Polyelectrolyte Microcapsules: An Extended Release System for the Antiarrhythmic Complex of Allapinin with Glycyrrhizic Acid Salt.

Allapinin has antiarrhythmic activity and can be used to prevent and treat various supraventricular and ventricular arrhythmias. Nevertheless, it is highly toxic and has a number of side effects associated with non-specific accumulation in various tissues. The complex of this substance with the monoammonium salt of glycyrrhizic acid (Al:MASGA) has less toxicity and improved antiarrhythmic activity. However, the encapsulation of Al:MASGA in polyelectrolyte microcapsules (PMC) for prolonged release will reduce the residual adverse effects of this drug. In this work, the possibility of encapsulating the allapinin-MASGA complex in polyelectrolyte microcapsules based on polyallylamine and polystyrene sulfonate was investigated. The encapsulation methods of the allapinin-MASGA in polyelectrolyte microcapsules by adsorption and coprecipitation were compared. It was found that the coprecipitation method did not result in the encapsulation of Al:MASGA. The sorption method facilitated the encapsulation of up to 80% of the original substance content in solution in PMC. The release of the encapsulated substance was further investigated, and it was shown that the release of the encapsulated Al:MASGA was independent of the substance content in the capsules, but at pH 5, a two-fold decrease in the rate of drug release was observed.

Determination of Phenol with Peroxidase Immobilized on CaCO3.

Phenols are widely used in industries despite their toxicity, which requires governments to limit their concentration in water to 5 mg/L before discharge to the city sewer. Thus, it is essential to develop a rapid, simple, and low-cost detection method for phenol. This study explored two pathways of peroxidase immobilization to develop a phenol detection system: peroxidase encapsulation into polyelectrolyte microcapsules and peroxidase captured by CaCO3. The encapsulation of peroxidase decreased enzyme activity by 96%; thus, this method cannot be used for detection systems. The capturing process of peroxidase by CaCO3 microspherulites did not affect the maximum reaction rate and the Michaelis constant of peroxidase. The native peroxidase-Vmax = 109 µM/min, Km = 994 µM; CaCO3-peroxidase-Vmax = 93.5 µM/min, Km = 956 µM. Ultimately, a reusable phenol detection system based on CaCO3 microparticles with immobilized peroxidase was developed, capable of detecting phenol in the range of 700 ng/mL to 14 µg/mL, with an error not exceeding 5%, and having a relatively low cost and production time. The efficiency of the system was confirmed by determining the content of phenol in a paintwork product.

Sorption of Salts of Various Metals by Polyelectrolyte Microcapsules.

Anthropogenic activity negatively affects the environment by polluting it with the salts of various metals. One of the ways to reduce this influence is to use water purification methods for the salts of various metals. Water purification methods based on nanomaterials are promising. In this regard, we proposed to study polyelectrolyte microcapsules (PMC) as a promising sorption agent for the salts of various metals. It was found that the polystyrene sulfonate-polyallylamine (PSS-PAH) polyelectrolyte complex and polyelectrolyte microcapsules of different compositions are not able to adsorb salts CuSO4, Pb(NO)3, FeCl3, and CuCl2. At the same time, it was found that all types of capsules, except for (PSS/PAH)2/PSS, are capable of sorbing about 420 µg of K3[Fe(CN)6] and about 500 µg of K4[Fe(CN)6] from solution. The adsorption of polyelectrolyte microcapsules has an electrostatic nature which is confirmed by increases in the sorption capacity of PMC of K3[Fe(CN)6] and K4[Fe(CN)6] with decreases in the pH of the solution. Also, It was confirmed that the sorption process of PMC of K3[Fe(CN)6] and K4[Fe(CN)6] is concentration dependent and has the limitation of the number of binding sites.

Polyelectrolyte Microcapsules-A Promising Target Delivery System of Amiodarone with the Possibility of Prolonged Release.

Atrial fibrillation is one of the most common cardiac arrhythmias. Pharmacological preparations are used for treatment to control heart rate and rhythm. Amiodarone is one of these highly effective preparations, but, at the same time, it has significant toxicity and nonspecific accumulation in tissues. The drug delivery system based on polyelectrolyte microcapsules is one of the solutions. For this purpose, we compared different encapsulation methods of amiodaron: monoammonium salt of glycyrrhizic acid (Am:MASGA) complex (molar ratio 1:8). The concentration of amiodarone was determined by spectrophotometric methods at 251 nm. It has been shown that the co-precipitation method allows capturing 8% of Am:MASGA by CaCO3 microspherulites, which is not sufficient for the long-acting drug. The adsorption method allows encapsulating more than 30% of Am:MASGA into CaCO3 microspherulites and polyelectrolyte microcapsules CaCO3(PAH/PSS)3, but, at the same time, an insignificant amount of substance is released into the incubation medium. The development of delivery and long-acting drug system based on such methods are not inexpedient. The most appropriate encapsulation method of Am:MASGA is the adsorption method into polyelectrolyte microcapsules with complex interpolyelectrolyte structure (PAH/PSS)3. Such a type of PMC adsorbed about 50% of the initial amount of the substance and 25-30% of Am:MASGA was released into the medium after 115 h of incubation. The adsorption of Am:MASGA by polyelectrolyte microcapsules has electrostatic nature as evidenced by the acceleration of the release by 1.8 times as ionic strength increases.

A Study of the Buffer Capacity of Polyelectrolyte Microcapsules Depending on Their Concentration and the Number of Layers of the Polyelectrolyte Shell.

Polyelectrolyte microcapsules are used in the development of new forms of targeted delivery systems, self-healing materials, sensors, and smart materials. Nevertheless, their buffer capacity has not been practically studied, although that characteristic makes it possible to estimate the change in the state of protonation of the entire polyelectrolyte system. This is necessary both for creating a buffer barrier system for pH-sensitive compounds (metals, enzymes, polyelectrolytes, drugs) and for the correct interpretation of the results of research and studying of the PMC structure. The buffer capacity of a PMC can be affected by the concentration of microcapsules in solution and the number of shell layers since the listed parameters affect other physicochemical properties of the PMC shell. This includes, for example, the electrical conductivity, permeability (of ions), osmotic pressure, charge density, etc. In this regard, we studied the change in the buffer capacity of polyelectrolyte microcapsules depending on their concentration and the number of shell layers. As a result, it was found that with an increasing concentration of microcapsules, the buffering capacity of the PMC increases, but at the same time, in the pH range from 4 to 5.5, the calculated buffering capacity of 1 billion capsules decreases with increasing their concentration. This effect may be associated with a decrease in the available -NH2 groups of the PMC's shell. In addition, it was found that the main contribution to the buffer capacity of a PMC is made by the entire shell of the microcapsule and not just its surface. At the same time, the buffer capacity of the capsules has non-linear growth with an increase in the number of PMC shell layers. It is presumably associated either with a decrease in the polyelectrolyte layer with an increase in their number or with a decrease in the permeability of hydrogen protons.

A Study of the Buffer Capacity of Polyelectrolyte Microcapsules Depending on Their Ionic Environment and Incubation Temperature.

Polyelectrolyte microcapsules (PMCs) are used in the development of new forms of drugs, coatings and diagnostic systems. Their buffer capacity, depending on the conditions of the medium, has not been practically studied, although it can affect the structure of both the capsule itself and the encapsulated agents. In this connection, we studied the buffer capacity of polyelectrolyte microcapsules of the composition (polystyrene sulfonate/polyallylamine)3 ((PSS/PAH)3) depending on the concentration and the type of salt in solution, as well as the microcapsule incubation temperature. It was found that the buffer capacity of microcapsules in the presence of mono- and di-valent salts of the same ionic strength did not differ practically. Increasing the NaCl concentration to 1 M led to an increase of buffer capacity of PMCs at pH ≥ 5, and an increase in NaCl concentration above 1 M did not change buffer capacity. The study of the buffer capacity of pre-heated PMCs showed that buffer capacity decreased with increasing incubation temperature, which was possibly due to the compaction of the PMCs and an increase in the number of compensated PAH sites. The addition of 1 M sodium chloride to heated PMCs presumably reversed the process described above, since an increase in the ionic strength of the solution led to an increase of the buffer capacity of the PMCs. The effects described above confirm the hypothesis put forward that the buffer properties of microcapsules are determined by uncompensated PAH regions in their composition.

Destruction of polyelectrolyte microcapsules and release of FITC-dextran from them by the influence of sodium dodecyl sulfonate.

Polyelectrolyte microcapsules can be applied as microcontainers for the delivery of a wide range of substances, and it is important to search for new methods for capsule destruction and releasing substances from them. In this work, we studied the possibility of using sodium dodecyl sulfonate (SDS) for the release of fluorescein isothiocyanate-dextran from six-layer microcapsules composed of PAH and PSS. It was shown that the presence of SDS in the medium, at a concentration of 3000 µg/ml, leads to the destruction of polyelectrolyte microcapsules and the release of the substance from them (54% of the amount of the encapsulated substance), while the main part of the FITC-dextran released during the first hours of incubation. At an SDS concentration of 100 µg/ml, the substance released is uniform and is 44% in 24 h. At SDS concentrations from 50 to 100 µg/ml, the process of destruction of microcapsules proceeds more slowly. At SDS concentrations from 10 to 50 µg/ml, microcapsules are not degraded.

The Pathways to Create Containers for Bacteriophage Delivery.

Antimicrobial resistance is a global public health threat. One of the possible ways to solve this problem is phage therapy, but the instability of bacteriophages hinders the development of this approach. A bacteriophage delivery system that stabilizes the phage is one of the possible solutions to this problem. This study is dedicated to exploring methods to create encapsulated forms of bacteriophages for delivery. We studied the effect of proteolytic enzymes on the destruction of the polyelectrolyte microcapsule shell and revealed that protease from Streptomyces griseus was able to destroy the membrane of the microcapsule (dextran sulfate/polyarginine)3 ((DS/PArg)3). In addition, the protease decreased the activity of the bacteriophage in the second hour of incubation, and the phage lost activity after 16 h. It was found that a medium with pH 9.02 did not affect the survival of the bacteriophage or E. coli. The bacteriophages were encapsulated into polyelectrolyte microcapsules (DS/PArg)3. It was established that it is impossible to use microcapsules as a means of delivering bacteriophages since the bacteriophages are inactivated. When bacteriophages were included inside a CaCO3 core, it was demonstrated that the phage retained activity before and after the dissolution of the CaCO3 particle. From the results of this study, we recommend using CaCO3 microparticles as a container for bacteriophage delivery through the acidic stomach barrier.

Qualitative and quantitative methods detection of SDS based on polyelectrolyte microcapsules.

Sodium dodecyl sulfate (SDS) is the most widely used anionic surfactant. Its frequent use causes environmental pollution and negative effects on living organisms (even at low concentrations ≈ 20 µg/ml). Thus, cheap and fast methods are needed to detect this surfactant in wastewater and surface waters in order to prevent the negative effects of SDS on the environment and human beings. We discovered that sodium dodecyl sulfate is capable of destroying polyelectrolyte microcapsules, which has been demonstrated by the number of sedimented polyelectrolyte microcapsules (PMC) before and after incubation in SDS solution. Therefore, it was proposed to use PMCs to create qualitative and quantitative diagnostic systems for the determination of SDS in solution. The qualitative system is a polyelectrolyte microcapsules containing polyallylamine labeled with a fluorescent dye-FITC. An excess SDS concentration of more than 5 µg/ml in the analyzed medium leads to the destruction of PMC and an increase in the fluorescence intensity of the solution, which is recorded by a fluorometer. The quantitative diagnostic system is based on turbidimetry of the PMC suspension before and after incubation in an anionic surfactant solution. This system has a range of detectable SDS concentrations from 10 to 50 µg/ml, with a standard deviation of no more than 11%.

The Discovery of the Buffer Capacity of Various Types of Polyelectrolyte Microcapsules.

Polyelectrolyte microcapsules, which are obtained by the method of alternate adsorption of oppositely charged polyelectrolytes onto colloidal particles of micron size, are widely used in science and industry. Nevertheless, the properties of microcapsules are still poorly understood. In particular, there is no information in the literature on the buffer capacity. However, information on the presence of a buffer capacity and an understanding of its mechanisms can both simplify the use of microcapsules and expand the scope of their application. In this regard, the buffer capacity of various types of microcapsules was studied. It was found that polyelectrolyte microcapsules consisting of polyallylamine, and polystyrene sulfonate have a buffer capacity. In addition, in an acidic medium, the buffer capacity of microcapsules containing BSA is significantly greater than that of microcapsules without protein. This is due to the fact that BSA contributes to the buffering of microcapsules. Differences in the behaviour of the buffer capacity of microcapsules with the composition (PAH/PSS)3 and (PSS/PAH)3 were found. In addition, a hypothesis has been proposed that regions of unbound polyallylamine are responsible for the buffering properties of polyelectrolyte microcapsules. This hypothesis is confirmed by the fact that incubation of microcapsules in 0.5 M NaCl increases the amount of unbound polyallylamine, which leads to an increase in the buffer capacity of microcapsules at alkaline pH values higher than the buffer capacity of capsules in an aqueous solution.

New sight at the organization of layers of multilayer polyelectrolyte microcapsules.

In this work, the mutual arrangement of polyelectrolytes of multilayer polyelectrolyte microcapsules (with layers-[PAH/PSS]3PAH) by determination of the dissociation level of polyallylamine (PAH) from the surface of a polyelectrolyte microcapsules (PMC) of various types was studied: PMC with a dissolved CaCO3 core after preparation, PMC with an undissolved CaCO3 core and PMC with an encapsulated protein. It was concluded that the polyelectrolyte layers are mixed in the entire shell of the capsules with a dissolved CaCO3 core. In the case of the PMC with an undissolved CaCO3 core, such mixing of polyelectrolyte layers does not occur. That fact allows us to conclude that the mixing of polyelectrolytes layers mixing at the stage of dissolution of CaCO3 core. The PMC with encapsulated protein has partial mixing of polyelectrolytes layers. That phenomenon may be due to the fact that seven-layered protein-containing microcapsules already have a dense and well-formed shell. The obtained data correlate with the data on the study of the surface charge of microcapsules.

The Influence of Polyanions and Polycations on Bacteriophage Activity.

Phage therapy is a great alternative to antibiotic drugs, but it can't effectively overcome the over-acidic medium of the stomach. We offer the use of polyelectrolyte microcapsules as a protective means of bacteriophage. It is necessary to understand the influence of polyelectrolytes on bacteriophage survival. The work studied the effect of polyanions and polycations on the coliprotetic bacteriophage's viability. We have shown that polyallylamine decreased bacteriophage's viability during increasing polyelectrolyte concentration and polyarginine had a lower inhibitory effect (then PAH) on the activity of the bacteriophage due to polyelectrolyte concentration from 0.05 to 5 mg/mL. It was shown that the inhibition of the bacteriophage by polyallylamine had an electrostatic nature and the use of high ionic strength prevented the formation of the PAH-protein capsid complex. Polystyrene sulfonate does not affect bacteriophage viability during increasing polyelectrolyte concentration from 0.05 mg/mL to 1 mg/mL. Polystyrene sulfonate decreases the viability of bacteriophage from 5 mg/mL of polyelectrolyte concentration. Dextran sulfate inhibits bacteriophage activity at 20-30%. Dextran inhibits bacteriophage activity by 80% at diapason concentration from 0.05 to 5 mg/mL and loses the inhibition effect from a concentration of 5 mg/mL.

Effect of Pollyallylamine on Alcoholdehydrogenase Structure and Activity.

In this article, the effect of polyallylamine (PAA) on the structure and catalytic characteristics of alcohol dehydrogenase (ADH) was studied. For this research, we used methods of stationary kinetics and fluorescence spectroscopy. It has been shown that PAA non-competitively inhibits ADH activity while preserving its quaternary structure. It was established that 0.1 M ammonium sulfate removes the inhibitory effect of PAA on ADH, which is explained by the binding of sulfate anion (NH4)2SO4 with polyallylamine amino groups. As a result, the rigidity of the polymer chain increases and the ability to bind to the active loop of the enzyme increases. It is also shown that sodium chloride removes the inhibitory effect of PAA on ADH due to an electrostatic screening of the enzyme from polyelectrolyte. The method of encapsulating ADH in polyelectrolyte microcapsules was adapted to the structure and properties of the enzyme molecule. It was found that the best for ADH is its encapsulation by adsorption into microcapsules already formed on CaCO3 particles. It was shown that the affinity constant of encapsulated alcohol dehydrogenase to the substrate is 1.7 times lower than that of the native enzyme. When studying the affinity constant of ADH in a complex with PAA to ethanol, the effect of noncompetitive inhibition of the enzyme by polyelectrolyte was observed.

Destruction of Polyelectrolyte Microcapsules Formed on CaCO3 Microparticles and the Release of a Protein Included by the Adsorption Method.

The degradation of polyelectrolyte microcapsules formed on protein-free CaCO3 particles consisting of polyallylamine (PAH) and polystyrene sulfonate (PSS) and the resulting yield of protein in the presence of various salts of different concentrations, as well as at two pH values, was studied by fluorescence spectroscopy; the protein was incorporated into prepared microcapsules by adsorption. It was found that a high concentration of sodium chloride (2 M) leads to considerable dissociation of PAH, which is apparently due to the loosening of polyelectrolytes under the action of ionic strength. At the same time, 0.2 M sodium chloride and ammonium sulfate of the same ionic strength (0.1 M) exert less influence on the amount of dissociated polymer. In the case of ammonium sulfate (0.1 M), the effect is due to the competitive binding of sulfate anions to the amino groups of the polyelectrolyte. However, unlike microcapsules formed on CaCO3 particles containing protein, the dissociation of polyelectrolyte from microcapsules formed on protein-free particles increased with increasing temperature. Apparently, a similar effect is associated with the absence of a distinct shell, which was observed on microcapsules formed on protein-containing CaCO3 particles. The high level of the presence of Fluorescein isothiocyanate (FITC)-labeled Bovine Serum Albumin (BSA) in the supernatant is explained by the large amount of electrostatically bound protein and the absence of a shell that prevents the release of the protein from the microcapsules. In 2M NaCl, during the observation period, the amount of the released protein did not exceed 70% of the total protein content in the capsules, in control samples, this value does not exceed 8%, which indicates the predominantly electrostatic nature of protein retention in capsules formed on protein-free CaCO3 particles. The increase in protein yield and peeling of PAH with increasing pH is explained by the proximity of pH 7 to the point of charge exchange of the amino group of polyelectrolyte, as a result of the dissociation of the microcapsule.

Decapsulation of Dextran by Destruction of Polyelectrolyte Microcapsule Nanoscale Shell by Bacillus subtilis Bacteria.

One of the prerequisites of successful address delivery is controlling the release of encapsulated drugs. The new method of bacterial spore encapsulation in polyelectrolyte microcapsules allows for degrading the nanoscale membrane shell of microcapsules. The possibility of encapsulating spore forms of Bacillus subtilis in polystyrenesulfonate sodium/ polyallylamine hydrochloride (PSS/PAH) polyelectrolyte microcapsules was demonstrated. The activation and growth on a nutrient medium of encapsulated bacterial spores led to 60% degradation of the microcapsules nanoscale membrane shell. As a result, 18.5% of Fluorescein isothiocyanatedextran was encapsulated into polyelectrolyte microcapsules, and 28.6% of the encapsulated concentration of FITC-dextran was released into the solution.

Comparative Study of Electrochemical Sensors Based on Enzyme Immobilized into Polyelectrolyte Microcapsules and into Chitosan Gel.

The characteristics of an electrochemical biosensor based on a Prussian-blue screen-printed electrode containing glucose oxidase incorporated into polyelectrolyte microcapsules (PMC) are considered. PMC with the embedded enzyme were formed using sodium polystyrene sulfonate and poly(allylamine hydrochloride). The characteristics were compared with those of the enzyme immobilized in chitosan gel. We assessed the dependences of biosensor signals on the composition of the buffer solution, on the glucose concentration; the operational and long-term stabilities. The enzyme immobilized in PMC proved to be more sensitive to buffer molarity at a maximum within 35 - 40 mM. The apparent Michaelis constants were 1.5 and 4.1 mM at the immobilization in, respectively, chitosan and PMC. The developed biosensors were used to assay commercial juices. The biosensors' data on the glucose contents were shown to have a high correlation with the standard spectrophotometric assay (0.92 - 0.95%), which implies a possible application of the fabricated biosensors in foodstuff analysis.