The Effect of Structural Characteristics of Deproteinized Spongy Bone on Activity of Adipose Tissue Mesenchymal Stromal Cells.

We studied the effect of structural properties of deproteinized spongy bone (DSB) on functional activity of adipose tissue mesenchymal stromal cells of (MSC) for the potential use of these materials as components of a combined tissue-engineered construct. The porosity of the structure of DSB samples and the pore size promote MSC adhesion, migration, and proliferation on their surface and in the depth, revealing the architectonics of this bone matrix. The depth of cell penetration into the samples (from 273 to 702 µm) and an increase in the total number of cells (from 302 on day 1 to 1744 on day 7) demonstrated MSC adhesion, migration, and proliferation. The viability of cultured MSC was preserved for up to 7 days. The obtained results prove the possibility of using allogeneic DSB from femoral heads as a bone matrix in tissue-engineered constructs in combination with MSC. Such constructs can be used to efficiently restore the structural and functional integrity of the bone tissue in abnormal processes of various etiopathogenesis associated with the formation of bone defects or bone tissue deficiency.

Structural Changes in the Lungs and Liver of Mice with Experimental Tuberculosis Treated with Liposome-Encapsulated Dextrazide.

Male BALB/с mice were intravenously infected with Mycobacterium tuberculosis H37Rv (0.5 ml of 2-week culture). One month later, treatment with liposome-encapsulated dextrazide (LEDZ, a conjugate of isonicotinic acid hydrazide (INH) and 40 kDa oxidized dextran encapsulated in phosphatidylcholine liposomes), INH, or a combination of LEDZ with INH was started. The doses of LEDZ (liposome suspension) and INH were 0.025 ml/10 g body weight and 5 mg/kg body weight, respectively. All the substances were administered 2 times a week via inhalation or intraperitoneally (a total of 40 doses). We studied the number and the size of tuberculous granulomas, the size of destruction foci and inflammatory infiltrates in the lungs and liver, the amount of fibrous connective tissue, and the dynamic of these parameters. LEDZ+INH inhalations were most effective by the therapeutic ratios in comparison with inhalation and intraperitoneal injections of INH.

Efficacy of Inhalations of Antituberculous Compositions with Different Length of Experimental Therapy Course in Mice.

The study compared antituberculous efficacy of individual or combined administration of "free" isoniazid and liposomal form of dextrazide (a composition consisted of isoniazid and oxidized dextran) inhaled in standard (15 mg/kg) or low (3 mg/kg) dose. The therapy started 1 month after contamination of outbred ICR male mice with Mycobacterium tuberculosis strain H37Rv. Combined inhalation of liposomal form of dextrazide and isoniazid in the low dose was most effective against mycobacterium tuberculosis due to diminished prodestructive pulmonary effect and a low hepatotoxicity. A minor prodestructive effect of this combination was observed starting from 1.5 month after the onset of therapy (12 inhalations, 2 times a week), and it augmented after 24 inhalations administered during 3 months.

[Effect of a gel based on recombinant human angiogenin on the healing of donor palate wounds]. / Vliianie gelia na osnove rekombinantnogo angiogenina cheloveka na zazhivlenie donorskikh ran tverdogo neba.

The aim of the study was to study the effect of the gel on the basis of recombinant human angiogenin on the rate of regeneration of donor palatal wounds. The study involved 20 patients (8 men and 12 women) aged 32 to 55 years. Patients were divided into two groups: the 1st group is a study group (n=10), whose patients in the postoperative period used a gel based on recombinant human angiogenin, the 2nd group is a control group (n=10) in which a gel based on recombinant human angiogenin was not used. Patients in both study groups underwent vestibuloplasty with simultaneous plasty of the attached keratinized gingiva with a free gingival graft from the area of the hard palate. The operations were carried out at the stage of disclosing dental implants, simultaneously with the installation of healing abatements or 4 weeks before dental implantation. For histological examination, tissue samples were obtained from the region of the edge of the donor's wounds of the palate at the 7th and 14th days after surgery. As a result of the study, significant differences were found in the comparison groups when assessing the processes of inflammation, angiogenesis and epithelization. The local application of the gel containing recombinant human angiogenin resulted in a rapid decrease in the intensity of inflammation in lamina propria mucosae and a significant decrease in the bulk density of cell infiltrates, accelerating regeneration. This is primarily due to the stimulation of the development of epidermal growth factor (EGF), vascular endothelial growth factor (VEGF) and increased blood supply to the affected area, as well as an increase in the proportion of fibroblasts. The most important observation was the increase in the rate of epithelialization of donor wounds of the hard palate.

Death Mechanisms of Pulmonary Alveolocytes in Mice Infected with Influenza Viruses A/H1N1/California/04/2009 and A/H5N1/Goose/Krasnoozerskoye/627/05.

In CBA mice infected with influenza viruses A/H1N1/California/04/2009 and A/H5N1/Goose/Krasnoozerskoye/627/05 in a dose of 10 MLD50, the mechanisms of death of pulmonary alveolocytes over 10 postinfection days were studied by light microscopy, immunohistochemistry, and morphometry. In mice infected with A/H1N1, alveolocytes died predominantly via necrosis, while apoptosis mostly employed the mitochondrial pathway. In mice infected with A/H5N1, apoptosis was the dominant mechanism of alveolocyte death proceeded via membrane receptor signaling followed by switching to FAS-mediated pathway via activation of FADD, the apoptotic signal transduction protein.

Changes in the Structure of Mouse Kidney in the Acute Period after Infection with Influenza Viruses A/H5N1 and A/H1N1.

Changes in the kidney structure in outbred and inbred male BALB/c mice were analyzed in the acute period after infection with influenza viruses A/H5N1 (10 MLD50; 10 days) and A/H1N1 (1 MLD50; 30 days). Antibodies to influenza viruses of both strains were most often expressed by endothelial cells of the glomeruli and arterioles and were rarely expressed by mesangiocytes and tubule epithelial cells. In the kidney, destructive processes induced by viruses and by ischemia due to massive blood vessel thrombosis. Mesangiocytes expressed factors, indicating that they could be qualified as M1 and M2 macrophages. Kidney destruction was more significant after infection of mice with the A/H5N1 virus, but in both experiments cell infiltrates were actually absent, probably due to blood vessel thrombosis and limited possibility of migration of mononuclear phagocytes and lymphocytes to the kidney.

Cell Death and Development of Fibrotic Alterations in Lung Granuloma of BALB/c Mice during Chronic BCG-Induced Granulomatosis.

Light microscopy, immunohistochemistry, and morphometric examinations established that cell death in lung granulomas of BCG-infected mice resulted mainly from activation of receptor-mediated apoptosis, which did not prevent the persistence of the causative agent in macrophages of the granulomas and promoted the formation of pronounced fibrosis in granulomas and pulmonary interstitium.

Preventive Effects of Oxidized Dextran on Functional Activity of Pulmonary Macrophages in Mice Infected with Influenza A Virus.

We analyzed cytokine profile of pulmonary macrophages in mice infected with highly pathogenic influenza A/H5N1 virus after preventive injections of oxidized dextran. Light microscopy, immunohistochemistry, and morphometric examinations showed that preventive injections of oxidized dextran led to more effective virus elimination, modulation of the proinflammatory cytokine response, and host antiviral response and reduce animal mortality. Our findings allow recommending oxidized dextran for further studies in order to create a vaccine with antiviral and adjuvant potencies.

Study of SMAD-Dependent Signal Pathway in the Development of Early Pulmonary Fibrosis in Mice Infected with Influenza A/H1N1 Virus.

Early fibrosis of the visceral organs is one of the main complications of infection caused by influenza A virus. Structural manifestations and molecular regulators of the epithelialmesenchymal transformation as a possible mechanism of fibrosis progression were studied in mice infected with influenza A/H1N1 A/Tomsk/13/2010 virus. We found early fibrosis of the lungs against the background of minor changes in fibroblast count. However, enhanced expression of TGF-ß and SMAD-2 by macrophages and alveolocytes attested to possible development of epithelial-mesenchymal transformation and its contribution to activation of fibrogenesis process in the lungs.

Studies of Influenza A/H1N1 A/Tomsk/13/2010 Virus Topology during Development of Infectious Process in Mammals.

Influenza A/H1N1 A/Tomsk/13/2010 virus registered in Siberia in 2010 proved to be an extremely pathogenic strain. Dynamic study of the topology of this influenza virus strain in the lungs, liver, kidneys, lymph nodes, and great vessels of infected mice was carried out. Influenza A virus was detected by immunohistochemical methods in cells of different histogenesis in all the studied organs throughout the observation period (days 1-30 postinfection), which indicated effective replication and long persistence of influenza A/H1N1 A/Tomsk/13/2010 virus in mammalian cells.

Fibrogenesis in granulomas and lung interstitium in tuberculous inflammation in mice.

The study in mouse model of BCG-induced granulomatous inflammation showed that early pulmonary fibrosis (day 3-30 postinfection) in tuberculous inflammation was primarily determined by increased number of fibroblasts in the lung interstitium and granulomas and enhanced fibroplastic activity. Fibroplastic processes are initiated via an increase in secretory activity of activated granuloma macrophages caused by the persistence of the pathogen in the cells of the mononuclear phagocytic system. The dynamics of hydroxyproline concentration under these conditions is determined by changes in the number and differentiation degree of fibroblasts in granulomas and lung interstitium at various stages of tuberculous inflammation.

Hydroxyproline content and fibrogenesis in mouse liver and lungs during the early stages of BCG granulomatosis.

In generalized BCG granulomatosis, fibrosis starts early (on day 3) and not only around the granulomas, but also in the organs. The severity of organ fibrosis is apparently determined by the concentration of granulomas, in particular their macrophages inducing proliferation of fibroblasts in organs and granulomas as well as activation of fibrogenesis. On day 30 after infection, the degree of fibrosis in the lungs was by 6 times higher than in the liver. The increase in hydroxyproline concentration in organs in early period of infection was determined by acute stress, while on day 30 it resulted from its enhanced synthesis by granuloma fibroblasts and resident fibroblasts in organs.

[The placental structural organization in pulmonary tuberculosis].

The placentas taken from 83 pregnant women with gestation of 37-40 weeks were morphologically and morphometrically studied. Group 1 comprised 23 pregnant women with pulmonary tuberculosis. Group 2 included 40 pregnant women whose pregnancy was complicated by gestosis. Group 3 (control) consisted of 20 healthy pregnant women. Group 1 was found to have chronic placental insufficiency caused by abnormal placental villous immaturity the basis of which was impaired processes of villous differentiation and terminal villous angiogenesis. In most cases, chronic placental insufficiency was compensatory in all the groups under study; but there were differences in compensatory mechanisms. The leading mechanism was cellular in Groups 1 and 2, the mixed (vascular and cellular) types of compensation was predominant in Group 3 (control).

Morphological changes in the liver of pregnant C57Bl/6 mice with BCG granulomatosis.

Morphological changes in the liver of pregnant C57Bl/6 mice infected with BCG vaccine before pregnancy and on day 13 of gestation were studied by means of light microscopy and morphometry. The formation of BCG granulomas in mice of both groups was followed by a decrease in the numerical density and diameter of granulomas, increase in the volume density of hepatocyte degeneration and necrosis, and rise in the numerical density of binucleate hepatocytes (compared to nonpregnant mice infected with BCG vaccine).

Structural changes in the liver of fetuses from female mice infected with BCG vaccine.

Hypotrophy was found in fetuses of female C57Bl/6 mice infected with BCG vaccine. Light microscopy of liver samples revealed destructive processes, impaired reparative regeneration, and fibrosis.

Morphological changes in the placenta of pregnant mice in experimental tuberculosis.

Changes in the placentas of pregnant C57Bl/6 mice infected with BCG vaccine were studied by light microscopy and morphometry. The decrease in the numbers of maternal and fetal vessels was paralleled by destructive changes and disorders in the compensatory reactions, which led to fetal hypotrophy after infection with BCG vaccine.