Changes In Barrier Function Research Articles

Disruption of barrier function in dermatophytosis and pityriasis versicolor

Dermatophytes have the ability to form molecular attachments to keratin and use it as a source of nutrients, colonizing keratinized tissues, including the stratum corneum of the skin. Malassezia species also affect the stratum corneum of the skin. Therefore, dermatophytosis and pityriasis versicolor of the skin are thought to be important factors of profound changes in skin barrier structure and function. We aimed to describe the changes in transepidermal water loss (TEWL), stratum corneum hydration, and skin pH in the lesions of the dermatophytosis and pityriasis versicolor. Thirty-six patients with dermatophytosis (14 with tinea cruris, 13 with tinea corporis and nine with tinea pedis or tinea manus) and 11 patients with pityriasis versicolor were included in this study. TEWL, stratum corneum conductance and skin pH were determined by biophysical methods to examine whether our patients exhibited changes in barrier function. Dermatophytosis and pityriasis versicolor except tinea pedis and tinea manus showed highly significant increase in TEWL compared with adjacent infection-free skin. Hydration was significantly reduced in lesional skin compared with adjacent infection-free skin. From this study, infections with dermatophytes and Malassezia species on the body can alter biophysical properties of the skin, especially the function of stratum corneum as a barrier to water loss. On the contrary, infections with dermatophytes on the palms and soles little affect the barrier function of the skin.

Efficacy of Combined Peroxisome Proliferator-Activated Receptor-α Ligand and Glucocorticoid Therapy in a Murine Model of Atopic Dermatitis

Although topical glucocorticoids (GCs) display potent anti-inflammatory activity in inflamed skin, they also can exert numerous harmful effects on epidermal structure and function. In contrast, topical applications of ligands of peroxisome proliferator-activated receptor-α (PPARα) not only reduce inflammation, and also improve cutaneous barrier homeostasis. Therefore, we examined whether sequential topical GCs followed by topical Wy14643 (a ligand of PPARα) might be more effective than either alone for atopic dermatitis (AD) in a hapten (oxazolone)-induced, murine model with multiple features of AD (Ox-AD). Despite expected anti-inflammatory benefits, topical GC alone induced: i) epidermal thinning; ii) reduced expression of involucrin, loricrin and filaggrin; and iii) allowed outside-to-inside penetration of an epicutaneous tracer. While Wy14643 alone yielded significant therapeutic benefits in mice with mild or moderate Ox-AD, it was less effective in severe Ox-AD. Yet, topical applications of Wy14643 after GC was not only significantly effective comparable to GC alone, but it also prevented GC-induced structural and functional abnormalities in permeability barrier homeostasis. Moreover, rebound flares were largely absent after sequential treatment with GC and Wy14643. Together, these results show that GC and PPARα ligand therapy together is not only effective but also prevents development of GC-induced side effects, including rebound flares, in murine AD.

Impedance-Based Surveillance of Transient Permeability Changes in Coronary Endothelial Monolayers after Exposure to Ionizing Radiation

The relative radiation sensitivities of the various compartments of the heart are poorly characterized. Cardiac fibrosis is a common side effect of radiotherapy, suggesting that endothelial barrier function is an important factor in radiation-induced pathology. We employed Electric Cell Substrate Impedance Sensing (ECIS) to assess cytoskeletal rearrangement, permeability changes and endothelial barrier function changes in response to radiation in studies of human coronary arterial endothelial cells (HCAECs). A 5-Gy dose of γ radiation resulted in a significant sixfold transient decrease in transmonolayer resistance 3 h postirradiation (P = 0.001). This decrease in resistance coincided with changes in fluorescent tracer flux (P = 0.05) and display of an actin bundling phenotype. After irradiation, decreases in wound healing (P = 0.03) and micromotion within the monolayer (P = 0.02) were also observed. Time-lapse video studies confirmed that the monolayer is dynamic and showed that cells are extruded from the monolayer at a higher frequency after irradiation. These findings suggest that perturbed endothelial barrier function in the heart can occur at lower doses of γ radiation than previously reported.

Clinical application of the ocular fluorophotometer

The purpose of this article is to discuss the literature on the clinical role of the ocular fluorophotometer in different ocular conditions. The ocular fluorophotometer measures the fluorescent compounds present in the cornea, lens, vitreous body and retinal pigment epithelium. These compounds can be present naturally, known as autofluorescent, or fluorescent after application of fluorescein either topically or systemically. The fluorophotometer measures the changes in barrier function in both corneal epithelium and endothelium. It is a quantitative method of detecting a breakdown in the blood–ocular barriers. It helps to measure the inward and outward movements of fluorescein across the blood–retinal barrier as well as blood–aqueous barrier permeability. The measurement of ocular barrier permeability by the fluorophotometer helps to improve our understanding of ocular conditions. It is also useful to assess the effect specific therapeutic approaches in some diseases and clinical research.

Septin-2 Mediates Airway Epithelial Barrier Function in Physiologic and Pathologic Conditions

Epithelial cells have the ability to regulate paracellular permeability dynamically in response to extracellular stimuli. With every respiratory effort, airway epithelial cells are exposed to both physiologic as well as pathologic stimuli, and regulation of the epithelial barrier in response to these stimuli is crucial to respiratory function. We report that increased membrane septin-2 localization mediates decreases in paracellular permeability by altering cortical actin arrangement in human airway epithelial cells. This phenomenon occurs in response to both physiologic levels of shear stress and a pathologic stimulus, particular matter exposure. The resulting changes in barrier function in response to septin-2 redistribution have a significant impact on the ability of the apical ligand, epidermal growth factor, to interact with its receptor, epidermal growth factor receptor, which is segregated to the basolateral side in airway epithelial cells. This suggests that the dynamic regulation of the epithelial barrier function is essential in regulating signaling responses to extracellular stimuli. These findings indicate that septin-2 plays a fundamental role in regulating barrier function by altering cortical actin expression.

Extracorporeal Membrane Oxygenation Causes Loss of Intestinal Epithelial Barrier in the Newborn Piglet

Extracorporeal membrane oxygenation (ECMO) is an important life-support system used in neonates and young children with intractable cardiorespiratory failure. In this study, we used our porcine neonatal model of venoarterial ECMO to investigate whether ECMO causes gut barrier dysfunction. We subjected 3-wk-old previously healthy piglets to venoarterial ECMO for up to 8 h and evaluated gut mucosal permeability, bacterial translocation, plasma levels of bacterial products, and ultrastructural changes in gut epithelium. We also measured plasma lipopolysaccharide (LPS) levels in a small cohort of human neonates receiving ECMO. In our porcine model, ECMO caused a rapid increase in gut mucosal permeability within the first 2 h of treatment, leading to a 6- to 10-fold rise in circulating bacterial products. These changes in barrier function were associated with cytoskeletal condensation in epithelial cells, which was explained by phosphorylation of a myosin II regulatory light chain. In support of these findings, we also detected elevated plasma LPS levels in human neonates receiving ECMO, indicating a similar loss of gut barrier function in these infants. On the basis of these data, we conclude that ECMO is an independent cause of gut barrier dysfunction and bacterial translocation may be an important contributor to ECMO-related inflammation.

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Evaluating regional blood spinal cord barrier dysfunction following spinal cord injury using longitudinal dynamic contrast-enhanced MRI

BackgroundIn vivo preclinical imaging of spinal cord injury (SCI) in rodent models provides clinically relevant information in translational research. This paper uses multimodal magnetic resonance imaging (MRI) to investigate neurovascular pathology and changes in blood spinal cord barrier (BSCB) permeability following SCI in a mouse model of SCI.MethodsC57BL/6 female mice (n = 5) were subjected to contusive injury at the thoracic T11 level and scanned on post injury days 1 and 3 using anatomical, dynamic contrast-enhanced (DCE-MRI) and diffusion tensor imaging (DTI). The injured cords were evaluated postmortem with histopathological stains specific to neurovascular changes. A computational model was implemented to map local changes in barrier function from the contrast enhancement. The area and volume of spinal cord tissue with dysfunctional barrier were determined using semi-automatic segmentation.ResultsQuantitative maps derived from the acquired DCE-MRI data depicted the degree of BSCB permeability variations in injured spinal cords. At the injury sites, the damaged barriers occupied about 70% of the total cross section and 48% of the total volume on day 1, but the corresponding measurements were reduced to 55% and 25%, respectively on day 3. These changes implied spatio-temporal remodeling of microvasculature and its architecture in injured SC. Diffusion computations included longitudinal and transverse diffusivities and fractional anisotropy index. Comparison of permeability and diffusion measurements indicated regions of injured cords with dysfunctional barriers had structural changes in the form of greater axonal loss and demyelination, as supported by histopathologic assessments.ConclusionThe results from this study collectively demonstrated the feasibility of quantitatively mapping regional BSCB dysfunction in injured cord in mouse and obtaining complementary information about its structural integrity using in vivo DCE-MRI and DTI protocols. This capability is expected to play an important role in characterizing the neurovascular changes and reorganization following SCI in longitudinal preclinical experiments, but with potential clinical implications.

Airway barrier dysfunction induced by exposure to carbon nanotubes in vitro: which role for fiber length?

Although carbon nanotubes (CNTs) are increasingly used, their biological effects are only incompletely characterized. However, experimental evidence suggests that the intratracheal instillation of CNTs causes the formation of interstitial granulomas and progressive pulmonary fibrosis in rodents. Using human epithelial Calu-3 cells as a model of airway epithelium in vitro, we have recently reported that the exposure to commercial multi-walled CNTs (MWCNTs) causes a progressive decrease of the transepithelial electrical resistance (TEER), pointing to a CNT-dependent impairment of the epithelial barrier function. To characterize better this behavior, we compared the effects of two types of MWCNTs and single-walled CNTs (SWCNTs) of different lengths on the TEER of Calu-3 monolayers. All the materials were used at a dose of 100 microg/mL corresponding to an exposure of 73 microg/cm(2) of cell monolayer. Only the longer MWCNTs and SWCNTs cause a significant decrease in TEER. To elucidate the mechanism underlying the changes in barrier function, the expression of the junction proteins occludin and ZO-1 has been also assessed. No significant decrease in the mRNA for either protein is detectable after the exposure to any type of CNTs. It is concluded that the impairment of barrier function in Calu-3 monolayers is a peculiar effect of CNTs endowed with clear cut fiber properties and is not referable to marked changes in the expression of junction proteins.

Flow‐Induced Changes in Lymphatic Endothelial Barrier Function

The purpose of this study was to determine how shear stress affects barrier function of cultured lymphatic endothelial cells. We determined the transendothelial electrical resistance (TER) of cultured human dermal microlymphatic endothelial cells (HMLEC‐d) to evaluate quantitative, real‐time changes in barrier function, during no‐flow and various flow conditions. Cells were exposed to continuous or pulsing (2 s on, 2 s off) flow at various shear stresses (2.5, 5, 9, 19, or 38 dynes/cm2) for 30 min with periods of no‐ or low‐ (0.5 dynes/cm2) flow in between. We observed a shear‐dependent increase in TER that was reversed upon turning flow off or to low at 30 min. The increase in TER with 9 dynes/cm2 was abolished by blockade of actin dynamics with 10 μM phalloidin, and significantly inhibited by blockade of Rac with 50 μM NSC23766 (9.1 ± 1.1 % increase for control vs. 2.5 ± 2.0 % with NSC23766, P<0.05). Blockade of PKA (10 μM H‐89), or ROCK (5 μM Y‐27632) did not inhibit the response. A mathematical model based on our impedance data shows that the flow‐induced changes in TER were primarily due to altered current flow between cells and not beneath cells. These results suggest that lymphatic endothelial cells dynamically alter their morphology and barrier function in response to changes in shear stress by a mechanism dependent upon Rac‐mediated actin dynamics.

Role of Ca2+/Calmodulin‐dependent Protein Kinase II (CaMKII) in Endothelial Cells

Endothelial cell (EC) barrier function and angiogenesis are partially dependent upon increases of intracellular free Ca2+ through G‐protein coupled receptor (GPCR) agonists such as thrombin and vascular endothelial growth factor (VEGF). The goal of these studies is to fully characterize the endogenous CaMKII isoform(s) in endothelial cells, and to test the hypothesis that the multifunctional Ca2+/calmodulin‐dependent serine/threonine protein kinase II (CaMKII), responds to thrombin‐ and VEGF‐stimulated Ca2+ signals and regulates EC barrier function and angiogenesis. For the 1st time, by RT‐PCR and western blotting, we successfully characterized the predominant CaMKII isoform expressed in cultured human umbilical vein endothelial cells (HUVEC) is the delta 6 (δ6) isoform, a novel δ‐isoform lacking the 21aa c‐terminal domain. We utilized siRNA approach to suppress CaMKIIδ6 protein expression and found it attenuated, only low dose (2.5nM) but not high dose (10nM), of thrombin‐induced increase of EC permeability. These data suggest a role of CaMKIIδ6 in mediating changes in barrier function in response to physiological relevant doses of thrombin. By using the same approach, we found that CaMKIIδ6 is pivotal in regulating VEGF induced HUVEC migration through Boyden Chamber. This work was supported by NIH grant to H.A.S (R01 HL‐049426).Grant Funding SourceNIH grant to H.A.S (R01 HL‐049426).

Differential Regulation of Conjunctival Epithelial Permeability by Staphylococcus Enterotoxin B, Platelet Activating Factor, and IL-8

RATIONALE: Ocular exposure to Staphylococcal enterotoxin B (SEB) can result in systemic effects, yet it is not known whether exposure of conjunctival epithelium to SEB results in changes in barrier function which promote access of SEB to underlying tissues. The purpose of this study was to examine regulation of human conjunctival epithelial (HCE) permeability in response to SEB in comparison to endogenous stimuli (PAF, IL-8) known to promote changes in permeability in other epithelial tissues.

CD24 regulated gene expression and distribution of tight junction proteins is associated with altered barrier function in oral epithelial monolayers

BackgroundControl of intercellular penetration of microbial products is critical for the barrier function of oral epithelia. We demonstrated that CD24 is selectively and strongly expressed in the cells of the epithelial attachment to the tooth and the epithelial lining of the diseased periodontal pocket and studies in vitro showed that CD24 regulated expression of the epithelial intercellular adhesion protein E-cadherin.ResultsIn the present study, the barrier function of oral epithelial cell monolayers to low molecular weight dextran was assayed as a model for the normal physiological function of the epithelial attachment to limit ingress of microbial products from oral microbial biofilms. Paracellular transfer of low molecular weight dextran across monolayers of oral epithelial cells was specifically decreased following incubation with anti-CD24 peptide antibody whereas passage of dextran across the monolayer was increased following silencing of mRNA for CD24. Changes in barrier function were related to the selective regulation of the genes encoding zonula occludens-1, zonula occludens-2 and occludin, proteins implicated in tight junctions. More particularly, enhanced barrier function was related to relocation of these proteins to the cell periphery, compatible with tight junctions.ConclusionCD24 has the constitutive function of maintaining expression of selected genes encoding tight junction components associated with a marginal barrier function of epithelial monolayers. Activation by binding of an external ligand to CD24 enhances this expression but is also effective in re-deployment of tight junction proteins that is aligned with enhanced intercellular barrier function. These results establish the potential of CD24 to act as a potent regulator of the intercellular barrier function of epithelia in response to local microbial ecology.

Dose-Dependent Effects of Ethanol and E. Coli on Gut Permeability and Cytokine Production

The gut may prime inflammatory responses following shock/trauma insults. Ethanol (EtOH) use is common in trauma patients and may impair intestinal barrier function. We compared varying concentrations of EtOH on proinflammatory cytokine production of Caco2 cell monolayers and the resultant changes in barrier function. We hypothesized that even low concentrations of EtOH would cause significant cytokine release and barrier dysfunction in vitro. Confluent Caco2 cell monolayers were grown in a two-chamber culture system and exposed to varying concentrations of EtOH (0.1%, 0.5%, 1.0%, 1.5%, and 2.0%) with/without Escherichia coli C-25 (EC). Supernatants were collected and TNF and IL6 quantified by ELISA (pg/mL). Monolayer integrity was assessed by apoptosis and permeability measurements. Caco2 production of TNF-alpha increased in a dose-dependent manner when incubated with increasing concentrations of EtoH. A synergistic effect was seen when E. coli was added to the apical chamber. A similar result was seen with the production of IL-6. A dose-dependent effect was also noted with EtOH with or without E. coli on apoptosis and permeability measurements. In addition to alterations in gut permeability, increasing concentrations of ethanol have a synergistic effect with E. coli on Caco2 production of proinflammatory cytokines TNF and IL-6. The creation of a proinflammatory cytokine milieu with an altered barrier integrity may be a mechanism by which ethanol may increase septic complications in the injured patient.

Atopic dermatitis: Therapeutic concepts evolving from new pathophysiologic insights

Recent insights into the relevance of the epidermal barrier function and its interaction with components of the innate and adaptive immune responses in patients with atopic dermatitis (AD) give rise to a number of novel potential treatment options. In particular, the identification of loss-of-function mutations in the barrier protein filaggrin and of a diminished expression of certain antimicrobial peptides in AD skin stimulates new concepts to think beyond the T(H)1/T(H)2 paradigm. This review will focus on these most recent discoveries and will discuss new and corresponding proof-of-concept trials in patients with AD. It will further speculate on novel ways to restore the homeostasis among the 3 major components in AD skin suspected to be clinically relevant.

Impaired glycocalyx barrier properties and increased capillary tube haematocrit

Impaired glycocalyx barrier properties and increased capillary tube haematocrit

Barrier functions and paracellular integrity in human cell culture models of the proximal respiratory unit

Airway epithelial cells provide a barrier to the translocation of inhaled materials. Tight (TJ) and adherens junctions (AJ) play a key role in maintaining barrier functions, and are responsible for the selective transport of various substances through the paracellular pathway. In this study we compared a bronchial cell line (16HBE14o-) and primary bronchial cells (HBEC), both cocultivated with the fibroblast cell line Wi-38, with respect to their structural differentiation and their reaction to cytokine stimulation. HBEC formed a pseudostratified epithelial layer and expressed TJ and AJ proteins after 2 weeks in coculture. Mucus-producing and ciliated cells were found within 24 days. Additionally, a beating activity of the ciliated HBEC (14–19 Hz) could be detected. 16HBE14o- in coculture showed a multilayered growth without differentiation to a pseudostratified airway epithelium. Simultaneous exposure to TNF-α- and IFN-γ-induced significant changes in barrier function and paracellular permeability in the cocultures of HBEC/Wi-38 but not in the 16HBE14o-/Wi-38. In summary, HBEC in coculture mimic the structure of native polarized bronchial epithelium showing basal, mucus-producing and ciliated cells. Our system provides an opportunity to examine the factors that influence barrier and mucociliary function of bronchial epithelium within a time frame of 3 weeks up to 3 months in an in vivo-like differentiated model.

A randomized investigator‐blind trial of different passes of microdermabrasion therapy and their effects on skin biophysical characteristics

Microdermabrasion (MDA) is a safe, simple, and beneficial technique for superficial skin resurfacing. Despite its popular usage, few studies have assessed the efficacy of different MDA protocols applied at the present time. Objectives To assess the effects of MDA generally, as well as to compare the effects of two vs. three passes of MDA in each session for a total number of six therapeutic sessions on skin biophysical characteristics. In this randomized, investigator-blind, split-face study, 10 patients underwent a series of six MDA treatments with an interval of 2 weeks. One side of the face was treated with two passes of MDA and the other side was treated with three passes, randomly. Stratum corneum hydration, sebum secretion, and skin pH measurements were obtained before and after the procedure on all sessions and also 1 and 4 weeks after the last treatment. After six sessions of MDA, a decrease in sebum content compared to baseline was shown at the end of treatment sessions, but no statistical difference was observed between two vs. three passes groups (-30.0 [interquartile range, IQR = 50.0] vs. -27.5 [IQR = 125.3], respectively, P = 0.58). Comparison of two treatment groups showed significant higher values of sebum content in the first follow-up after treatment with three passes of MDA. (64.0 [IQR = 52.0] for three passes vs. 45.0 [IQR = 46.0] for two passes, P = 0.04) A significant increase was observed in pH values at the end of treatment series, first and second follow-up after treatment with two passes of MDA. MDA may have remarkable effects on skin barrier function changes resulting in skin clinical improvements (Cochrane Skin Group identifier: CSG No. 37).

Change of biophysical properties of the skin caused by ultraviolet radiation‐induced photodamage in Koreans

Ultraviolet (UV) irradiation affects the function and complexion of the skin by inducing changes in physical properties through formation of erythema, proliferation of epithelial cells, DNA damage, activation or inactivation of various enzymes and proteins, and free radical formation. In this study, the authors intended to observe the overall course of changes in barrier function and reflectance of the skin induced by photodamage, and healing reaction in the course of time, and alteration of the skin complexion. The subjects were chosen from 15 healthy Korean men 20-35 in age, that fall into the category of Fitzpatrick's skin types II, III, and IV without history of recent exposure to sunlight, photosensitivity, or having taken any drugs that induce phototoxicity or photoallergic reactions. The subjects were artificially exposed to suberythemogenic dose [0.5 minimal erythemal dose (MED), 0.75 MED], 1 MED and high dose (2.5 MED) by solar simulator, and changes in skin barrier function and skin reflectance were assessed with a Tewameter, a Corneometer, and a Colorimeter for 4 weeks. Transepidermal water loss (TEWL) increased abruptly at Day 1 of single solar UV (SSUV) exposure, and slowly returned to the original level from Day 2/Day 3. In the case of exposure with 0.75 MED, it returned to the original level at Day 4 of exposure, and at Days 7 and 28 in the cases of 1 MED and 2.5 MED exposure, respectively. Water-holding capacity sharply declined at Day 1 of exposure, hitting the lowest point at Day 2, and then slowly recovered starting on Day 3. In the case of exposure with 0.75 MED and 1 MED, it returned to the original level at Days 7 and 28 in the case of 2.5 MED exposure. The a(*) values abruptly increased and reached the peak at Day 1 and slowly returned to the original level at Day 2, while the b(*) values slowly increased at Day 3, peaking at Day 7 and slowly returning to the original level thereafter. The L(*) values abruptly declined at Day 1, maintaining plateau through Day 7 and slowly returning to the baseline level thereafter. The individual typology angle (ITA degrees ) were compatible with L(*) values change. The erythema index increased abruptly at Day 1 of SSUV exposure, peaking at Day 2 and slowly returned to the original level starting at Day 3. Melanin index slowly started to increase on Day 3 of SSUV exposure, peaking at Day 7 and gradually returned to the original level thereafter. However, L(*), a(*), b(*), erythema index, and melanin index did not return to the original level during the 28-day course of this study. This study shows that in the skin of Korean subjects, changes in skin barrier function and delayed melanization do occur even in exposure to a suberythemogenic dose of SSUV. Also, given the fact that restoration of barrier function occurs as the process of melanization begins, melanization is considered to be a useful predictive indicator of the restoration of the skin barrier function after sunburn.

Induction of claudins in passaged hTERT-transfected human nasal epithelial cells with an extended life span

The epithelial barrier of the upper respiratory tract, such as that of the nasal mucosa, plays a crucial role in host defense. The epithelial barrier is regulated in large part by the apical-most intercellular junctions, referred to as tight junctions. However, the mechanisms regulating of tight junction barrier in human nasal epithelial cells remain unclear because the proliferation and storage of epithelial cells in primary cultures are limited. In the present study, we introduced the catalytic component of telomerase, the hTERT gene, into primary cultured human nasal epithelial cells and examined the properties of the transfectants, including their expression of tight junctions, compared with primary cultures. The ectopic expression of hTERT in the epithelial cells resulted in adequate growth potential and a longer lifespan of the cells. The properties of the passaged hTERT-transfected cells including tight junctions were similar to those of the cells in primary cultures. The barrier function in the transfectants after treatment with 10% FBS was significantly enhanced with increases of integral tight junction proteins claudin-1 and -4. When the transfectants were treated with TGF-beta, which is assosciated with nasal polyposis and chronic rhinosinusitis, upregulation of only claudin-4 was observed, without a change of barrier function. In human nasal epithelial cells, the claudins may be important for barrier function and a novel target for a drug-delivery system. Our results indicate that hTERT-transfected human nasal epithelial cells with an extended lifespan can be used as an indispensable and stable model for studying the regulation of claudins in human nasal epithelium.