FF Farmacinių technologijų institutas (03.07)

This study represents a preliminary phytochemical and pharmacological screening of Anemone nemorosa L. collected in Ukraine. The aim was to characterize the phytochemical composition of the herb, rhizomes, and extracts obtained using solvents of different polarity, and to perform an initial evaluation of their toxicity, anti-inflammatory, hemostatic, and hepatoprotective activities. UPLC–MS/MS analysis identified 12 phenolic compounds, including flavonoids and hydroxycinnamic acids, with the highest level observed in AN-40 extract (obtained using 40% ethanol). Their content varies depending on the plant organ, location, and year of collection. For the first time, amino acid profiling identified 14 amino acids, complementing the phytochemical characterization of A. nemorosa herb and its extracts. The composition of phenolic compounds and amino acids depended on the extraction solvent. The highest total phenolic content was observed in AN-40 extract, whereas the highest flavonoid content was obtained with 70% ethanol. Toxicological evaluation demonstrated that A. nemorosa extracts may be classified as low-toxic substances (toxicity class IV), with no mortality observed at a dose of 5000 mg/kg. Preliminary pharmacological screening using single-dose evaluations in established experimental models indicated anti-inflammatory and hepatoprotective effects of A. nemorosa extracts. The extracts exhibited only weak hemostatic activity. Among the investigated extracts, AN-40 showed the most pronounced anti-inflammatory and hepatoprotective activities under the experimental conditions used. These preliminary findings warrant further dose–response, mechanistic, and bioactivity-guided studies.

Introduction Dermatophytosis caused by Microsporum canis is a common zoonotic skin infection in pets, often requiring prolonged treatment due to persistence in keratinized tissues. Its zoonotic nature highlights its importance within the One Health framework, as infected animals may contribute to transmission to humans. Organic acids represent promising antifungal agents due to their biocompatibility and mechanisms of action. Hydrogel systems may enhance topical drug delivery by enabling controlled release and improved retention. This study aimed to evaluate the antifungal activity of selected organic acids and to assess the physicochemical and antifungal properties of acid-based hydrogel formulations. Methods Ethanol-based solutions of gallic, coumaric, ferulic, and cinnamic acids (0.31–5%) were tested against M. canis using the agar well diffusion method. Cinnamic acid demonstrated the highest antifungal efficacy, which led to its selection for incorporation (0.5%) into hydrogel formulations containing poloxamer 407 and hydroxyethylcellulose with varying polymer ratios. Hydrogels were characterized by pH measurement and rheological analysis. Antifungal activity was evaluated using the same diffusion method. In vitro release studies were performed using a modified Franz diffusion cell system. Results Cinnamic acid demonstrated the highest antifungal activity, with inhibition zones increasing from 35.6 ± 1.7 mm (0.31%) to 73.2 ± 13.8 mm (5%). All hydrogel formulations retained antifungal activity against M. canis, producing ~17 mm inhibition zones, indicating diffusion limitations within the polymer matrix. Rheological analysis showed formulation-dependent viscosity (73.1–311.5 Pa·s at 35 °C), and all hydrogels had slightly acidic pH (4.81–4.92). In vitro, 34.8 ± 3.3% of cinnamic acid was released over 5 hours, suggesting a diffusion-controlled release profile. Conclusions Cinnamic acid demonstrated strong antifungal activity and retained its efficacy when incorporated into hydrogel systems, enabling controlled release. These findings indicate that cinnamic acid is a promising candidate for topical antifungal therapy, while poloxamer 407/hydroxyethylcellulose hydrogels may serve as a platform for prolonged local antifungal drug delivery

Background/Objectives: Primary non-adherence occurs when a prescribed medicine is never collected, preventing treatment from the beginning. Despite non-adherence being a widespread problem, most evidence comes from Western European and North American systems, and the Baltic region has rarely been assessed. Using Lithuania’s nationwide e-prescription system, we estimated primary non-adherence, identified associated factors, and characterised regional variation. Methods: We conducted a nationwide study of all outpatient electronic prescriptions issued in Lithuania, 2018–2024. We used open, anonymised quarterly prescribing and dispensing records from the national e-prescription subsystem and the national e-Health Services and Cooperation Infrastructure Information System (ESPBI IS). Prescribing and dispensing were deterministically linked. Prescriptions were classified as completed (dispensed quantity ≥ 90% of that prescribed), partially completed (dispensed quantity > 0% but <90%), or not completed (no quantity dispensed, defining primary non-adherence). Logistic regression estimated associations with patient, prescriber, prescription, and geographic characteristics; the results are reported as adjusted odds ratios (aOR) with 99% confidence intervals. Results: Of 108,204,007 prescriptions, 10.0% were never dispensed. Primary non-adherence was strongly associated with non-reimbursed status (aOR 3.47, 99% CI 3.46–3.48), younger age (18–44 years: 19.2% vs. ≥65 years: 7.0%), and short-term treatment (≤30 days: 14.1% vs. >90 days: 5.7%). Primary non-adherence rates were highest in cities (11.8%) and concentrated in the Vilnius metropolitan area (15.3%) and lowest in rural municipalities (5.5–6.0%). Conclusions: One in ten Lithuanian prescriptions is never dispensed. Non-adherence showed a marked, persistent geographic variance, with higher rates centred around Vilnius. Reimbursement status was the dominant correlate, pointing to cost as a key lever at the point of initiation.

Background: Three-dimensional printing (3DP) is rapidly advancing personalised medicine, yet systematic performance comparison with conventional manufacturing remains limited, particularly for plant-based formulations. Methods: This study compared tablets containing plant-based APIs (cannabidiol, apigenin, and luteolin) produced via conventional hot moulding and semi-solid extrusion (SSE) 3DP. The formulations were evaluated for physicochemical, mechanical, rheological, structural, and drug-release properties. Results: Both manufacturing methods produced tablets with comparable dimensions and mass; however, pronounced formulation-dependent differences were observed in mechanical strength, rheology, and microstructure. The molecular modelling predictions were consistent with the experimental findings. Agar–pectin exhibited the strongest predicted polymer–polymer and polymer–API interactions, including multiple hydrogen bonds, and formed a comparatively dense and cohesive matrix associated with slower API release. In contrast, the weaker interactions predicted for gelatine–pectin were associated with a less cohesive and more porous matrix that facilitated medium penetration, API diffusion, and drug release. SSE printlets generally exhibited greater porosity and more heterogeneous internal architectures than moulded tablets, resulting in enhanced drug release of approximately 95%. Micro-CT analysis provided important structural confirmation; API incorporation increased the void volume of gelatine–pectin printlets from 1.15% to 8.77%, demonstrating that disruption of polymer interactions contributed to pore formation and enhanced molecular diffusion. The observed release behaviour correlated with predicted molecular interactions and experimentally observed microstructural features, where increased porosity and weaker polymer–API interactions facilitated enhanced drug diffusion. Conclusions: Overall, SSE-3DP outperformed conventional moulding, demonstrating superior tunability and performance. This work provides a mechanistically informed strategy for designing plant-based, personalised natural products using 3DP technologies.

Single-needle electrospinning and needle-free electrospinning were applied to fabricate cellulose acetate (CA) fibrous mats containing rosemary (RO), clove (CL) and thyme (TH) essential oils (EO) from binary or ternary solvent mixture solutions. When an acetone, dichloromethane, and dimethylformamide mixture was used as the solvent system, cylindrical nanofibers with an average diameter between 255 and 617 nm were obtained. Removing dimethylformamide from the spinning solution resulted in flattened microfibers with significantly larger diameters, varying from 1242 to 2939 nm. Furthermore, CA nanofibers loaded with RO, CL or TH essential oils with diameter distributions ranging from 55 to 176 nm, from 69 to 215 nm and from 44 to 291 nm, respectively, were produced from ternary solvent mixture solutions using needle-free electrospinning equipment. FTIR and TGA studies revealed the incorporation of essential oils into the fiber structure, and GC-MS was used to evaluate the release of bioactive components from fibrous mats. The electrospun nanofibers of CA/RO, CA/CL and CA/TH showed excellent antioxidant activity and antimicrobial properties against Escherichia coli, Pseudomonas aeruginosa and Listeria monocytogenes. In addition, the CA/CL fibrous mat was tested in the storage of fresh beef steaks.

Rhododendron tomentosum Harmaja is an evergreen Ericaceae species recognized as a rich source of biologically active phenolic compounds. However, information on seasonal variation in its non-volatile phenolics throughout the annual phenological cycle remains limited. Therefore, this study investigated the influence of phenological development on the phenolic composition of R. tomentosum leaves collected from a natural population in Lithuania between October 2023 and October 2024. Phenolic profiles were determined using a validated HPLC-PDA method. A total of 25 phenolic compounds were identified, comprising phenolic acids, flavan-3-ols, procyanidins, flavonoids, and coumarin derivatives. The highest total phenolic content was observed during autumn-winter dormancy, whereas the lowest levels occurred during fruit development. Procyanidins and phenolic acids predominated during dormancy, while flavonoids, particularly quercitrin, hyperoside, and isoquercitrin, accumulated during flowering. PCA revealed clear phenological grouping of samples and demonstrated distinct associations between dormancy and procyanidin-rich profiles and flowering and flavonoid accumulation. These findings highlight phenological stage as a key factor influencing phenolic composition and indicate optimal harvesting periods depending on the target phenolic compounds.

Background: Transdermal films are promising dosage forms for controlled delivery of active pharmaceutical ingredients through the skin. Polyvinyl alcohol (PVA)-based matrices are particularly attractive due to their biocompatibility, film-forming ability, and versatility. However, the development of films for volatile compounds such as menthol remains challenging due to high potential losses during processing and application. This study compares PVA-based transdermal films fabricated by 3D printing, solvent casting, and electrospinning, focusing on the effect of fabrication method on film properties, release behaviour, and skin delivery performance.

Dried plums are valued as functional foods because of their nutritional composition and health-promoting properties. However, plum drying is challenging due to the waxy fruit cuticle, which limits moisture diffusion and prolongs drying time. Although blanching is commonly used to improve drying efficiency, its effects on phenolic compounds, minerals, and microbiological quality remain insufficiently studied, particularly in regional plum cultivars. Therefore, this study evaluated the effects of hot-water and 1% citric acid blanching combined with different drying methods on the nutritional, antioxidant, and microbiological quality of dried plums of the regional cultivar “Chuyskaya krasavitsa”. Dried plums contained 16–21 g of sorbitol, 26–42 g of mono- and disaccharides, 2–2.8 g of protein, 2.2–3.9 g of ash, and 2.9–4.5 mg of vitamin C per 100 g (wet basis). The predominant amino acids were aspartic acid, glutamine, and proline, while neochlorogenic acid, hyperoside, and chlorogenic acid were the major phenolic compounds. Dried plums were also rich in potassium (1294 mg/100 g on average), whereas freeze-dried samples retained higher mineral concentrations. Pretreatment significantly affected microbiological parameters, including aerobic colony count (p < 0.001) and yeast and mould count (p < 0.01), as well as protein, sugar, and total phenolic contents (p < 0.05). Citric acid blanching reduced the levels of several phenolic compounds compared with hot-water blanching and freeze-drying. These findings provide practical guidance for optimising dried plum processing while balancing nutrient retention, phenolic preservation, and microbial safety.

Introduction: Menthol, a naturally occurring terpene alcohol from Mentha piperita L., is incorporated into polyvinyl alcohol (PVA)-based pharmaceutical films for topical analgesia. Its high vapour pressure creates manufacturing challenges, inducing microstructural defects that compromise film mechanical performance. Characterizing the relationship between manufacturing-induced surface morphology and film mechanical properties is essential for rational biomaterial design in drug delivery applications. Methods: PVA-based films (PVA 10%, glycerol 3%) containing menthol (5% w/w) and benzocaine (5% w/w) were manufactured by three methods: semi-solid extrusion 3D printing, solvent casting, and DOBOT MG400 robotic arm-assisted electrospinning (15 kV, 1 mL/h, 10 cm tip-to-collector distance). Blank excipient-only controls were prepared in parallel. Surface microstructure was characterized by scanning electron microscopy (SEM) at x1,000 magnification. Mechanical properties were assessed by texture profile analysis: compression force, resilience, film burst strength (HDP/FSR probe), and rib seal strength (N/mm). The coefficient of variation (CV%) was calculated for all parameters. Results: SEM revealed distinct method-dependent microstructural profiles correlating directly with mechanical performance. Solvent casting produced large irregular surface pores (up to 50 um) from menthol volatility, corresponding to reduced burst strength (1202 ± 435 g, CV: 36.2%). 3D printing generated uniformly distributed controlled microporosity, yielding superior burst strength reproducibility (2369 ± 99 g, CV: 4.2%)-a 10-fold improvement over casting. Electrospun films preserved the intact nanofibrous architecture even with active compounds, demonstrating the highest compression resistance (37.5 ± 2.2 N, CV: 5.9%) and optimal rib seal strength (3.09 N/mm, CV: 7.7%). Menthol incorporation significantly reduced compression force in all methods compared to blank films, confirming matrix plasticization. Conclusions: Manufacturing method critically determines microstructural and mechanical quality of menthol-containing PVA biomaterial films for drug delivery. 3D printing provides superior dimensional reproducibility; electrospinning preserves nanofibrous integrity; and solvent casting is most vulnerable to volatile compound-induced defect formation. SEM morphology directly predicts mechanical performance, supporting its use as a quality screening tool in pharmaceutical biomaterial development.

Introduction: Menthol, a bioactive terpene alcohol from Mentha piperita L., activates epidermal TRPM8 cold receptors, so epidermal drug concentration is the most relevant pharmacological endpoint for topical analgesic applications. Semi-solid extrusion 3D printing enables precise layer-by-layer PVA matrix construction, potentially creating more homogeneous drug distribution than conventional solvent casting. Methods: PVA-based films (PVA 10%, glycerol 3%) containing menthol (5% w/w), benzocaine (5% w/w), and capsaicin (0-1% w/w) were manufactured by 3D printing (n=12) and solvent casting (n=12) using matched formulations. Ex vivo permeation studies used human abdominal skin from 10 donors in flow-through diffusion cells at 32C over 24 hours (ethics: Kaunas Regional Biomedical Research Ethics Committee, BE-2-42). Menthol was quantified by validated GC/FID (R2=0.99996). Statistics: Mann-Whitney U test; Cohen's d. Results: 3D printing produced a 4.4-fold higher epidermal menthol accumulation versus casting (22.0 vs 5.0 ug/mL; p=0.0004; Cohen's d=1.79). The epidermal-to-dermal ratio was markedly higher for 3D printing (0.230 vs 0.066), indicating preferential epidermal drug retention. Thickness uniformity CV was 7.3% for 3D printing versus 19.7% for casting; drug content RSD was 4.7% versus 9.5%. SEM of blank cast films showed smooth homogeneous surfaces, confirming that inferior precision in active formulations is attributable to menthol volatility during bulk drying rather than procedural deficiencies in the casting method itself. Conclusions: Taken together, these results suggest 3D printing warrants further investigation as a manufacturing platform for natural terpene-based transdermal biomaterials, offering superior epidermal drug retention and manufacturing reproducibility.