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PubMed2000

Evaluation of dentin root canal permeability after instrumentation and Er:YAG laser application.

BACKGROUND AND OBJECTIVES: Smear layer removal with EDTA from root canal walls allows greater cleaning and disinfection of root canals. However, because Er:YAG laser acts on the removal of the smear layer, the objective of investigation was to analyze in vitro the effect of Er:YAG laser on dentin root canal wall permeability after endodontic instrumentation and irrigation with water or sodium hypochlorite and Er:YAG laser application. STUDY DESIGN/MATERIALS AND METHODS: A total of 25 extracted human maxillary incisors were divided into five groups: Group I, instrumentation with deionized distilled water as the irrigating solution; Group II, instrumentation with 1% sodium hypochlorite as the irrigating solution; Group III, instrumentation with deionized distilled water as the irrigating solution and Er:YAG laser application; Group IV, instrumentation with 1% sodium hypochlorite solution as the irrigating solution and Er:YAG laser application; Group V, instrumentation only up to #20 file with deionized distilled water as the irrigating solution and Er:YAG laser irradiation. The laser parameters were 15 Hz, 140 mJ, total energy 42 J, 300 pulses (Kavo Key Laser). Copper sulfate (10%) was used to evaluate dentin permeability. The penetration of copper ions into the dentinal tubules was observed using 1% rubeanic acid, which reveals copper ions, forming a stained compound ranging in color from deep blue to black. Transverse sections (500-microm thick) were obtained with a diamond disk from the cervical, middle, and apical thirds. RESULTS: The instrumentation of the root canal that used water as the irrigating solution followed by Er:YAG laser irradiation promoted the greatest increase in dentin permeability. The use of Er:YAG laser, 1% sodium hypochlorite + Er:YAG, and 1% sodium hypochlorite used alone showed an intermediate capacity of increasing dentin permeability. The use of water as the irrigating solution without Er:YAG laser promoted the least dentin permeability. CONCLUSIONS: The use of water as the irrigating solution after instrumentation and Er:YAG laser irradiation was an effective procedure for increasing dentin permeability.

Dental Pulp Cavity↗
PubMedDate not supplied

Structure and permeability of human placental microvasculature.

Endothelial paracellular junctions are important structures for the regulation of vascular permeability, junctional organisation being systematically related to the functional properties of the endothelium. Electron microscopic studies, immunocytochemistry, and single-passage permeability measurements have established that the placental microvessels resemble the fairly tight continuous microvessels of skeletal muscle both in structure and permeability. The endothelial paracellular clefts of these microvessels contain two distinct junctional entities which may influence permeability: the tight junction and the adherens junction. These clefts impose a substantial restriction to molecules above RMM 1000 Da and large haemproteins cannot cross the clefts. The 18 nm-wide zones of the clefts possess the transmembrane adhesion molecules PECAM-1 and VE-cadherin, which have been implicated in junctional assembly and permeability. Inflammatory mediators such as histamine and tumour necrosis factor cause a redistribution of these adhesion molecules to non-junctional regions, and histamine (100 microM) causes a rapid and sustained rise in extraction of radio-labeled tracers. Electron microscopy has also revealed possible first indications of tight junctional disassembly. Both the endothelia of larger placental vessels and isolated placental microvascular endothelial cells express cadherins and PECAM-1 and contain an extensive F-actin cytoskeleton, which is implicated in changes of cell shape and junctional assembly/disassembly. Thus, the human placental endothelium, using perfusion techniques and in vitro experiments, offers a valuable model for vascular permeability studies.

Female↗
PubMed1997

Water distribution and permeability of zebrafish embryos, Brachydanio rerio.

Teleost embryos have not been successfully cryopreserved. To formulate successful cryopreservation protocols, the distribution and cellular permeability to water must be understood. In this paper, the zebrafish (Brachydanio rerio) was used as a model for basic studies of the distribution to permeability to water. These embryos are a complex multi-compartmental system composed of two membrane-limited compartments, a large yolk (surrounded by the yolk syncytial layer) and differentiating blastoderm cells (each surrounded by a plasma membrane). Due to the complexity of this system, a variety of techniques, including magnetic resonance microscopy and electron spin resonance, was used to measure the water in these compartments. Cellular water was distributed unequally in each compartment. At the 6-somite stage, the percent water (V/V) was distributed as follows: total in embryo = 74%, total in yolk = 42%, and total in blastoderm = 82%. A one-compartment model was used to analyze kinetic, osmotic shrinkage data and determine a phenomenological water permeability parameter, Lp, assuming intracellular isosmotic compartments of either 40 or 300 mosm. This analysis revealed that the membrane permeability changed (P < 0.05) during development. During the 75% epiboly to 3-somite stage, the mean membrane permeability remained constant (Lp = 0.022 +/- 0.002 micron x min-1atm-1 [mean +/- S.E.M.] assuming isosmotic is 40 mosm or Lp = 0.049 +/- 0.008 micron x min-1atm-1 assuming isosmotic is 300 mosm). However, at the 6-somite stage, Lp increased twofold (Lp = 0.040 +/- 0.004 micron x min-1atm-1 assuming isosmotic is 40 mosm or Lp = 0.100 +/- 0.017 micron x min-1atm-1 assuming isosmotic is 300 mosm). Therefore, the low permeability of the zebrafish embryo coupled with its large size (and consequent low area to volume ratio) led to a very slow osmotic response that should be considered before formulating cryopreservation protocols.

Animals↗
PubMed1996

Flow-induced modulation of the permeability of endothelial cells cultured on microcarrier beads.

The maintenance of endothelial barrier function is important in the regulation of fluid and solute balance between the vascular space and the surrounding tissue. Since fluid flow across endothelial cells stimulates a wide variety of endothelial responses, the effect of shear stress on barrier function was investigated. Bovine pulmonary artery endothelial cells were cultured on permeable microcarrier beads, placed in a chromatography column, and perfused. Indicator-dilution techniques were used to estimate the permeability of the cell-covered beads to low molecular weight tracers (sodium fluorescein-NaFlsc; cyanocobalamin-B12) as a function of flow rate through the column. Permeability values for both tracers were significantly increased (9.3 +/- 0.6 to 19.3 +/- 1.7 for NaFlsc; 8.2 +/- 0.5 to 20.4 +/- 3.1 for B12; mean+/-SEM, x 10(-5) cm/s, P < .05) when the flow rate was increased from 0.9 ml/min to 3.2 ml/min (corresponding to average shear stresses of 4.7 and 16.8 dynes/cm2). The permeability increase occurred within minutes of the flow increase, and was reversed by decreasing the flow rate to 0.9 ml/min. In the presence of cytochalasin D, the flow-induced permeability increase was not reversible. Neither inhibition of nitric oxide synthase (with NG-monomethyl-L-arginine) nor inhibition of cyclooxygenase (with indomethacin) was capable of blocking the flow-induced permeability increase. These results indicate that the rapid modulation of endothelial barrier by flow in vitro is probably not due to prostacyclin or nitric oxide.

Animals↗
PubMedDate not supplied

Increasing endothelial cell permeability improves the efficiency of myocyte adenoviral vector infection.

BACKGROUND: Gene delivery to the myocardium using blood-borne adenoviral vectors is hindered by the endothelium, which represents a barrier limiting the infection rate of underlying myocytes. However, endothelial permeability may be modulated by pharmacological agents. METHODS: In the present study, we modeled the endothelial barrier in vitro using a human umbilical vein endothelial cell (HUVEC) monolayer seeded on a Transwell membrane as a support and diffusion of fluorescent dextrans as a permeability index. We used alpha-thrombin (100 nM) as a pharmacological agent known to increase endothelial permeability and tested the barrier function of the endothelial cell monolayer on adenovector-mediated luciferase gene transfer to underlying isolated cardiac myocytes. RESULTS: A confluent HUVEC monolayer represented a considerable physical barrier to dextran diffusion; it reduced the permeability of the micropore membrane alone to fluorescein isothiocyanate (FITC)-labeled dextrans of molecular weights 4, 70, 150 and 2000 kDa by approximately 54, 78, 88 and 98%, respectively. Alpha-thrombin (100 nM) increased the permeability coefficients (P(EC)) by 276, 264, 562 and 4166% for the same dextrans, respectively. A confluent HUVEC monolayer represented a major impediment to adenovector-mediated luciferase gene transfer to cardiac myocytes, largely reducing gene transfer efficiency. However thrombin induced a nine-fold increase in myocyte infection. CONCLUSION: In our model, the endothelial cell monolayer represents a major impediment to myocyte adenovector-mediated gene transfer which can be partially improved by pharmacologically increasing endothelial permeability. The Transwell model is therefore particularly useful for testing the efficiency of pharmacological agents in modulating adenovector passage through the endothelial barrier.

Adenoviridae↗
PubMed1980

Permeability of hyaluronic acid solutions. The effects of matrix concentration, calcium, and pH.

The permeability of hyaluronic acid solutions was measured in a chamber closed at each end of semi-permeable membranes. Permeability was strongly concentration dependent below 1.0 gm% hyaluronic acid. Small changes in the ionic environment of 1.0 gm% hyaluronic acid solutions also altered permeability. The permeability of such solutions decreased by a factor of 4.7 in the presence of as little as 5 mM calcium ion and showed optimum permeability at pH 7.0. The pH optimum was not noted in hyaluronic acid preparations known to contain only small amounts of protein.

Calcium↗
PubMed1981

Permeability of rheumatoid and normal human synovium to specific plasma proteins.

A method is described for the determination of the permeability of the blood-joint barrier to specific plasma proteins, using the ratio of protein concentration in synovial fluid to that in plasma. The inadequacy of the ratio per se as a direct index of permeability is discussed. Permeabilities are evaluated for the normal and rheumatoid human knee. Permeability increases in the rhematoid knee by approximately 6 times for the rheumatoid knee by approximately 6 times for albumin and over 40 times for macroglobulins. The effect of protein molecular dimensions upon permeability is analyzed. Permeability shows less dependence upon solute dimensions in the rheumatoid knee than in the normal knee, i.e., molecular selectivity is reduced. From these data and synovial morphology, a two-membrane model of the blood-joint barrier is developed. The relative contribution of the component intimal and endothelial layers to the total barrier is found to depend upon solute dimensions.

Arthritis, Rheumatoid↗
PubMed1991

Nerve cables formed in silicone chambers reconstitute a perineurial but not a vascular endoneurial permeability barrier.

The passage of molecules into the endoneurial environment of the axons of normal peripheral nerve is regulated by two permeability barriers, the perineurial-nerve barrier and the endoneurial blood-nerve barrier. These barriers exist because of the presence of tight junctions between adjacent perineurial cells and adjacent endothelial cells. In the present study we investigated whether permeability barriers form in nerve cables, which develop inside silicone chambers. The sciatic nerves of adult rats were cut, and the proximal and distal ends sutured into opposite ends of silicone chambers that were filled with dialyzed plasma. The presence of barriers was determined with the tracer horseradish peroxidase (HRP), which was injected intravenously and detected histochemically in tissues by light and electron microscopy. At four weeks, a regenerated nerve cable extended across the 10 mm length of each chamber. However, no permeability barriers were present since the reaction product for HRP was visible throughout the cable. At twenty-six weeks, all the axons in cables were gathered into minifascicles. Each minifascicle of axons was surrounded by perineurial cells. Blood vessels were excluded from the minifascicles by the perineurial cells and the vessels were permeable to HRP, thus indicating that their endothelial cells had not formed tight junctions. Despite the leakage of HRP from the excluded vessels, the tracer did not reach the axons because the perineurial cells encircling the minifascicles developed tight junctions. In some animals, the chambers were removed at four weeks to determine whether the chamber influenced barrier development. This manipulation had no effect since cables, with or without chambers, exhibited similar findings at twenty-six weeks. Our results indicate that nerve cables regenerate a perineurial but not an endoneurial permeability barrier. We conclude that axons in long-term cables are protected by only a perineurial permeability barrier.

Animals↗
PubMedDate not supplied

Permeability characteristics of the guinea pig biliary apparatus.

To determine the mechanism and pathway of entry of polar nonelectrolytes into bile, we studied first the permeation of [3H]H2O, [14C]urea, [14C]erythritol, [14C]mannitol, [3H]sucrose, [3H]inulin and [3H]dextran across an isolated, in situ perfused segment of the guinea pig's extrahepatic bile duct. All of these molecules, except [3H]dextran, permeated the bile duct. The diffusive permeability coefficients (cm per sec per 10(6) ranged from 248 for [3H]H2O to 1.2 for [3H]inulin. On the basis of these results, we formulated two models of solute biliary excretion. In Model I, permeation across both the canaliculus and ductule/duct was assumed to be by simple diffusion. In Model II, solute was assumed to enter the canaliculus by convection and diffusion, and the ductule/duct by diffusion. Reflection coefficients and/or permeability coefficients for the canalicular membrane were then determined by fitting the equations describing these processes to the bile-to-plasma ratios of [14C] erythritol, [14C]mannitol, [3H]sucrose and [3H]inulin observed at different rates of bile flow produced by partially clamping the bile duct cannula and subsequently infusing taurodehydrocholate. Only when convection was included as a mechanism of canalicular permeation (Model II) was excellent fitting observed. In such a case, the reflection coefficients and permeability coefficients for the carbohydrates in question were similar to those reported for other transporting epithelia. Cholestasis produced by taurolithocholate (10 to 40 mumoles per kg, i.v.) was associated with an irreversible increase in both the sieving coefficient and permeability coefficient for [3H]sucrose and [3H]inulin, even when the inhibition of bile flow was fully reversible. The permeability to [14C] erythritol and [14C]mannitol was either not affected or minimally increased. These findings suggest that, in the guinea pig: solutes as large as [3H]inulin enter the biliary tree both at the canaliculus and bile ductule/duct; [14C]erythritol and [14C]mannitol gain access into the canalicular lumen primarily by convection, whereas [3H]sucrose and [3H]inulin permeate mainly by diffusion; distal permeation of these carbohydrates is small, and accounts for 1 to 11% of their total biliary entry; the canalicular membrane permeability to [3H]sucrose and [3H]inulin is not functionally important in bile secretion, and a fraction of canalicular bile flows through the transjunctional shunt pathway.

Animals↗
PubMed1994

Mathematical modeling of water permeability of surgical fabrics for vascular use.

The purpose of this study was to derive new mathemtic formulae that could be used reliably to predict water permeability of surgical fabrics before they are made and tested for water permeability. Such a theoretical prediction of water permeability, Qprd, of surgical fabrics is needed for not only timely characterization but also for assisting in more efficient future design and development of better surgical fabrics. Two mathematic formulae, Qw and Qk, were derived from the Buckingham Pi Theorem, in which relevant fiber and fabric parameters were placed into dimensionless pi groups and computed for 25 commercial and experimental vascular fabrics. Linear regression analysis of the relationship between these pi groups with water permeability on the woven and knitted grafts yielded coefficients for the corresponding pi groups that were required for constructing appropriate mathematic formulae to predict water permeability of vascular fabrics. When proper sources of the experimentally determined water permeability, Qexp, were chosen for comparison, we found that 86% of woven fabrics (6 of 7) and 77% of knitted fabrics (14 of 18) had their Qprd within 10% of their Qexp. This high percentage of close matching (within 10%) between Qprd and Qexp should be considered satisfactory because the experimental error for obtaining Qexp is generally higher than 10%. The difference between Qprd and Qexp ranged from as small as 0.27% to as high as 74.2%, depending on the type of fabrics and source of Qexp.(ABSTRACT TRUNCATED AT 250 WORDS)

Models, Theoretical↗
PubMed2002

Platelet-derived growth factor mediates tight junction redistribution and increases permeability in MDCK cells.

Increased tissue permeability is a common characteristic of a number of diseases such as pulmonary edema, inflammatory bowel disease, several kidney diseases, diabetic retinopathy, and tumors. We hypothesized that growth factors increase permeability by redistribution of tight junction proteins away from the cell border. To investigate mechanisms of growth factor-mediated permeability, we examined the effect of platelet derived growth factor (PDGF) on Madin-Darby canine kidney (MDCK) cell tight junction protein distribution and on permeability. PDGF altered the cellular distribution of occludin and ZO-1 from the cell border to the cytoplasm and increased permeability to 70 kDa dextran in a concentration-dependent manner. Treatment of MDCK cells with PDGF prior to fixation allowed binding of the lectin concanavalin A to the basement membrane of fixed cells, while binding was prevented in untreated control monolayers, implying that PDGF induced the formation of a paracellular transport pathway. Cell fractionation experiments with PDGF-treated cells revealed a novel occludin-containing low-density, detergent resistant subcellular structure, which increased in the buoyant fractions relative to occludin in the pellet in a time- and concentration-dependent manner. Immunocytochemistry revealed that a pool of internalized occludin co-labels with the early endosome marker, EEA1, suggesting that PDGF may stimulate occludin to enter an endosomal pathway. PDGF may act as a permeabilizing agent by moving tight junction proteins away from the cell border in discrete microdomains, and the effects of PDGF on permeability and tight junction protein distribution may model the regulation of epithelial and endothelial barrier properties by other peptide growth factors.

Animals↗
PubMed1988

Effects of human neutrophil chemotaxis across human endothelial cell monolayers on the permeability of these monolayers to ions and macromolecules.

We have developed a method for studying the permeability properties of human endothelia in vitro. Human umbilical vein endothelial cells (HUVEC) were cultured on a substrate of human amnion. Confluent monolayers of these cells demonstrated 6-12 delta.cm2 of electrical resistance (a measure of their permeability to ions) and restricted the transendothelial passage of albumin from their apical to their basal surface. To determine whether leukocyte emigration alters endothelial permeability in this model, we examined the effects of migrating human polymorphonuclear leukocytes (PMN) on these two parameters. Few PMN migrated across the HUVEC monolayers in the absence of chemoattractants. In response to chemoattractants, PMN migration through HUVEC monolayers was virtually complete within 10 minutes and occurred at random locations throughout the monolayer. PMN migrated across the monolayer via the paracellular pathway. Although one PMN migrated across the monolayer for each HUVEC, PMN migration induced no change in electrical resistance or albumin permeability of these monolayers. At this PMN:HUVEC ratio, these permeability findings were correlated morphologically to measurements that HUVEC paracellular pathway size increases by less than 0.22% with PMN migration. This increase is insufficient to effect a measurable change in the electrical resistance of the endothelial cell monolayer. These findings demonstrate that increased permeability of cultured endothelial cell monolayers is not a necessary consequence of PMN emigration.

Albumins↗
PubMed1990

Tumor necrosis factor enhances the neutrophil-dependent increase in endothelial permeability.

We examined the effect of tumor necrosis factor alpha (TNF alpha) on the increase in pulmonary microvascular endothelial monolayer permeability induced by activated neutrophils (PMN). Layering of PMN onto endothelial monolayers followed by activation of PMN with phorbol 12-myristate 13-acetate (PMA) increased 125I-albumin clearance rate across the monolayers. Pretreatment of endothelial monolayers for 6 hr with TNF alpha (200 U/ml) potentiated the PMN-dependent increase in endothelial permeability, whereas 1 hr or 6 hr pretreatment of endothelial monolayers with 200 U/ml and 100 U/ml, respectively, TNF alpha did not enhance the response. Adherence of PMN to the endothelial cells was increased at 1 and 6 hr after TNF alpha (200 U/ml) treatment, but the adherence response was markedly greater following 6 hr of TNF alpha. The TNF alpha treatment of endothelial cells did not enhance neutrophil activation responses to PMA. Pretreatment of PMN with IB4, a MAb to the CD18 integrin, the common beta subunit of the adhesion proteins LFA-1, Mac-1, and p150,95 of PMN, reduced the increases in PMN adherence and the endothelial monolayer permeability induced by the 6 hr TNF alpha treatment. In contrast, pretreatment of PMN with OKM-1, a MAb to the CD11b epitope (alpha-subunit), had no effect on the adherence and the potentiation of the increase in permeability. The potentiation of the PMN-dependent permeability increase and enhanced endothelial adhesivity at 6 hr after TNF alpha priming of endothelial cells was dependent on protein synthesis. The results indicate that protein synthesis-dependent expression of an endothelial ligand for CD18 and resultant endothelial hyperadhesiveness potentiates the PMN-mediated increase in endothelial permeability after TNF alpha activation of endothelial cells. The priming of endothelial cells by TNF alpha may be a critical step in the mediation of endothelial injury.

Animals↗
PubMed1992

Effects of acute vs. chronic phorbol ester exposure on transepithelial permeability and epithelial morphology.

In previous experiments we have shown that acute (30 minutes) exposure to phorbol esters or other protein kinase C activators causes increased transepithelial permeability, specifically by the increased paracellular permeability through tight junctions. However, the role of protein kinase C activators in carcinogenesis is predicted upon a chronic exposure of an effective dose at frequent intervals for a prolonged period of time. We therefore sought to determine the effect of chronic phorbol ester exposure on transepithelial permeability by exposing cells of the polar renal epithelial cell line, LLC-PK1, to phorbol esters for time periods as long as 16 weeks. The following changes ensued: (1) after the initial drop in transepithelial resistance due to phorbol ester exposure, i.e., an increase in transepithelial permeability (in the acute phase of exposure), an adaptive response occurs as transepithelial resistances in chronically exposed cultures recover to approximately 50% of control values, (2) the cell sheets in chronically exposed cultures lose their acute responsiveness of transepithelial permeability to phorbol ester exposure, (3) cell sheet architecture changes as cells occasionally multilayer and actual polyp-like cell masses appear at high frequency, and (4) cytosolic protein kinase C activity decreases to 50% of control level with acute exposure and then is further decreased to less than 1% of control level in chronically treated cells; membrane-associated PKC activity is not as sharply decreased. The possible role of transepithelial permeability in carcinogenesis and the value of chronically treated epithelial cell cultures as a model for two-stage carcinogenesis are discussed.

Animals↗
PubMed1984

Regulation of the permeability of the medaka fish embryo chorion by exogeneous sodium and calcium ions.

We questioned if the optically transparent noncellular chorion, or egg envelope, which encapsulates the entire medaka fish (Oryzias latipes) embryo might in some way constitute a permeability barrier to high concentrations of the diuretic called amiloride. More specifically, we questioned if removal of cations from the exogenous environment of the medaka embryo might make the chorion more permeable to amiloride and thereby make the fish embryos more sensitive to the inhibitory and lethal effects of this drug. To test this question, chorion-encapsulated medaka embryos were exposed to: deionized-distilled water, to Yamamoto-Ringer's (Y-R) solution, to Yamamoto-Ringer's containing choline chloride as a substitute for NaCl, and to isotonic NaCl solution in the presence of and in the absence of amiloride. Briefly, the prediction that the medaka embryos would be most sensitive to amiloride's inhibitory effects in distilled water was confirmed. Further studies showed that the presence of Na+ or of Ca2+ alone in the culture solution gave partial protection against the lethal effects of the amiloride. Electron probe X-ray microanalysis studies indicated that addition of Ca2+ and other cations to the culture solution caused the concentrations of cations to increase in the chorion, and that increase was correlated to a visible decrease in the permeability of the chorion to the amiloride. This decreased permeability of the chorion apparently protected the embryo from the amiloride. The decreased permeability of the chorion to amiloride, which occurred in the presence of the cations present in Y-R solution, was found to be reversible once the cations were washed from the chorion. Key words medaka, chorion, Na+, Ca2+ permeability, x-ray microanalysis, Oryzias latipes, egg envelope.

Amiloride↗
PubMed2004

Assessment of the effects of sodium dodecyl sulfate on the buccal permeability of caffeine and estradiol.

The concentration-dependent effects of sodium dodecyl sulfate (SDS) on the in vitro permeability of the buccal mucosa were assessed using caffeine (CAF) and estradiol (E(2)) as model hydrophilic and lipophilic markers, respectively. The permeability of CAF and E(2) through porcine buccal mucosa was determined in modified Ussing chambers, with and without exposure to different concentrations of SDS (0.01, 0.05, 0.1, and 1% w/v in physiological buffer). Permeability experiments were complemented with light microscopic evaluation of untreated and SDS-treated tissues. Additionally, the critical micellar concentration of SDS in the physiological buffer and the effect of SDS pretreatment on drug solubility were determined. Pretreatment of buccal mucosa with SDS 0.01% had no effect on CAF or E(2) permeability. SDS 0.05, 0.1, and 1% significantly enhanced CAF flux by a factor of 1.57, 1.63, and 1.81, respectively, and caused significant removal of superficial cells, as observed with light microscopy. Interestingly, pretreatment with SDS 0.05% did not affect E(2) flux, whereas SDS at > or =0.1% significantly reduced E(2) permeability, possibly as a result of micellar solubilization. These results demonstrate that the effect of SDS on buccal permeability depends on both the concentration of SDS used and the physicochemical properties of the permeant.

Animals↗
PubMed2004

Experimental demonstration of the existence of highly permeable localized transport regions in low-frequency sonophoresis.

Recent advances in low-frequency sonophoresis have focused on the existence of hypothesized localized transport regions (LTRs). However, there has been no actual experimental demonstration that the hypothesized LTRs are, in fact, localized regions of high permeability. Through a series of low-frequency sonophoresis experiments conducted with full-thickness pig skin, in the presence of the surfactant sodium lauryl sulfate (SLS), in which we have separately measured the transport of calcein through the LTRs, which have areas ranging from 10 to 40 mm(2), and the surrounding regions of the skin (the non-LTRs) by means of a novel masking technique, we demonstrate that the calcein permeability through the LTRs is approximately 80-fold higher than the calcein permeability through the non-LTRs, suggesting that the LTRs are structurally perturbed to a greater extent than the non-LTRs from the exposure to the ultrasound/SLS system. In addition, we propose basic models to predict the total skin transdermal permeability from the transdermal permeabilities of the LTRs and the non-LTRs, and then compare the predictions to the experimental data obtained from the masking experiments. We also demonstrate that both the LTRs and the non-LTRs exhibit significant decreases in skin electrical resistivity relative to untreated skin ( approximately 5000-fold and approximately 170-fold, respectively), suggesting the existence of two levels of significant skin structural perturbation due to ultrasound exposure in the presence of SLS. Finally, an analysis of the porosity/tortuosity ratio (epsilon/tau) values suggests that trans-cellular transdermal transport pathways are present within the highly permeable, and highly structurally perturbed, LTRs.

Animals↗
PubMed1990

Effect of adjuvants on charge-selective permeability and electrical resistance of rat jejunal membrane.

Effects of disodium ethylenediaminetetraacetate (EDTA) and bile salts on the ion-selective permeability and electrical resistance (Rm) of rat jejunal membrane were investigated in vitro to characterize the enhanced permeability to drugs by these adjuvants. Ion-selective permeability was estimated from diffusion potentials across the membrane and expressed as permeability ratios for ions (Pcl/PNa, PK/PNa). Pretreatment of the mucosal membrane with EDTA increased PCl/PNa and decreased Rm. At the same time, the mucosal-to-serosal flux rate of sulfanilic acid (SA), a model of polar drugs, was enhanced. The relationship between Pcl/PNa and SA flux rate after pretreatment with EDTA was demonstrated to yield a curve asymptotic to the free mobility ratio of those ions. When bile salts were used as the pretreatment agent, instead of EDTA, similar results to the case of EDTA were obtained. Assuming that EDTA interacts with the tight junctional portion of the mucosal membrane to enhance the paracellular permeability to drugs selectively, as demonstrated in many other reports, changes in PCl/PNa and Rm were assumed to correspond to structural change in the tight junction. Consequently, we suggest that these factors can be useful parameters when considering the mechanism of enhanced permeability of the intestinal membrane by several adjuvants.

Adjuvants, Pharmaceutic↗