Bone grafts with a high potential for osseointegration, capable of providing a complete and effective regeneration of bone tissue, remain an urgent and unresolved issue. The presented work proposes an approach to develop composite biomimetic bone material for reconstructive surgery by deposition (remineralization) on the surface of high-purity, demineralized bone collagen matrix calcium phosphate layers. Histological and elemental analysis have shown reproduction of the bone tissue matrix architectonics, and a high-purity degree of the obtained collagen scaffolds; the cell culture and confocal microscopy have demonstrated a high biocompatibility of the materials obtained. Adsorption spectroscopy, scanning electron microscopy, microcomputed tomography (microCT) and infrared spectroscopy, and X-ray diffraction have proven the efficiency of the deposition of calcium phosphates on the surface of bone collagen scaffolds. Cell culture and confocal microscopy methods have shown high biocompatibility of both demineralized and remineralized bone matrices. In the model of heterotopic implantation in rats, at the term of seven weeks, an intensive intratrabecular infiltration of calcium phosphate precipitates, and a pronounced synthetic activity of osteoblast remodeling and rebuilding implanted materials, were revealed in remineralized bone collagen matrices in contrast to demineralized ones. Thus, remineralization of highly purified demineralized bone matrices significantly enhanced their osteostimulating ability. The data obtained are of interest for the creation of new highly effective osteoplastic materials for bone tissue regeneration and augmentation.
Due to the small number of studies on the role of coumarins in associative symbiotic relationships, some aspects of the influence of synthetic coumarins on the physicochemical and cultural properties of Azospirillum baldaniorum Sp245 were studied for the first time. To reveal the role of hydroxylation in position 7 of the fused aromatic ring – 1-(2-oxo-2H-chromen-3-yl)butan-1,3-dione, comparative studies of the effect of the original and hydroxylated coumarins on the culture of a model strain of azospirilla were carried out. The survival of bacteria after the addition of coumarins was studied by counting CFU on an agar medium. The biofilm formation activity of the culture was assessed using crystal violet. The change in the surface of bacteria under the action of coumarins was studied by the electrical polarizability of bacterial cells on an ELUS electrooptical analyzer (EloSystemGbR, Germany). The yield and monosaccharide composition of extracellular glycopolymers were studied using gas-liquid chromatography.For the first time, an increase in the yield of EPS of bacteria during growth in the presence of 1-(7-hydroxy-2-oxo-2Hchromen-3-yl)butan-1,3-dione by 1.2 and 1.7 times for concentrations of 50 and 100 μM respectively was observed. It has been established for the first time that the hydroxylated substance has a higher antibacterial activity compared to the unsubstituted one. A decrease in the number of viable cells in planktonic culture and inhibition of biofilm growth were revealed. It has been shown by electro-optical analysis that the presence of coumarins in the cultivation medium in all concentrations studied leads to a change in the electrical polarizability of A. baldaniorum Sp245 cells. The use of electrooptical analysis of cell suspensions using monospecific antibodies obtained against lipopolysaccharides of this strain made it possible to reveal the absence of changes in carbohydrate antigenic determinants on the surface of bacterial cells. This is consistent with the data of the analysis of the composition of extracellular polysaccharides by GLC, during which no differences were found in the qualitative composition and ratio of monosaccharides. An increase in the yield of bacterial EPS during growth in the presence of 1-(7-hydroxy-2-oxo-2H-chromen-3-yl) butan-1,3-dione by 1.2 and 1.7 times for concentrations of 50 and 100 μM was shown. The results obtained allow us to consider the changes that have occurred as features of the adaptation of bacteria to the associative conditions of existence.
The possibility of detection and determination of flavonoids by using microbial cells was shown for the first time using the quercetin - Azospirillum baldaniorum Sp245 model system. The activity of the flavonoids quercetin, rutin and naringenin toward A. baldaniorum Sp245 was evaluated. It was found that when the quercetin concentration ranged from 50 to 100 µM, the number of bacterial cells decreased. Rutin and naringenin did not affect bacterial numbers. Quercetin at 100 μM increased bacterial impedance by 60 %. Under the effect of quercetin, the magnitude of the electro-optical signal from cells decreased by 75 %, as compared with the no-quercetin control. Our data show the possibility of developing sensor-based systems for the detection and determination of flavonoids.
This article discusses the prospects for the use of new elastin barrier membranes manufactured using adapted technologies for the selective isolation of the elastin component from the extracellular xenogenic matrix of the pericardium ligamentous apparatus: (1) by high-temperature extraction under pressure; (2) cyanogen bromide method. A commercial material, Geistlich Bio–Gide® membrane (BG), was used as a control comparison group. It is shown that the materials of group (1) have a high degree of biocompatibility, exceeding the indicators of the control group BG. Based on the results of an study in a model of subcutaneous heterotopic implantation in rats, it was shown that elastin BM has a chemoattractant effect on the mesenchymal recipient cells and, unlike the control, is able to integrate to a high degree into the surrounding recipient tissues. At the same time, the materials of group (1) had a pronounced proangiogenic effect. Thus, it has been shown that elastin BM groups (1) have a medium-term barrier function and are able to induce full-fledged cellular repopulation and local neoangiogenesis, which can be useful in clinical practice, primarily in GTR technologies (with gingival flap augmentation) or when used together with other BM as an angiogenesis inducer to ensure formation of the vascular bed in GBR technologies of bone tissue.
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