Development and Evaluation of Two-Phase Gel Formulations for Enhanced Delivery of Active Ingredients: Sodium Diclofenac and Camphor.

The formulation of biphasic gels as potential semi-solid carriers for hydrophilic and lipophilic active substances is promising for the development of pharmaceutical preparations. The aim of this study was to design a stable bigel composition and to determine the influence of the organogel/hydrogel ratio on the gel's physical-chemical and structural-mechanical properties. The investigated compositions of organogel/hydrogel remained stable at ratios ranging from 5/95 to 40/60. After texture and microstructure analysis, bigels with 20/80 and 25/75 ratios were selected as carriers for the active ingredients, sodium diclofenac and camphor, for use as topical preparations for the treatment of muscle-joint inflammation and pain. Although other researchers have published data on the preparation and evaluation of bigels, there are no scientific results on the development of a two-phase gel with our proposed combination of APIs. Sodium diclofenac release was found to be higher when combined with camphor, which revealed the advantages of the biphasic formulation. The pseudoplastic behavior, thixotropy, and thermal stability of flow of the studied bigel samples was investigated by rheological analysis. Ongoing stability studies confirmed the minimal 6-month period.

The Study of the Cytotoxicity, Proliferative and Microbiological Activity of the Medicated Chewing Gum with Ascorbic Acid and Lysozyme Hydrochloride Using Different Culture of Cells.

Medicated chewing gum with lysozyme hydrochloride and ascorbic acid as active pharmaceutical ingredients was developed for application in dentistry. The aim of this research was to study the cytotoxicity, proliferative, and microbiological activities of the active ingredients in different types of cell cultures. The preclinical study of active pharmaceutical ingredients and their combinations was carried out using culture lines such as HepG2 (human hepatocarcinoma cells), Hek293 (human embryonic kidney cells), and MAEC (mouse aortic endothelial cells). MTT assays were used to analyse cytotoxicity and proliferative activity, while the state of antioxidant protection was assessed by the content of sulfhydryl groups and catalase activity. The determination of lipid peroxidation products was based on the level of TBA-active products. As a microbiological model for studying the effect of the developed dental medicine on the ability of the oral cavity microorganisms to form biofilms, the following strains were used: Streptococcus mutans, Staphylococcus aureus, Staphylococcus epidermidis, Lactobacillus plantarum, and Candida albicans. The optical density of the formed biofilm was evaluated by the intensity of the experimental sample's colour on a StatFax 303 Plus photometer at a wavelength of 630 nm. The combination of ascorbic acid and lysozyme hydrochloride in the established concentrations (20 mg and 10 mg per 1 gum, respectively) resulted in a slight stimulation of cell proliferation without any toxic effects and increased antioxidant protection, preventing the development of oxidative stress. It was found that, in contrast to the separately used active substances, the combination of lysozyme hydrochloride and ascorbic acid inhibits the biofilm formation of all studied microorganisms and shows the ability to destroy diurnal biofilms of L. plantarum and fungi of the genus Candida, indicating potentiation and summation of the active pharmaceutical ingredients' composition effects in the developed dental medicine. Due to the observed positive pharmacological and microbiological action, the combination of lysozyme hydrochloride and ascorbic acid in the medicated chewing gum serves as a promising tool for the prevention and treatment of infectious and inflammatory diseases of the periodontium and mucous membranes and the prevention of caries.

Impact of Compression Force on Mechanical, Textural, Release and Chewing Perception Properties of Compressible Medicated Chewing Gums.

Medicated chewing gums (MCGs) represent a beneficial platform for realizing drugs intended for dental prophylaxis and treatment. The present study aimed to investigate the impact of compression force on the mechanical, textural, release, and chewing perception characteristics of compressible MCGs with the combination of lysozyme hydrochloride (LH) and ascorbic acid (AsA). Four batches of MCGs were obtained on a laboratory single-punch tablet machine applying different forces, i.e., 5, 7, 10, and 15 kN, and evaluated by their geometrical parameters, mechanical resistance, surface and internal structure characteristics, texture profile, release behavior, and perception attributes during mastication. It was found that increasing compression force slightly affected resistance to crushing and friability of MCGs, but resulted in surface smoothing and formation of a thicker layer with highly compacted particle arrangement. According to the texture analysis, increasing compression force led to harder and more adhesive gums, indicating possible difficulties in chewing and, therefore, impairment of their consumer properties. Lower compression forces were also found to be preferable in terms of better drug release from the obtained chewing gums. The volunteers' assessment showed that an increase of compression force led to significantly raising the initial hardness and crumbliness as well as to decreasing the rate of the integral gum mass formation during mastication, which may negatively affect perceptive sensations when using MCGs. Based on the results obtained, the optimal compressing force was selected to be 7 kN, which allows obtaining MCGs with good organoleptic, mechanical, textural, and release properties.

Determination of Organic Compounds, Fulvic Acid, Humic Acid, and Humin in Peat and Sapropel Alkaline Extracts.

Black, brown, and light peat and sapropel were analyzed as natural sources of organic and humic substances. These specific substances are applicable in industry, agriculture, the environment, and biomedicine with well-known and novel approaches. Analysis of the organic compounds fulvic acid, humic acid, and humin in different peat and sapropel extracts from Lithuania was performed in this study. The dominant organic compound was bis(tert-butyldimethylsilyl) carbonate, which varied from 6.90% to 25.68% in peat extracts. The highest mass fraction of malonic acid amide was in the sapropel extract; it varied from 12.44% to 26.84%. Significant amounts of acetohydroxamic, lactic, and glycolic acid derivatives were identified in peat and sapropel extracts. Comparing the two extraction methods, it was concluded that active maceration was more efficient than ultrasound extraction in yielding higher amounts of organic compounds. The highest amounts of fulvic acid (1%) and humic acid and humin (15.3%) were determined in pure brown peat samples. This research on humic substances is useful to characterize the peat of different origins, to develop possible aspects of standardization, and to describe potential of the chemical constituents.

The Influence of pH Values on the Rheological, Textural and Release Properties of Carbomer Polacril® 40P-Based Dental Gel Formulation with Plant-Derived and Synthetic Active Components.

The physicochemical properties, especially pH value of dental medicines, have significant influence on the health of oral cavity tissues. The pH of formulations should correspond to the value of saliva pH (5.5-8.0). For carbomer-based gels, the required pH value is obtained by neutralizing them with alkaline components, which leads to their structuring (thickening). This affects the physical properties of the gel, its residence time at the application site and the rate of release of active pharmaceutical ingredient. Therefore, the main purpose of this study is to evaluate the rheological, textural, and biopharmaceutical properties of Carbomer Polacril® 40P-based dental gel depending on the pH value. Evaluation of the rheological properties of gel preparations were performed by measuring the structural viscosity of the samples as a function of pH and temperature. The textural properties of the gel were evaluated by performing tests regarding back extrusion and spreadability. Carbomer Polacril® 40P-based gels haven't shown noticeable thixotropic behavior, and were characterized by plastic flow in the whole studied pH range. The structural viscosity at the selected average pH value hasn't differed at storage (25 °C) and application (37 °C) temperature. Texture studies of dental gels have shown a strong correlation with rheoparameters. Their rheological behavior and textural properties haven't changed significantly between the pH range of 5.5-6.6. The relatively narrow range of working pH values does not affect the change in the viscosity of the preparation significantly and, consequently, does not affect the release of APIs from the developed Carbomer Polacril® 40P-based dental gel.

Development of New Formula Microcapsules from Nutmeg Essential Oil Using Sucrose Esters and Magnesium Aluminometasilicate.

Essential oils are volatile liquids which evaporate and lose their pharmacological effect when exposed to the environment. The aim of this study is to protect nutmeg essential oil from environmental factors by encapsulation (shell material, sodium alginate) and determine the influence of crosslinker concentration (2%, 5% calcium chloride), different emulsifiers (polysorbate 80, sucrose esters), and magnesium aluminometasilicate on microcapsule physical parameters, encapsulation efficiency (EE), swelling index (SI), and other parameters. Nutmeg essential oil (NEO)-loaded calcium alginate microcapsules were prepared by extrusion. The swelling test was performed with and without enzymes in simulated gastric, intestinal, and gastrointestinal media. This study shows that the crosslinker concentration has a significant influence on EE, with 2% calcium chloride solution being more effective than 5%, and capsules being softer with 2% crosslinker solution. Using sucrose esters, EE is higher when polysorbate 80 is used. The swelling index is nearly three times higher in an intestinal medium without enzymes than in the medium with pancreatin. Microcapsule physical parameters depend on the excipients: the hardest capsules were obtained with the biggest amount of sodium alginate; the largest with magnesium aluminometasilicate. Sucrose esters and magnesium aluminometasilicate are new materials used in extrusion.

Natural Ingredients-Based Gummy Bear Composition Designed According to Texture Analysis and Sensory Evaluation In Vivo.

The increased interest in functional materials of natural origin has resulted in a higher market demand for preservative-free, "clean label", or natural ingredients-based products. The gummy bear food supplements are more acceptable to consumers and have fewer limitations compared to other dosage forms. The aim of our study was to produce natural ingredients-based gummy bear composition, and evaluate the influence of the selected ingredients on the product's textural properties, its acceptance in vivo, and the gummy bear's quality. The optimal base composition was determined using a surface response design: gelatin 4.3 g and agave syrup 6.3 g. The investigated sweeteners did not affect the textural properties (p > 0.05). However, further studies demonstrated that a 100% increase of agave results in up to 27% higher flexibility (p < 0.05). The addition of calcium and cholecalciferol reduced firmness by 59.59 ± 1.45% (p < 0.05). On the other hand, acai berry extract had no significant effect. The presence of calcium resulted in a decreased smell and taste; however, the data indicated that experimental texture analysis is a more accurate technique than in vivo evaluation. The acai berry extract did not improve all of the tested sensory properties. We can conclude that the suggested gummy bear base can be supplemented with various active ingredients and commercialized, though further studies are needed to investigate the other natural sources to mask the unpleasant taste of active ingredients and avoid water loss.

Formulation and Characterization of Potential Antifungal Oleogel with Essential Oil of Thyme.

The aim of this research was to formulate oleogel with thyme essential oil with potential antimicrobial activity, design optimal formulation, and evaluate the influence of ingredients on texture parameters of preparation. Central composite design was applied to statistical optimization of colloidal silica and paraffin oil mixture for the modeling of oleogel delivery system. The influence of designed formulations on response variables (texture parameters), firmness, cohesiveness, consistency, and index of viscosity, was evaluated. Quality of essential oil of thyme was assessed by determinate concentration of thymol and carvacrol using gas chromatography with flame ionization detection (GC-FID). Microbiological tests have shown that oleogel with thyme essential oil affects Candida albicans microorganism when thyme essential oil's concentration is 0,05% in oleogel mixture.

Evaluation of Beeswax Influence on Physical Properties of Lipstick Using Instrumental and Sensory Methods.

The aim of this study was to optimize the lipsticks formulation according to the physical properties and sensory attributes and investigate the relationship between instrumental and sensory analyses and evaluate the influence of the main ingredients, beeswax and oil, with analysis of lipsticks properties. Central composite design was used to optimize the mixture of oils and beeswax and cocoa butter for formulation of lipsticks. Antioxidant activity was evaluated by DPPH free radical scavenging method spectrophotometrically. Physical properties of lipsticks melting point were determined in a glass tube; the hardness was investigated with texture analyzer. Sensory analysis was performed with untrained volunteers. The optimized mixture of sea buckthorn oil and grapeseed oil mixture ratio 13.96 : 6.18 showed the highest antioxidative activity (70 ± 0.84%) and was chosen for lipstick formulation. According to the sensory and instrumental analysis results, optimal ingredients amounts for the lipstick were calculated: 57.67% mixture of oils, 19.58% beeswax, and 22.75% cocoa butter. Experimentally designed and optimized lipstick formulation had good physical properties and high scored sensory evaluation. Correlation analysis showed a significant relationship between sensory and instrumental evaluations.

Alternative preparation of propolis extracts: comparison of their composition and biological activities.

BACKGROUND: Propolis is the bee product noted for multiple biological effects, and therefore it is widely used for the prevention and treatment of a variety of diseases. The active substances of propolis are easily soluble in ethanol. However ethanolic extracts cannot be used in treatment of certain diseases encountered in ophthalmology, pediatrics, etc. Unfortunately, the main biologically active substances of propolis are scarcely soluble in water, oil and other solvents usually used in pharmaceutical industry. The aim of this study was to investigate chemical composition, radical scavenging and antimicrobial activity of propolis extracts differently made in nonethanolic solvents. METHODS: Total content of phenolic compounds in extracts was determined using Folin-Ciocalteu method. Chemical composition and radical scavenging activity of extracts were determined using HPLC system with free radical reaction detector. Antimicrobial activity of examined preparations was evaluated using the agar-well diffusion assay. RESULTS: Total amount of phenolic compounds in extracts made in polyethylene glycol 400 (PEG) and water mixture or in PEG, olive oil and water mixture at 70 °C was comparable to that of ethanolic extract. Predominantly identified compounds were phenolic acids, which contribute ca. 40 % of total radical scavenging activity. Investigated nonethanolic extracts inhibited the growth and reproduction of all tested microrganisms. Antimicrobial activity of some extracts was equal or exceeded the antimicrobial effect of ethanolic extract. Extracts made in pure water or oil only at room temperature, contained more than 5 - 10-fold lower amount of phenolic compounds, and demonstrated no antimicrobial activity. CONCLUSIONS: Nonethanolic solvent complex and the effect of higher temperature allows more effective extraction of active compounds from propolis. Concentration of total phenolic compounds in these extracts does not differ significantly from the concentration found in ethanolic extract. Propolis nonethanolic extracts have radical scavenging and antimicrobial activity.

Analysis of content of phenolic acids in Lithuanian propolis using high-performance liquid chromatography technique.

UNLABELLED: The aim of the study was to analyze phenolic acids in Lithuanian propolis and to compare it with the composition of propolis in neighboring countries (Latvia and Poland) according to the predominant flora in the collecting places. The study was also aimed at the evaluation of the effect of the layer thickness (mm) of the harvested propolis on the quality of the raw material in determining the amount of phenolic acids. MATERIALS AND METHODS: The object of the study was propolis collected in Lithuania, Poland, and Latvia in late July of 2006 and 2007. The qualitative and quantitative analysis of phenolic acids was performed using the high-performance liquid chromatography technique (HPLC). RESULTS: The results of the study showed that the quantitative and qualitative composition of phenolic acids in propolis depended on the plants from which the bees in the area collected substances for the raw material of propolis. The predominant phenolic acids were determined to be ferulic and coumaric acids, and they may be among the main indicators of quality in the standardization of the raw material and preparations of propolis. CONCLUSION: We created an HPLC-based analysis method for the identification and quantification of phenolic acids in propolis. The variety of phenolic acids in propolis depends on the vegetation predominating in the harvesting area. Studies have shown that the highest amount of phenolic acids is observed in propolis harvested in areas characterized by the predominance of deciduous trees and meadows. Results have also shown that ferulic and coumaric acids are the predominant phenolic acids in propolis. The thickness of the layer of the collected propolis in the hive also influences its chemical composition.

The comparison of anti-oxidative kinetics in vitro of the fluid extract from maidenhair tree, motherwort and hawthorn.

The aim of the study was to perform a quantitative analysis of fluid extracts of maidenhair tree (Ginkgo biloba L.), motherwort (Leonurus cardiaca L.) and hawthorn (Crataegus monogyna Jacq.), to evaluate their antioxidant activity and to compare their ability to inactivate free radicals. The antioxidant activity was measured using the DPPH*and the ABTS*+ radical scavenging reaction systems. The study showed that the manifestation of the radical scavenging capacity in the DPPH* reaction system was in the following order: the fluid extract of hawthorn (70.37 +/- 0.80%) > the fluid extract of maidenhair tree (82.63 +/- 0.23%) > the fluid extract of motherwort (84.89 +/- 0.18%), while in the ABTS*+ reaction system, the manifestation of the radical scavenging capacity was in the following order: the fluid extract of hawthorn (87.09 +/- 0.55%) > the fluid extract of motherwort (88.28 +/- 1.06%) > the fluid extract of maidenhair tree (88.39 +/- 0.72%). The results showed that in the DPPH* reaction system, fluid extract of motherwort manifested higher antioxidant activity, compared to the fluid extracts of maidenhair tree and hawthorn. By contrast, in the ABTS*+ reaction system, higher antioxidant activity was found in the fluid extract of maidenhair tree, compared to the fluid extracts of motherwort and hawthorn. This would suggest that preparations manufactured from these herbal raw materials could be used as effective preventive means and valuable additional remedies in the treatment of diseases caused by oxidative stress.

Pooling of urine specimens allows accurate and cost-effective genetic detection of Chlamydia trachomatis in Lithuania and other low-resource countries.

The aims of this study were to compare performance characteristics and cost-effectiveness of pooling urine samples for screening and diagnosis of Chlamydia trachomatis using Digene Hybrid Capture II CT/NG Test (HCII), and to examine the prevalence of C. trachomatis in male military recruits in Lithuania. A total of 410 urine samples were individually tested and pooled by 5 and 10 samples, respectively. The sensitivity and specificity of diagnosis were not affected by either pooling strategy. The estimated population prevalence of C. trachomatis infection was nearly identical, i.e. 4.4%, 4.4% and 4.1% based on individually tested samples, and samples pooled by 5 and 10, respectively. For this estimation of the population prevalence, pooling 5 samples reduced the costs by 80% and pooling 10 samples reduced the costs by 90%. For diagnosis of each individual sample, the pooling strategies resulted in cost savings of 60% (5 samples per pool) and 56% (10 samples per pool). The present pooling strategies were sensitive, specific and cost-efficient for screening and diagnosis of C. trachomatis infection in male military recruits in Lithuania. The strategies would be most useful for reasonably inexpensive large-scale screening, prevalence studies and even diagnostics in Lithuania and many other low-resource countries.

[Influence of sea buckthorn oil production technology on its antioxidant activity]. / Saltalankiu aliejaus technologijos itaka jo antioksidaciniam aktyvumui.

The aim of the study was to determine quality and quantity of the main components of sea buckthorn oils, which were produced by different technological procedures, and to evaluate their antioxidant activity. An influence of thermal shock to antioxidant activity of sea buckthorn oils was also evaluated. Two kinds of sea buckthorn oils were chosen. Oil of pulp was produced by separating oil from soft part of the berries. The other oil was produced using berries extracted with fatty oils. The content of carotenoids (by spectrophotometry at 450 nm) and relative content of fatty acids (gas chromatography) were determined in sea buckthorn oils. Antioxidant activity was evaluated for the first moment and after storing in a thermostat at +60+/-2 degrees C temperature for 1, 2 and 8 hours. 2,2-diphenyl-1-picrylhydrazyl (DPPH*) and 2,2'-azino-bis-(3-ethilbenzthiazolin-6-sulphonic acid) (ABTS*+) methods were used. The results have demonstrated that extracted oil has 2.4 times more carotenoids. Analysis of sea buckthorn oils proved a difference of content of the fatty acids. Extracted oil is more potent antioxidant, showed by both methods. After storing samples at +60+/-2 degrees C temperature, antioxidant activity decreased. The results proved that the oils obtained by different production procedures have different content and different antioxidant activity.

[Stability and antioxidant activity of black currant and black aronia berry juices]. / Juoduju serbentu ir juodavaisiu aroniju uogu sulciu stabilumas ir antioksidacinis aktyvumas.

The berries of black currant and black aronia are rich in polyphenolic compounds and especially in anthocyanins, demonstrating antioxidant activity. The aim of the study was to evaluate the possible effect of thermal technological processes on the quantity of polyphenols and anthocyanins in berry juice concentrates, and on the antioxidant activity. After 8 hour storage of black currant and black aronia berry juice concentrates at 60 degrees C, the amount of polyphenols decreased by 46% and 22%, anthocyanins 31% and 35%, respectively. Antioxidant activity decreased by 26% and 56%, respectively. The results demonstrated insufficient stability of juice concentrates, and impropriety of application of long lasting drying processes in manufacturing of black currant and black aronia berry dry products. Fast and efficient drying methods for liquid products should be applied to preserve qualitative and quantitative composition and their antioxidant activity.