induced regulation of tight junction barriers

The function of occludin remains elusive. Proposed roles include maintenance of tight junction barriers, signaling and junction remodeling. To investigate a potential role in mediating cytokine-induced changes in barrier properties, we measured barrier responses to interferon-gamma plus TNFalpha in control, occludin-overexpressing and occludin knockdown MDCK II monolayers. MDCK cells show a complex response to cytokines characterized by a simultaneous increase in the transepithelial electrical resistance and a decrease in the barrier for large solutes. We observed that overexpression of occludin increased and occludin knockdown decreased sensitivity to cytokines as assessed by both these parameters. It is known that caveolin-1 interacts with occludin and is implicated in several models of cytokine-dependent barrier disruption; we found that occludin knockdown altered the subcellular distribution of caveolin-1 and that partitioning of caveolin into detergent-insoluble lipid rafts was influenced by changing occludin levels. Knockdown of caveolin decreased the cytokine-induced flux increase, whereas the increase in the electrical barrier was unaltered; the effect of double knockdown of occludin and caveolin was similar to that of occludin single knockdown, consistent with the possibility that they function in the same pathway. These results demonstrate that occludin is required for cells to transduce cytokine-mediated signals that either increase the electrical barrier or decrease the large solute barrier, possibly by coordinating the functions of caveolin-1.

Development of an experimental technique to assess the permeability of metal coated polymer films

The experimental time required to measure water vapour permeability of barrier film using the cup method is extremely long. In this study, a new technique is proposed, based on light transmission. This fast and accurate method was first validated using model films constituted of aluminized PET (polyethylene terephthalate) with aluminium layers of different thicknesses (<100 nm). It was possible to show that the ‘illuminating’ method could advantageously be employed to control the film’s barrier properties during their fabrication. In a second step, the method was tentatively used to measure the changes in barrier properties over time, when the samples are submitted to severe hydrothermal ageing (70 °C, 90% relative humidity [RH]). It was concluded that the complex degradation mechanism prevents accurate measurement while ageing.

Review: Role of developmental inflammation and blood–brain barrier dysfunction in neurodevelopmental and neurodegenerative diseases

The causes of most neurological disorders are not fully understood. Inflammation and blood-brain barrier dysfunction appear to play major roles in the pathology of these diseases. Inflammatory insults that occur during brain development may have widespread effects later in life for a spectrum of neurological disorders. In this review, a new hypothesis suggesting a mechanistic link between inflammation and blood-brain barrier function (integrity), which is universally important in both neurodevelopmental and neurodegenerative diseases, is proposed. The role of inflammation and the blood-brain barrier will be discussed in cerebral palsy, schizophrenia, Parkinson's disease, Alzheimer's disease and multiple sclerosis, conditions where both inflammation and blood-brain barrier dysfunction occur either during initiation and/or progression of the disease. We suggest that breakdown of normal blood-brain barrier function resulting in a short-lasting influx of blood-born molecules, in particular plasma proteins, may cause local damage, such as reduction of brain white matter observed in some newborn babies, but may also be the mechanism behind some neurodegenerative diseases related to underlying brain damage and long-term changes in barrier properties.

Dynamic Changes in the Cervical Epithelial Tight Junction Complex and Differentiation Occur during Cervical Ripening and Parturition

Cervical epithelia have numerous functions that include proliferation, differentiation, maintenance of fluid balance, protection from environmental hazards, and paracellular transport of solutes via tight junctions (TJs). Epithelial functions must be tightly regulated during pregnancy and parturition as the cervix undergoes extensive growth and remodeling. This study evaluated TJ proteins, as well as markers of epithelial cell differentiation in normal and cervical ripening defective mice to gain insights into how the permeability barrier is regulated during pregnancy and parturition. Although numerous TJ proteins are expressed in the nonpregnant cervix, claudins 1 and 2 are temporally regulated in pregnancy. Claudin 1 mRNA expression is increased, whereas claudin 2 expression declines. The cellular localization of claudin 1 shifts at the end of pregnancy (gestation d 18.75) to the plasma membrane in a lattice pattern, consistent with TJs in the apical cells. The timing of claudin 1-enriched TJs coincides with initiation of terminal differentiation of cervical squamous epithelia as evidenced by the increased expression of genes by differentiated epithelia late on gestation d 18. The cervical ripening defective steroid 5alpha-reductase type 1 deficient mouse, which has an elevated local progesterone concentration, also has aberrant claudin 1 and 2 expressions, fails to form claudin 1-enriched TJs, and lacks normal expression of genes involved in epithelial terminal differentiation. These data suggest that changes in permeability barrier properties during cervical ripening are, in part, negatively regulated by progesterone, and that dynamic changes in barrier properties of the cervix occur during pregnancy and parturition.

In vitro and in vivo evaluation of a novel oral insulin formulation

To develop a stable self-emulsifying formulation for oral delivery of insulin. Caco-2 cell line and diabetic beagles were used as in vitro and in vivo models to study the absorption mechanism and the hypoglycemic efficacy of the formulation. In addition, various physicochemical parameters of the formulation such as droplet size, insulin encapsulation efficiency and stability were evaluated. This formulation enabled changes in barrier properties of Caco-2 monolayers, as referred by transepithelial electrical resistance (TEER) and apparent permeability coefficients (P(app)) of the paracellular marker ranitidine (20-fold greater than control) but not transcellular marker propranolol, suggesting that the opening of tight junctions was involved. In diabetic beagle dogs, the bioavailability of this formulation was up to 15.2% at a dose of 2.5 IU/kg in comparison with the hypoglycemic effect of native insulin (0.5 IU/kg) delivered by subcutaneous injection. This formulation, recently approved by the China State Food and Drug Administration to enter clinical trials, was stable, degradation-protected and absorption-enhanced, and provided a promising formulation for oral insulin delivery.

Corrosion-induced copper runoff from naturally and pre-patinated copper in a marine environment

Corrosion-induced copper runoff has been monitored for copper sheet, naturally patinated copper and pre-patinated copper, with and without surface treatments, in a marine environment during one year. The study comprises solution measurements on total copper runoff rates, sulphates and chlorides released from the patina, and parallel surface analytical studies on patina formation, combined with electrochemical impedance measurements on changes in barrier properties during exposure. Bioassay tests and model predictions were applied to elucidate copper bioavailability at the immediate release situation. The runoff rate of copper was significantly lower compared to the corrosion rate throughout the exposure period. At comparable rain quantities, copper runoff rates were significantly lower at the marine site compared to similar data obtained in an urban environment. The bioavailable concentration of released copper was significantly lower compared to the total copper concentration.

Effect of self-microemulsifying drug delivery systems containing Labrasol on tight junctions in Caco-2 cells

The aim of this study was to investigate the effect of two novel self-microemulsifying drug delivery systems (SMEDDS) containing Labrasol with different dilutions on tight junctions. Changes in barrier properties of Caco-2 cell monolayers, including transepithelial electrical resistance (TEER) and permeability to the paracellular marker, i.e., mannitol, were assessed in response to dilutions and surfactants contents within formulations. The cytotoxicity of SMEDDS and the effect of surfactants on Caco-2 cells were evaluated by the MTT. Changes in subcellular localization of the tight junction proteins, ZO-1 and F-actin, were examined by confocal laser scanning microscopy. Results demonstrated that negatively charged SMEDDS with different dilutions had no effect on the TEER, but significantly increased the permeability of mannitol. In contrast, the positively charged formulation showed a dilution-dependent reduction in TEER. A corresponding increase in mannitol permeability of up to 29.4-fold to 64.7-fold greater than the control was also observed across the monolayer. Labrasol with the concentration of 0.1 and 1% was shown to increase the permeability of mannitol by 4.6-fold and 33.8-fold, respectively. The mechanism of opening of tight junctions was found to involve F-actin-related changes and redistribution of ZO-1.

Effect of Chitosan on Epithelial Cell Tight Junctions

Chitosan has been proposed as a novel excipient for transepithelial drug-delivery systems. Chitosan is thought to disrupt intercellular tight junctions, thus increasing the permeability of an epithelium. The effect of chitosan on tight junction complex was investigated at the molecular level. Changes in barrier properties of Caco-2 cell monolayers, including transepithelial electrical resistance and permeability to horseradish peroxidase (HRP), were assessed in response to chitosan treatment. Changes in subcellular localization of the tight junction proteins zona occludens 1 (ZO-1) and occludin by immunofluorescence and Western blotting of cellular fractions were also assessed. Chitosan was found to cause a dose-dependent reduction in transepithelial electrical resistance of Caco-2 monolayers of up to 83%. A corresponding increase in horseradish peroxidase permeability of up to 18 times greater than the control was also observed across the monolayer. Immunofluorescent localization of ZO-1 revealed loss of membrane-associated ZO-1 from discrete areas. Analysis of cellular fractions revealed a dose-dependent loss of ZO-1 and occludin from the cytosolic and membrane fractions into the cytoskeletal fraction. These changes did not occur because of chitosan-mediated ATP depletion. Chitosan-mediated tight junction disruption is caused by a translocation of tight junction proteins from the membrane to the cytoskeleton.

Large porous particle impingement on lung epithelial cell monolayers--toward improved particle characterization in the lung.

The ability to optimize new formulations for pulmonary delivery has been limited by inadequate in vitro models used to mimic conditions particles encounter in the lungs. The aim is to develop a physiologically-relevant model of the pulmonary epithelial barrier that would allow for quantitative characterization of therapeutic aerosols in vitro. Calu-3 human bronchial epithelial cells were cultured on permeable filter inserts under air-interfaced culture (AIC) and liquid-covered culture (LCC) conditions. Calu-3 cells grown under both conditions formed tight monolayers and appeared physiologically similar by SEM and immunocytochemical staining against cell junctional proteins and prosurfactant protein-C. Aerosolized large porous particles (LPP) deposited homogeneously and reproducibly on the cell surface and caused no apparent damage to cell monolayers by SEM and light microscopy. However, monolayers initially grown under LCC conditions showed a significant decrease in barrier properties within the first 90 min after impingement with microparticles, as determined by transepithelial electrical resistance (TEER) measurements and fluorescein-sodium transport. Conversely, AIC grown monolayers showed no significant change in barrier properties within the first 90 min following particle application. A dense mucus coating was found on AIC grown Calu-3 monolayers, but not on LCC grown monolayers, which may protect the cell surface during particle impinging. This in vitro model, based on AIC grown Calu-3 cells, should allow a more relevant and quantitative characterization of therapeutic aerosol particles intended for delivery to the tracheobronchial region of the lung or to the nasal passages. Such characterization is likely to be particularly important with therapeutic aerosol particles designed to provide sustained drug release in the lung.