Urothelial cell transplantation using biodegradable synthetic scaffolds

Reconstruction of the urinary bladder with bowel to restore storage capacity is associated with significant complications arising from substituting an absorptive, mucus-producing intestinal epithelium for the barrier urothelium of the bladder. To overcome these problems, we are developing a "composite enterocystoplasty" procedure to replace the epithelium of the bowel with autologous in vitro-propagated normal urothelial cells. The aims of this study were to evaluate synthetic biomaterials as delivery vehicles for the cultured urothelial cells and provide support during transfer and cell adherence to the de-epithelialized bowel wall. The surgical compliance of 12 biomaterials was evaluated, along with their ability to support urothelial cell attachment. Transfer of urothelial cells onto biomaterials as single cell suspensions or intact cell sheets was investigated. Seeding of a single cell suspension on to non-woven mesh resulted in poor cell attachment. Seeding onto woven mesh was more efficient, but the most effective transfer method involved producing an intact cell sheet that could be combined with woven, knitted and non-woven biomaterials. Transfer of the cell sheet : mesh complexes onto a de-epithelialized bladder stroma produced a stratified epithelium incorporating the strands of the mesh and expressing urothelial-associated antigens after 48 h in organ culture. Thus, we have developed and evaluated a suitable transfer method for in vitro propagated urothelial cells to be used in "composite enterocystoplasty".     

Effect of chitosan glutamate,carbomer 974P,and EDTA on the in vitro Caco-2 permeability and oral pharmacokinetic profile of acyclovir in rats

Background: Chitosan glutamate and polyacrylic acid (e.g., carbomer 974P) are known to modulate the tight junctions in the intestinal wall and increase permeability and blood exposure of drugs absorbed orally by the paracellular route. Aim: To assess the impact of chitosan glutamate and carbomer 974P on the absorption of paracellularly absorbed model drug, acyclovir, in vitro and in rat in vivo. Methods: The influence of chitosan glutamate and carbomer 974P (alone and in combination with EDTA–Na2) on the in vitro Caco-2 permeability and oral pharmacokinetic profile in the rat of acyclovir was investigated. Results: In the presence of chitosan glutamate, the apparent permeability of acyclovir across Caco2 monolayer increased 4.1 times relative to control. This increase was accompanied by a significant (～60%) decrease in transepithelial electrical resistance values indicating opening of the tight junctions in the cell monolayer. In rat, chitosan glutamate doubled oral bioavailability of acyclovir and tripled the amount of acyclovir excreted unchanged into urine. In contrast, the effect of carbomer 974P was not statistically significant at 5% level. Conclusions: In conclusion, chitosan glutamate (1–3%) and chitosan glutamate (1%)/EDTA–Na2 (0.01%) are effective excipients to increase permeability of acyclovir across Caco-2 cell monolayers and the oral absorption in the rat in vivo.     

Formulation of sage essential oil (Salvia officinalis,L.) monoterpenes into chitosan hydrogels and permeation study with GC-MS analysis

This study deals with the formulation of natural drugs into hydrogels. For the first time, compounds from the sage essential oil were formulated into chitosan hydrogels. A sample preparation procedure for hydrophobic volatile analytes present in a hydrophilic water matrix along with an analytical method based on the gas chromatography coupled with the mass spectrometry (GC-MS) was developed and applied for the evaluation of the identity and quantity of essential oil components in the hydrogels and saline samples. The experimental results revealed that the chitosan hydrogels are suitable for the formulation of sage essential oil. The monoterpene release can be effectively controlled by both chitosan and caffeine concentration in the hydrogels. Permeation experiment, based on a hydrogel with the optimized composition [3.5% (w/w) sage essential oil, 2.0% (w/w) caffeine, 2.5% (w/w) chitosan and 0.1% (w/w) Tween-80] in donor compartment, saline solution in acceptor compartment, and semi-permeable cellophane membrane, demonstrated the useful permeation selectivity. Here, (according to lipophilicity) an enhanced permeation of the bicyclic monoterpenes with antiflogistic and antiseptic properties (eucalyptol, camphor and borneol) and, at the same time, suppressed permeation of toxic thujone (not exceeding its permitted applicable concentration) was observed. These properties highlight the pharmaceutical importance of the developed chitosan hydrogel formulating sage essential oil in the dermal applications.     

Response surface method applied to optimization of estradiol permeation in chitosan membranes

The present work deals with the study of estradiol permeation in chitosan membranes. A fractional factorial design was built for the determination of the main factors affecting estradiol permeation. The independent factors analysed were: concentration of chitosan, concentration of cross-linking agent, cross-linking time and thermal treatment. It was found that concentration of chitosan and cross-linking time significantly affected the response. The effects of thermal treatment and concentration of cross-linking agent were not significant. An optimization process based on response surface methodology was carried out in order to develop a statistical model which describes the relationship between active independent variables and estradiol flux. This model can be used to find out a combination of factor levels during response optimization. Possible options for response optimization are to maximize, minimize or move towards a target value.     

Transdermal delivery of ketorolac tromethamine: effects of vehicles and penetration enhancers

The effects of vehicles and penetration enhancers on the in vitro permeation of ketorolac tromethamine (KT) across excised hairless mouse skins were investigated. Among pure vehicles examined, propylene glycol monolaurate (PGML) showed the highest permeation flux, which was 94.3 ± 17.3 µg/cm²/h. Even though propylene glycol monocaprylate (PGMC) alone did not show high permeation rate, the skin permeability of KT was markedly increased by the addition of diethylene glycol monoethyl ether (DGME); the enhancement factors were 19.0 and 17.1 at 20% and 40% of DGME, respectively. When DGME was added to PGML, the permeation fluxes were almost two times at 20-60% of DGME compared to PGML alone. In the cosolvent system consisting of propylene glycol (PG)-oleyl alcohol, the permeation rate increased as the ratio of PG increased. In the study to investigate the effect of drug concentration on the permeation rate of KT, the permeation rates increased as the drug concentration increased in all vehicles used, and the dramatic increase in permeation rate was obtained when the drug concentration was higher than its solubility. For the effects of fatty acids on the permeation of KT, five fatty acids were added to PG at concentrations of 1%-, 3%-, 5%- and 10%-caprylic acid, capric acid, lauric acid, oleic acid, and linoleic acid. The enhancing effects of fatty acids were different, depending on the concentration as well as the sort of fatty acids. The highest enhancing effect was attained with 10% caprylic acid in PG; the permeation flux was 113.6 ± 17.5 µg/cm²/h. The lag time of KT was reduced as the concentration of fatty acids increased except for caprylic acid.     

The impact of preparation parameters on typical attributes of chitosan-heparin nanohydrogels: particle size,loading efficiency,and drug release

Today, developing an optimized nanoparticle (NP) preparation procedure is of paramount importance in all nanoparticulate drug delivery researches, leading to expanding more operative and clinically validated nanomedicines. In this study, a one-at-a-time experimental approach was used for evaluating the effect of various preparation factors on size, loading, and drug release of hydrogel NPs prepared with ionotropic gelation between heparin and chitosan. The size, loading efficiency (LE) and drug release profile of the NPs were evaluated when the chitosan molecular weight, chitosan concentration, heparin addition time to chitosan solution, heparin concentration, pH value of chitosan solution, temperature, and mixing rate were changed separately while other factors were in optimum condition. The results displayed that size and LE are highly influenced by chitosan concentration, getting an optimum of 63?±?0.57 and 75.19?±?2.65, respectively, when chitosan concentration was 0.75?mg/ml. Besides, heparin addition time of 3?min leaded to 74.1?±?0.79 % LE with no sensible effect on size and release profile. In addition, pH 5.5 showed a minimum size of 63?±?1.87, maximum LE of 73.81?±?3.13 and the slowest drug release with 63.71?±?3.84 % during one week. Although LE was not affected by temperature, size and release reduced to 63?±?0 and 74.21?±?1.99% when temperature increased from 25°C to 55°C. Also, continuous increase of mixer rate from 500 to 3500?rpm resulted in constant enhancement of LE from 58.3?±?3.6 to 74.4?±?2.59 as well as remarkable decrease in size from 148?±?4.88 to 63?±?2.64.     

Gramicidin channels

Gramicidin channels are mini-proteins composed of two tryptophan-rich subunits. The conducting channels are formed by the transbilayer dimerization of nonconducting subunits, which are tied to the bilayer/solution interface through hydrogen bonds between the indole NH groups and the phospholipid backbone and water. The channel structure is known at atomic resolution and the channel's permeability characteristics are particularly well defined: gramicidin channels are selective for monovalent cations, with no measurable permeability to anions or polyvalent cations; ions and water move through a pore whose wall is formed by the peptide backbone; and the single-channel conductance and cation selectivity vary when the amino acid sequence is varied, even though the permeating ions make no contact with the amino acid side chains. Given the amount of experimental information that is available-for both the wild-type channels and for channels formed by amino acid-substituted gramicidin analogues-gramicidin channels provide important insights into the microphysics of ion permeation through bilayer-spanning channels. For the same reason, gramicidin channels constitute the system of choice for evaluating computational strategies for obtaining mechanistic insights into ion permeation through the complex channels formed by integral membrane proteins.     

Permeation and skin retention of quercetin from microemulsions containing Transcutol® P

A microemulsion for the cutaneous release of quercetin was prepared. An aqueous phase, containing 40% Transcutol(?) P as solubilizing agent and permeation enhancer, was emulsified with Labrafil(?) as oil phase and Labrasol(?)/Capryol(?) 90 as Solvent/Co-solvent. Quercetin was dissolved in the microemulsion at the concentration of 1%. Ternary phase diagrams were generated to determine the optimal concentration of each excipient composing the microemulsion. The physicochemical properties of the microemulsion, such as pH, viscosity, refractive index, and particle size distribution were determined. The microemulsion was stable for 12 months at the storing conditions of 25.0 ± 1.0°C. The in vitro quercetin permeability into and through the abdominal hairless pig skin was determined by vertical Franz's cells. Quercetin showed hardly any permeability through the skin when dissolved in water- and Transcutol(?) P-free media, whereas a remarkable increase in cutaneous permeability was observed when quercetin was formulated in the microemulsion or when simply dissolved in Transcutol(?) P. These two last formulations are those showing the lower skin retention.     

In situ oil/water separation using hydrophobic-oleophilic fibrous wall: a lab-scale feasibility study for groundwater cleanup

Kapok, a natural plant fiber, possesses excellent hydrophobic-oleophilic characteristics. Its innovative use as hydrophobic-oleophilic wall that allows permeation of oil but not water into an oil recovery well is proposed. Its performance was investigated through laboratory experiments, in which diesel was used as the experimental oil. A two-dimensional hydraulic flume was setup to physically model the oil/water separation by the kapok wall. The influences of packing density, kapok wall thickness and oil thickness on the oil recovery rate were examined. The oil permeability of the packed kapok decreased from 0.0165 cm2 at 34 g/L packing density to 0.0038 cm2 at 70 g/L packing density. The kapok wall exhibited complete rejection of water while allowed oil to permeate through. The excellent oil/water separation by the kapok wall was due to surface interaction between the kapok fibers and the oil, which resulted in spontaneous penetration and permeation of the oil through the kapok wall. The oil recovery rate increased with thickness of the oil layer in the feed stream. When the oil thickness exceeded 60 mm, a constant flux of 3.8-5.0, 3.2-3.3 and 2.5-2.7 L/(m2 min) could be achieved by the kapok wall of 55-, 75- and 95-mm thick, respectively, under the natural pressure gradient. The kapok wall could be reused for several wetting/drying cycles, and only lost 27% of its initial oil permeability.     

醋酸对明胶-壳聚糖膜性质的影响及应用

目的制备基于不同醋酸质量分数的明胶-壳聚糖基可食复合膜,以期探究醋酸质量分数(1%、1.5%、2%和2.5%)对复合膜性质及应用的影响。方法利用透湿仪、透氧仪、拉伸仪、扫描电镜、热重、红外和拉曼光谱探究可食性复合膜的理化性质,并进行机理解析。结果低醋酸浓度制备的复合膜具有较低透湿性和透氧性,且其抗拉强度和断裂伸长率较高;同时醋酸浓度较低时,可以提高复合膜表面的光滑度,改善复合膜的热稳定性。红外测试与拉曼测试表明,明胶、壳聚糖、醋酸之间主要是通过氢键进行相互作用并结合,从而形成了稳定致密的薄膜。结论以质量分数为1%醋酸制备得到的明胶-壳聚糖基复合膜,其性质表现得更加优异,同时该系列薄膜可以制备成包装袋并能有效包装花生,对明胶-壳聚糖复合膜在食品包装领域的应用具有一定的参考作用。     

Development of a sustained release dosage form for alpha-lipoic acid. I. Design and in vitro evaluation

The purpose of this study was the design and evaluation of a sustained release dosage form for the oral administration of alpha-lipoic acid. The cationic polymer chitosan was used in order to provide a controlled drug release based on ionic interactions with the anionic drug. The effect of such ionic interactions on the release of alpha-lipoic acid could be verified by diffusion studies. In vitro release studies with tablets (diameter: 10.0 mm; thickness: approximately 4 mm) containing 80% alpha-lipoic acid and 20% chitosan acetate showed a controlled drug release over a time period of 24 h. Raising the ratio of chitosan acetate in such delivery systems led to an even stronger retardation of drug release. In addition, permeation studies carried out in Ussing-type chambers with freshly excised intestinal mucosa from guinea pigs demonstrated no significant (p < 0.05) influence of the degree of drug ionization on its absorption behavior. The apparent permeability coefficient (Papp) for alpha-lipoic acid was determined to be 1.39 +/- 0.28 x 10(-5) cm/sec at pH 6.4 (means +/- SD). The use of a sustained delivery system for alpha-lipoic acid, which is based on ionic interactions, should therefore have no influence on the absorption behavior of the drug. The sustained release dosage forms described here might provide a constant plasma level of the drug being highly beneficial for various therapeutic reasons.     

Nano-engineered microcapsules of tannic acid and chitosan for protein encapsulation

Tannic acid (TA), a high molecular weight polyphenol of natural origin, was assembled in alternation with chitosan (CH) using a layer-by-layer technique. The deposition of tannic acid and chitosan layers on flat supports was monitored by quartz crystal microbalance, UV-vis spectroscopy, and electrophoretic mobility measurements on microparticles. Hollow (TA/CH)4 capsules were built and their permeability as a function of pH and molecular weight of a penetrating compound was investigated. The pH-permeability threshold for TA/CH capsules is shifted to lower pH for 2 pH units, as compared with commonly used polyallylamine/polystyrene sulfonate capsules. A more pronounced dependence of the TA/CH capsules' permeability on molecular weight of encapsulated substances allows better control over their release properties. Bovine serum albumin was loaded into (TA/CH)4 capsules using a pH-driven method and released by decreasing pH. Biocompatible tannic acid/chitosan films and capsules have advantages toward capsules made of synthetic polyelectrolytes for drug encapsulation and as delivery and depot systems. Incorporating a layer of tannic acid with proved antioxidant and antimicrobial properties into capsule walls, provides defense for encapsulated materials.     

Long-Term Skin Permeation Kinetics of Estradiol (I): Effect of Drug Solubilizer-Polyethylene Glycol 400

Abstract

A skin permeation cell was recently developed to overcome the deficiencies noted in the currently available in vitro diffusion cells, and to provide a cell design which is suitable for studying the long-term drug permeation kinetics through the skin and is also sensitive enough for assessing the mechanisms of skin permeation by a high performance liquid chromatography.

To evaluate the rote of drug reservoir concentration in the kinetics of skin permeation as well as to maintain a sink condition in the receptor solution, the water-miscible polyethylene glycol (PEG) 400 was incorporated into the saline solution to act as a solubilizer to enhance the aqueous solubility of the relative water-insoluble estradiol. The equilibrium solubility of estradiol at 37°C was observed to in crease exponentially as increasing the volume fraction of PEG 400 added.

The rates o f permeation of estradiol across the male and female hairless mouse, whole and stripped skins excised freshly from the abdominal region, were measured a t various PEG concentrations and the permeability coefficients were determined. The permeability co-efficients were found t o decrease as increasing the PEG concentration. A linear relationship was established between th e permeability co-efficients and the skin /solution partition coefficients and the steady-stated if fusivity was calculated. Effect of sex was assessed.

The rate of permeation and the permeability coefficient across the stratum corneum were determined, using t h e multi-laminated dif-fusional resistance model. Results demonstrated that the stratum corneum acts as the rate-limiting barrier in the skin permeation of estradiol and the incorporation of upto 40% v/v PEG 400 does not in-fluence the barrier propertiesof stratum corneum, even though PEG 400 has been found to affect the aqueous solubility, permeability co-efficient, and skin /solution partition coefficient of estradiol.     

Long-Term Skin Permeation Kinetics of Estradiol (I): Effect of Drug Solubilizer-Polyethylene Glycol 400

A skin permeation cell was recently developed to overcome the deficiencies noted in the currently available in vitro diffusion cells, and to provide a cell design which is suitable for studying the long-term drug permeation kinetics through the skin and is also sensitive enough for assessing the mechanisms of skin permeation by a high performance liquid chromatography.

To evaluate the rote of drug reservoir concentration in the kinetics of skin permeation as well as to maintain a sink condition in the receptor solution, the water-miscible polyethylene glycol (PEG) 400 was incorporated into the saline solution to act as a solubilizer to enhance the aqueous solubility of the relative water-insoluble estradiol. The equilibrium solubility of estradiol at 37°C was observed to in crease exponentially as increasing the volume fraction of PEG 400 added.

The rates o f permeation of estradiol across the male and female hairless mouse, whole and stripped skins excised freshly from the abdominal region, were measured a t various PEG concentrations and the permeability coefficients were determined. The permeability co-efficients were found t o decrease as increasing the PEG concentration. A linear relationship was established between th e permeability co-efficients and the skin /solution partition coefficients and the steady-stated if fusivity was calculated. Effect of sex was assessed.

The rate of permeation and the permeability coefficient across the stratum corneum were determined, using t h e multi-laminated dif-fusional resistance model. Results demonstrated that the stratum corneum acts as the rate-limiting barrier in the skin permeation of estradiol and the incorporation of upto 40% v/v PEG 400 does not in-fluence the barrier propertiesof stratum corneum, even though PEG 400 has been found to affect the aqueous solubility, permeability co-efficient, and skin /solution partition coefficient of estradiol.     

有机酸溶剂体系中壳聚糖膜性能的研究进展

目的 有机酸溶剂体系对壳聚糖的成膜性能至关重要,探讨其潜在的分子机制,可为壳聚糖膜的相关研究提供理论参考。方法 综述壳聚糖的化学组成和成膜机制,以及壳聚糖在不同有机酸溶剂体系中成膜的力学性能、溶胀性、氧气透过率、水蒸气透过率等特性。结果 有机酸溶剂体系可为壳聚糖的溶解提供更多的质子,从而促进壳聚糖分子在水中的溶解。此外,有机酸分子结构中羧酸和羟基数量、碳链长度不同,其与壳聚糖分子间形成的氢键和离子作用力不同,进而可在很大程度上影响壳聚糖的成膜性能。结论 有机酸溶剂体系对壳聚糖膜的性能具有较为显著的影响,这有助于提高其作为可食用膜在改善食品品质和保鲜方面的适用性。     

Synthesis and Skin Permeation Study of Lidocaine Organic Salts

Four organic quarternary ammonium salts of lidocaine, lidocaine adipate, lidocaine maleate, lidocaine malonate and lidocaine tosylate were prepared in this study. These compounds were regarded as prodrugs of lidocaine and were expected to enhance skin permeability by ion-pair approach. In vitro permeation study through pig skin and determination of partition coefficients were carried out. The skin permeation of lidocaine adipate and lidocaine malonate after 12 hours of application was significantly higher than lidocaine hydrochloride but was not significantly different from lidocaine (α < 0.05). The increase in permeation of organic salts from lidocaine hydrochloride and the different profile of permeation from lidocaine may be the result of prodrug structure contributing ion-pair transportation beside normal route of transportation.     

In Vitro Assessment of Acyclovir Permeation Across Cell Monolayers in the Presence of Absorption Enhancers

The aim of the investigation was to establish transepithelial permeation of acyclovir across Caco-2 and Madin-Darby canine kidney (MDCK) cell monolayers and attempt to improve its permeation by employing absorption enhancers (dimethyl β cyclodextrin, chitosan hydrochloride and sodium lauryl sulfate) and combinations thereof. Caco-2 and MDCK cell monolayers have been widely employed in studying drug transport, mechanisms of drug transport, and screening of absorption enhancers and excipients. Transepithelial electrical resistance and permeation of ^99mTc-mannitol were employed as control parameters to assess the tight junction and paracellular integrity. Permeation of acyclovir in the presence of absorption enhancers was found to be significantly higher compared with drug permeation in their absence when assessed as apparent permeability coefficients (P_app). Synergistic improvements in P_app values of acyclovir were obtained in case-selected combinations of absorption enhancers; dimethyl β cyclodextrin–chitosan hydrochloride, chitosan hydrochloride–sodium lauryl sulfate, and dimethyl β cyclodextrin–sodium lauryl sulfate, were used. Recovery and viability assessment studies of both cell monolayers suggested reestablishment of paracellular integrity and no damage to cell membranes. Significantly improved permeation of acyclovir in the presence of selected combinations of absorption enhancers may be used as a viable approach in overcoming the problem of limited oral bioavailability of acyclovir.     

Effect of Penetration Enhancers on Transdermal Delivery of Timolol Maleate

In vitro skin permeation of Timolol maleate through human cadaver skin was studied using Franz diffusion cell. The results indicate that the drug penetrates poorly through human cadaver skin. However, skin penetration enhancers such as dimethyl sulfoxide (DMSO), oleic acid (OA) and lauryl chloride (LC) enhanced the permeability of Timolol maleate (TM) through human cadaver skin. The permeation enhancement of drug was maximum by lauryl chloride amongst the three enhancers. Moreover, lauryl chloride increases the permeation of drug through skin with increase in the time of application and concentration on skin. The change in lag time was also observed.     

Preparation and embedding–releasing behavior of polyelectrolyte microcapsule

以高压静电法制备的壳聚糖微球为模板, 与海藻酸钠层层自组装制备多层聚电解质膜, 再以乙酸溶出壳聚糖微球制备出微胶囊. 调节乙酸的浓度及溶出时间以控制核内壳聚糖的残留量, 从而诱导负电性荧光素钠的沉积. 微胶囊内部荧光素钠的浓度随着包埋时间的增加而呈线性地提高, 当荧光素钠溶液的浓度为2 mg/l时其最终包埋的浓度达到10.2 mg/l. 这种聚电解质微囊对于小分子负电荷荧光素钠的释放行为主要受其囊壁和内部模板结构的影响, 盐离子浓度对荧光素钠释放行为的影响较小.     

In vitro assessment of acyclovir permeation across cell monolayers in the presence of absorption enhancers

The aim of the investigation was to establish transepithelial permeation of acyclovir across Caco-2 and Madin-Darby canine kidney (MDCK) cell monolayers and attempt to improve its permeation by employing absorption enhancers (dimethyl β cyclodextrin, chitosan hydrochloride and sodium lauryl sulfate) and combinations thereof. Caco-2 and MDCK cell monolayers have been widely employed in studying drug transport, mechanisms of drug transport, and screening of absorption enhancers and excipients. Transepithelial electrical resistance and permeation of ^99mTc-mannitol were employed as control parameters to assess the tight junction and paracellular integrity. Permeation of acyclovir in the presence of absorption enhancers was found to be significantly higher compared with drug permeation in their absence when assessed as apparent permeability coefficients (P_app). Synergistic improvements in P_app values of acyclovir were obtained in case-selected combinations of absorption enhancers; dimethyl β cyclodextrin-chitosan hydrochloride, chitosan hydrochloride-sodium lauryl sulfate, and dimethyl β cyclodextrin-sodium lauryl sulfate, were used. Recovery and viability assessment studies of both cell monolayers suggested reestablishment of paracellular integrity and no damage to cell membranes. Significantly improved permeation of acyclovir in the presence of selected combinations of absorption enhancers may be used as a viable approach in overcoming the problem of limited oral bioavailability of acyclovir.