Usnea lichens are known for their beneficial pharmacological effects with potential applications in oral medicine. This study aims to investigate the extract of Usnea barbata (L.) Weber ex F.H. Wigg from the Călimani Mountains in canola oil as an oral pharmaceutical formulation. In the present work, bioadhesive oral films (F-UBO) with U. barbata extract in canola oil (UBO) were formulated, characterized, and evaluated, evidencing their pharmacological potential. The UBO-loaded films were analyzed using standard methods regarding physicochemical and pharmacotechnical characteristics to verify their suitability for topical administration on the oral mucosa. F-UBO suitability confirmation allowed for the investigation of antimicrobial and anticancer potential. The antimicrobial properties against Staphylococcus aureus ATCC 25923, Pseudomonas aeruginosa ATCC 27353, Candida albicans ATCC 10231, and Candida parapsilosis ATCC 22019 were evaluated by a resazurin-based 96-well plate microdilution method. The brine shrimp lethality assay (BSL assay) was the animal model cytotoxicity prescreen, followed by flow cytometry analyses on normal blood cells and oral epithelial squamous cell carcinoma CLS-354 cell line, determining cellular apoptosis, caspase-3/7 activity, nuclear condensation and lysosomal activity, oxidative stress, cell cycle, and cell proliferation. The results indicate that a UBO-loaded bioadhesive film’s weight is 63 ± 1.79 mg. It contains 315 µg UBO, has a pH = 6.97 ± 0.01, a disintegration time of 124 ± 3.67 s, and a bioadhesion time of 86 ± 4.12 min, being suitable for topical administration on the oral mucosa. F-UBO showed moderate dose-dependent inhibitory effects on the growth of both bacterial and fungal strains. Moreover, in CLS-354 tumor cells, F-UBO increased oxidative stress, diminished DNA synthesis, and induced cell cycle arrest in G0/G1. All these properties led to considering UBO-loaded bioadhesive oral films as a suitable phytotherapeutic formulation with potential application in oral infections and neoplasia.
The aim of our study was to obtain rutoside (RUT) inclusion complexes in β-cyclodextrin (β-CD) and in hydroxypropyl-β-cyclodextrin (HP-β-CD), in a 1:1 molar ratio, using the lyophilization method of complexation in solution. The complexes were confirmed and characterized, in comparison with the raw materials and their simple physical mixtures, by SEM, DSC, and FT-IR analyses. The antioxidant activity of the compounds was assessed by using the 2,2-diphenyl-1-picryl-hydrazyl (DPPH) and 2’-azino-bis(3-ethylbenzothiazolin-6-sulfonic) acid (ABTS) radicals, determining the radical scavenging activity, and by ferric reducing antioxidant power (FRAP) assay. The results revealed superior antioxidant ability for the inclusion complexes towards rutoside alone. The inclusion complexes were used as active ingredients in formulations of immediate-release tablets. The preformulation studies were performed on the powders for direct compression obtained after mixing the active ingredients with the excipients (Avicel PH 102, Polyplasdone XL-10, magnesium stearate, and talc). The materials were assessed for particle size, flowability, compressibility, and moisture content, establishing they are suitable for a direct compression process. The tablets were characterized regarding their pharmaco-technical properties and the results proved that the formulations lead to high-quality delivery systems, showing a good mechanical resistance with a low friability, excellent disintegration times, and satisfying dissolution rate. The performances were very similar for both formulations and the physico-mechanical properties of the tablets are not influenced by type of the used cyclodextrin, but the RUT- HP-β-CD tablets presented a higher dissolution rate.
Oxidative stress is associated with aging, cancers, and numerous metabolic and chronic disorders, and phenolic compounds are well known for their health-promoting role due to their free-radical scavenging activity. These phytochemicals could also exhibit pro-oxidant effects. Due to its bioactive phenolic secondary metabolites, Usnea barbata (L.) Weber ex. F.H. Wigg (U. barbata) displays anticancer and antioxidant activities and has been used as a phytomedicine for thousands of years. The present work aims to analyze the properties of U. barbata extract in canola oil (UBO). The UBO cytotoxicity on oral squamous cell carcinoma (OSCC) CLS-354 cell line and blood cell cultures was explored through complex flow cytometry analyses regarding apoptosis, reactive oxygen species (ROS) levels, the enzymatic activity of caspase 3/7, cell cycle, nuclear shrinkage (NS), autophagy (A), and synthesis of deoxyribonucleic acid (DNA). All these studies were concomitantly performed on canola oil (CNO) to evidence the interaction of lichen metabolites with the constituents of this green solvent used for extraction. The obtained data evidenced that UBO inhibited CLS-354 oral cancer cell proliferation through ROS generation (316.67 × 104), determining higher levels of nuclear shrinkage (40.12%), cell cycle arrest in G0/G1 (92.51%; G0 is the differentiation phase, while during G1 phase occurs preparation for cell division), DNA fragmentation (2.97%), and autophagy (62.98%) than in blood cells. At a substantially higher ROS level in blood cells (5250.00 × 104), the processes that lead to cell death—NS (30.05%), cell cycle arrest in G0/G1 (86.30%), DNA fragmentation (0.72%), and autophagy (39.37%)—are considerably lower than in CLS-354 oral cancer cells. Our work reveals the ROS-mediated anticancer potential of UBO through DNA damage and autophagy. Moreover, the present study suggests that UBO pharmacological potential could result from the synergism between lichen secondary metabolites and canola oil phytoconstituents.
Cereals whole grains contain vitamins, phytochemicals, antioxidants, resistant starch, and minerals with potential benefits to human health. The consumption of whole grains is correlated with a lowered risk of the most important chronic diseases, including type II diabetes, cardiovascular diseases, and some cancers. This study aimed to characterize and evaluate the content of five cultivars of wheat (Triticum aestivum L.) and five cultivars of barley (Hordeum vulgare L.) obtained by conventional plant breeding using crossing and selection methods. The novelty and the purpose of this research was to quantitatively and qualitatively analyze these ten cultivars from Romania and to show the importance of, and the changes produced by, crossing and selection methods when these are aimed at the physiological or morphological development of the cultivars. Studies based on gluten dosing; spectrophotometry using Bradford, fructan and protein dosing; Kjeldahl protein dosing; GC-MS/MS-protein and amino acid dosing; and identification of protein fractions using polyacrylamide gel electrophoretic method were conducted. This study demonstrates the possibility of developing future cultivars using conventional methods of improvement to modify the content and composition of nutrients to increase their health benefits.
The study is presenting the formulation and manufacturing process of 15 hydrogels formed by different cellulose derivative containing 1% naftifine hydrochloride dissolved in a mixture of cosolvents with low amounts of alcoholic components. The obtained products were evaluated by modelling their flow behaviour and by studying the in vitro release profile in vertical diffusion cell systems. The formulations showed a typical pseudoplastic behaviour. The range of variation in diffusion coefficient values is remarkably low, compared to structural differences easily detectable and demonstrated by rheological behaviour analysis. The increase in the concentration of the cellulose derivative matrix former leads to proportional decreases in diffusion coefficient values for naftifine, correlated with structural parameters. RezumatStudiul prezintă formularea și prepararea a 15 hidrogeluri generate de diferiți derivați de celuloză, conținând 1% clorhidrat de naftifină dizolvat într-un amestec de cosolvenți cu un conținut scăzut de componente alcoolice. Produsele obținute au fost evaluate prin modelarea comportamentului lor de curgere și prin studierea profilului de eliberare in vitro în sistemele celulare verticale de difuzie. Formulările au prezentat un comportament pseudoplastic tipic. Intervalul de variație a valorilor coeficientului de difuzie este foarte scăzut, comparativ cu diferențele structurale ușor detectabile și demonstrate prin analiza comportamentului reologic. Creșterea concentrației derivatului de celuloză duce la scăderea proporțională a valorilor coeficientului de difuzie pentru naftifină, corelată cu parametrii structurali.
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