The current trend in regenerative medicine, in the context of an aging population, is the search for new ways and means to optimize tissue bioengineering. One of the convenient models for in situ studying bone marrow regeneration is the subcutaneous ectopic osteogenesis test on scaffolds that imitate the architecture of bone tissue. Chelidonic acid (CA), a small molecule, is capable of participating in various cellular processes and metabolic pathways, and it can activate the osteogenic differentiation of mesenchymal stem cells. However, the molecular mechanisms behind the regulatory effects of CA remain unknown. The aim of this study was to investigate the modulatory effect of CA on the in situ formation of hematopoietic foci, as well as to predict target genes and intracellular signalling pathways involved in the hematopoietic activity of CA. An aqueous solution of CA, isolated from an extract of the Saussurea controversa plant. Course (daily for 35 days) oral administration of CA. Ectopic osteogenesis testing in Balb/c mice. Morphometric analysis of histological sections after 45 days and in silico modelling of gene expression with statistical analysis. CA, when administered orally in a low dose (10 mg/kg), threefold increases the normalized area of bone marrow in the composition of bone tissue plates grown in situ in a test of ectopic subcutaneous osteogenesis in mice. This effect is associated essentially (a probability of CA activity Pa 0.5 and a probability of inactivity Pi 0.5) with enhanced expression of 358 hematopoiesis-related genes, as predicted by in silico analysis. The top list with the highest Pa value included 10 target genes, such as GATA1, CITED2, SFRP1, EP300, LGALS9, VNN1, IL10RB, RARA, CD83, and HMOX1. CA has a significant ability to enhance the reparative remodelling of hematopoietic tissue in situ. The next phase of research will be to test actual target genes and signalling pathways that mediate the regulatory effect of HC on hematopoiesis both in vitro and in vivo, as well as in clinical settings.

The issue of delivery systems for various drugs and biological molecules, including scaffold technologies, is relevant, complex, and multifaceted, but is covered fragmentarily in the scientific literature. Many publications do not present the physicochemical properties of carrier materials or their biodegradation properties, which can affect the release of molecules from the matrix and their pharmacological activity. In other sources, the pharmacokinetics of drugs and/or cellular/tissue reactions are poorly described. As a result of this scattered information, it is difficult to search for materials in a targeted matter and unambiguous conclusions cannot be drawn on topical issues. Based on review and original articles, information on the development and functioning of scaffolds as carriers of medicinal and biological molecules is collected and critically examined. Materials and substances used in delivery systems of drugs and biological molecules, as well as cellular and tissue reactions during their use, are classified. Particular attention in the review is given to composite scaffolds with a calcium phosphate component as carriers of various pharmacological agents for effective delivery systems in bone-tissue bioengineering.

T-lymphoblast-like human leukemia cells of the Jurkat line (Jurkat T cells) form polyploid forms with a high DNA content in suspension culture. Due to pronounced genetic instability, this contributes to further transformation and development of clonal diversity (polyclonal) of the cell line. Little information is available on the adherent subpopulation of Jurkat T cells. In this work, we analyzed the content of nucleic acids in suspension (DNA) and adhesive (DNA, RNA) subpopulations of Jurkat T cells using flow cytometry and propidium iodide dye, as well as confocal laser microscopy and acridine orange dye. The morphology and mobility of large (with a diameter of more than 15 μm) Jurkat T cells adhering to plastic were studied using Cell-IQ phase-contrast microscopy in real-time. According to the intensity of fluorescence in the conditionally green wavelength range (300–530 nm: from UV to green) and conditionally red (565–800 nm: from red to far-red), three subpopulations of adherent Jurkat T cells were identified: with high, medium, and low nucleic acid content. Thus, Jurkat-T cells adhering to the plastic surface of the plates retain a pronounced heterogeneity in the DNA content characteristic of the suspension fraction, which suggests a difference in the morphofunctional properties (polyclonicity) of this subpopulation of cell culture. With a sharp increase in the total cell mass, the proportion of large (giant, 15–50 μm or more) cells attached to the plastic remained constant for 21 days of cultivation and amounted to 1% of the adhesive fraction. It was found that large Jurkat T cells (with a median diameter of 31 µm) moved along the plastic at a linear (median) speed of 38 µm/h. The polynuclear Jurkat T cells on plastic are morphologically identified, with a linear increase being revealed in the mobility of adherent cells with an increase in their diameter (regression coefficient r = 0.33, p < 0.02, n = 52). Possible cellular and molecular mechanisms of an increased number of DNA copies in some adhering Jurkat T cells are discussed. It is assumed that the discovered new property (locomotor activity) can provide polyploid (multinucleated) adhering Jurkat T cells with a significant advantage—directed migration (chemotaxis) in a growing cell population under conditions of nutrient deficiency due to a change in the nutrient medium after 3–4 days of cultivation.

Курсовое применение экстрактов Saussurea controversa DC (S.c.) и Fillipendula ulmaria (L). Maxim. (F.u.), содержащих широкий набор флавонолгликозидов, при экспериментальном остеомиелите приводит к стимуляции гранулопоэза в 2,8 и 2,2 раза соответственно (p = 0,04) и регенерации костной ткани (плотность распределения остеобластов возрастает в 5 раз, p = 0,02) в зоне поражения у крыс. В результате исследования миелограмм костного мозга отмечены активация митотической активности гранулоцитов, увеличение количества миелобластов в 5,6 и 4,2 раза (p = 0,02) и зрелых форм гранулоцитов (в 2,5 и 1,5 раза, p = 0,04), измененных в результате антибиотикотерапии. После применения экстракта S.c. в костном мозге крыс с остеомиелитом возрастает общее количество ядросодержащих клеток в 2,7 (p = 0,04) раза. Улучшение морфологической картины костной ткани (активация клеток эндоста, периоста, появление остеобластических «почек», формирование грануляционной ткани) и формирование зрелой костной ткани более выражено (на 12 %, p = 0,02) при применении экстракта S.c., по сравнению с экстрактом F.u.

The aim of the work was to study the effect of fractions from Saussurea controversa and Мeadowsweet extracts on hemopoiesis and regeneration of bone tissue, which are the main target indicators in experimental osteomyelitis.Materials and methods. In the experiment we used butanol, water and element-organic fractions from the Saussurea controversa extract as well as ethyl acetate, butanol and water fractions from the Мeadowsweet extract, obtained by liquid extraction of the extracts from corresponding plants with the help of organic solvents. After modeling osteomyelitis of the right femur in rats and course treatment for 28 days, the state of bone marrow hemopoiesis was evaluated and morphological examination of the affected limb was performed.Results. The butanol fraction (at a dose of 10 mg/kg) containing flavonol glycosides, the water fraction (at a dose of 80 mg/kg) containing polysaccharides in its composition and the element-organic fraction (at a dose of 10 mg/kg) represented by organic calcium showed the highest osteogenic activity in the Saussurea controversa extract. The water fraction from the Meadowsweet extract (at a dose of 50 mg/kg) demonstrated pronounced hemopoietic activity due to stimulating erythro-, granulo- and lymphopoiesis in the bone marrow of rats with experimental osteomyelitis.