Possible Kidney Complications after Application of Cell Technologies for the Repair of the Resected Liver.

The state of rat kidneys after injection of bone marrow multipotent stromal cells (MSC) labeled with Vybrant CM-Dil into intact or resected liver was studied by fluorescence microscopy. The main structural changes in the kidneys after MSC injection into intact and partially resected liver manifested as granular dystrophy and necrobiotic/necrotic changes in single epithelial cells of the distal tubules and collecting ducts, thrombosis of some vessels, progression of an ascending urinary tract infection (detection of dust-like fluorescent objects), which can be due to the immunomodulating or even immunosuppressive influence of MSC and their detritus. MSC injected into the intact or resected liver, as well as the products of their degradation were not detected in the kidneys at all terms of observation. After injection of MSC into partially resected liver, manifestations of bacterial contamination of the renal medulla appeared later. The injection of MSC into the liver can be complicated by thrombosis of the renal vessels, which should be taken into account when using this administration route in the cell therapy.

Dissemination of Multipotent Stromal Cells in the Organism after Their Injection into Intact and Resected Liver in the Experiment.

The possibility of dissemination of bone marrow multipotent stromal cells stained with Vybrant CM-Dil after injection into an intact and resected liver was studied using luminescence microscopy. Labeled cells were found in the kidneys, spleen, lungs, axillary, mesenteric, and inguinal lymph nodes. We observed dissemination of multipotent stromal cells and their detritus throughout the body that occurred only after filtration in the lungs, where most cells underwent destruction. Perivascularly located macrophages in various organs can phagocytize multipotent stromal cells and their detritus from blood vessels. The content of objects labeled with Vybrant CM-Dil in distant organs was significantly lower after multipotent stromal cell injection into the resected liver, which was associated with the deposition of cells in the damaged area of the organ and their partial entry into the abdominal cavity.

[The cell technologies in modification of mesh materials used in urology].

Based on the literary analysis, the influence of cellular technologies on the results of implantation of mesh materials was studied. The scientific literature of recent years contains a large amount of data devoted to the study of mesh structures and the possibilities of their modification using multipotent stromal cells (MSC) for implantation into patients for correcting tissue defects and pelvic organ prolapse. However, the ideal implant has not yet been created. Additional studies with a longer follow-up period are needed to determine the most successful and safe methods and materials for the restoration of pathologically altered or lost tissues and the transition to clinical trials. It is also yet to come to an unambiguous understanding of the best sources of MSC, ways for stimulation of proliferation, preservation and delivery of these cells into the necessary tissues of the body, to thoroughly study the causes of inefficiency and the risks of developing various complications, especially in the long term. The progress of urological implantology in modern conditions, of course, will be associated with the introduction of modern materials and technologies, including the using MSC.

Preservation of Mesenchymal Stem Cell-Derived Extracellular Vesicles after Abdominal Delivery in the Experiment.

Light luminescent microscopy was used to study the distribution of extracellular microvesicles with PKH26-stained membranes secreted by placenta-derived mesenchymal stromal cells in the uterine tissues at different terms after injections to intact rats and after abdominal delivery (a model of cesarian section). Microvesicles migrated through the uterine tissues and were detected for at least 8 days after injection. In some cases, microvesicles were more numerous in the uterus after cesarian section modeling, which can be related to blockade of microcirculation and lymph flow due to inflammation accompanying surgical intervention. The content of microvesicles in the uterine tissues gradually declined due to macrophage phagocytosis and, probably, due to their migration into the vascular bed. Despite their size, properly stained extracellular microvesicles can be detected by light microscopy in tissues after injections.

[Liver regeneration after resection in clinical and experimental conditions]. / Nekotorye aspekty regeneratsii pecheni posle rezektsii v klinike i eksperimente.

The researches devoted to postoperative liver regeneration and influence in this process were analyzed. Liver injury is followed by hypertrophy of residual liver parenchyma. The use of various cytokines is perspective for activation, acceleration and inhibition of liver recovery. Cellular technologies in the treatment of liver diseases can affect its repair. Moreover, these methods could make unnecessary resection and transplantation of liver in certain cases. It is generally accepted that the main effect of multipotent stromal cells (MSC) in liver failure is associated with their differentiation to the cellular elements of this organ. At the same time, recent reports revealed that MSC injection to the liver is followed by their quick death, dissemination to other organs and tissues or even elimination from the organism. Regeneration of non-parenchymal structures (vascular network and bile ducts) should be considered in addition to functional recovery of liver parenchyma after resection. Clarification of indications and contraindications for MSC therapy, as well as prevention of possible complications associated with cellular technologies are required.

[Age-related changes of the female urethra.]

The minimum and maximum indicators of the length, width, area or volume of organs and structures in the lower urinary tract can normally vary up to 2-3 times. With age, at healthy women the absolute and relative length of the urethra, the urethrovesical angle, and the inclination of the urethra do not change. Both smooth and striated muscle tissues, which are part of various departments of the female urethra, undergo atrophy during the aging process. Smooth muscle tissue is less variable with age, but striated muscle symplasts are sometimes completely absent in urethral biopsies from elderly patients. With age, the vascularization and density of the innervation decrease in the urethral structures, but the content of connective tissue in the external urethral sphincter increases. Urinary tract mobility at young women is more pronounced than at older women. The apparent insufficiency of modern investigations about age-related changes in the lower urinary tract indicates the relevance and need to continue studying of the organ and tissue transformation in the urinary system during aging, this may be important for determining the etiology and pathogenesis of some pathological conditions, such as stress urinary incontinence, as well as for differential diagnosis between age norm and pathology.

[Current trends of the choice and processing of materials for dental implantation]. / Sovremennye tendentsii vybora i obrabotki materialov dlia dental'noi implantatsii.

For assessment of the modern situation about the choice of materials for manufacture of dental implants and the processing of their surface the scientific literature for the last 2 years was study. On the basis of a large number of contradictory results of the researches devoted to each of dental implantation problems it is possible to draw a conclusion that any of primal problems of implantology is finally not solved. There is no unique opinion at the choice of optimum material for manufacture of dental implants, at the way of processing and modification of their surface. The problem of improvement of quality of dental implantation and fight against complications of this procedure cannot be solved simple drawing other substances on the implanted material surface, this task more easily and more successfully is solved via changes of product structure and various modification of implant surface. Up to the present the researches of an opportunity to influence on characteristics of the implanted materials, changing their structure and character of a surface, continue. And the publications reporting about the considerable positive effect of artificially created roughnesses on product surfaces, and the articles claiming that there are no big differences between the rough and polished implants are confirmed by objective measurements with statistical processing of the obtained data. It should be noted that among articles there are very many works of the doubtful plan or with insufficiently valid conclusions. This review leads to the conclusion that further clinical and experimental studies and about the choice of materials for manufacture of implants and at the ways of processing of their surface are necessary.

Injection of Multipotent Mesenchymal Stromal Cells as a Cause of Hemorrhages in the Regional Lymph Nodes: Experimental Study.

Hemorrhagic changes after subcutaneous injection of autologous bone marrow multipotent mesenchymal cells with transfected GFP gene and additionally stained cell membranes to WAG rats in the projection of ligated femoral vein were studied by fluorescent microscopy. Hemorrhages in tissues with experimental acute local venous occlusion were caused by a combination of venous hypertension with inflammation around the foreign body - the ligature used for ligation of the vein. Fibrin found in tissues together with erythrocytes in the hemorrhages could stimulate the formation of granulations and new vessels instead of damaged or thrombosed ones. Multipotent mesenchymal stromal cells and their detritus getting into the regional lymph nodes initiated immune reactions morphologically confirmed by stubborn hypertrophy and hyperplasia of the lymphoid nodules, hemorrhages, and manifest diapedesis of erythrocytes to the organ parenchyma and sinus system.

Some Peculiarities of Local Distribution of Multipotent Mesenchymal Stromal Cells after Their Injection into Intact Muscle Tissue in Experiment.

Changes in the muscular tissue after subcutaneous injection of autologous bone marrow multipotent mesenchymal stromal cells transfected with GFP gene and additionally stained with cell membrane dye Vybrant CM-Dil in the projection of ligated femoral vein were studied by light microscopy with luminescence. Stromal cells injected through the skin can appear not only in the damaged tissue where acceleration of regeneration processes is required, but also in intact structures located in superficial or deeper layers. In intact muscular tissue, stromal cells spreading in the perivascular tissue initiate inflammation and migration of macrophages, activate and even trigger sclerotic processes due to differentiation into connective tissue cells (fibroblasts) and stimulation of proliferation and collagen synthesis by host fibroblasts. Injected multipotent mesenchymal stromal cells are gradually phagocytized by macrophages.

Hypolipidemic effect of mannans from C. albicans serotypes a and B in acute hyperlipidemia in mice.

Mannans, which are biological macromolecules of polysaccharide origin and function as immunomodulators, have been shown to stimulate macrophages in vivo by interaction with the mannose receptor. Thus, they can be used to stimulate macrophages in order to effectively remove circulating atherogenic lipoproteins. Our primary aim was to evaluate the hypolipidemic potential of mannans from C. albicans serotype A (mannan A) and serotype B (mannan B) in a murine model of hyperlipidemia. Mannan A and mannan B were shown to significantly (p<0.05) stimulate both the proliferation (p <0.05) and nitric oxide production of murine peritoneal macrophages in vitro. Pre-treatment of CBA/Lac mice with mannan A prior to induction of hyperlipidemia significantly (p<0.001) reduced serum atherogenic LDL-cholesterol, total cholesterol, and triglycerides. Mannan B exhibited a similar, but more potent, hypolipidemic effect. Electron microscopic analysis of liver revealed a significant (p<0.001) decrease in the volume of lipid droplets when hyperlipidemic mice were pretreated by both mannans. In conclusion, our findings would suggest that both polysaccharide-based biological macromolecules evaluated in the present study, specifically, the natural immunomodulators (mannans A and B), appeared to function as effective lipid-lowering macromolecules, which could potentially serve as adjunct therapy to more conventional hypolipidemic medications such as a statin drug.

Results of Experimental Ligation of the Main Vein with the Use of Cell Technologies.

Autologous multipotent mesenchymal stromal cells (MMSC) of bone marrow origin with transfected GFP gene and additionally stained cell membranes were injected to rats through the skin in the projection of ligated femoral vein. The results were evaluated by fluorescent microscopy. No signs of MMSC incorporation into the wall of ligated vessel or reorganized collaterals were detected. Angiogenesis processes involving MMSC were detected in experimental rats within just 4 days and progressed until week 2 postinjection, mainly in granulations at the site of surgical intervention and the cicatrix forming there. Injected MMSC completely formed all tunics of the new vessels and incorporated in the vessels forming from the recipient cells. MMSC and the objects created from them were gradually eliminated with participation of macrophages and replaced by structures formed from the recipient cells.

Correction to: Possibility of Aggravation of Tissue Sclerosis after Injection of Multipotent Mesenchymal Stromal Cells Near the Forming Cicatrix in the Experiment.

The author name G. A. Chastikina should read G. A. Chastikin.

Possibility of Aggravation of Tissue Sclerosis after Injection of Multipotent Mesenchymal Stromal Cells Near the Forming Cicatrix in the Experiment.

The peculiarities of tissue sclerosis after injection of autologous bone marrow multipotent mesenchymal stromal cells transfected with GFP gene and stained with Vybrant CM-Dil cell membrane dye were studied by light microscopy with luminescence. The surgical intervention consisting in ligation of the great vein was followed by tissue sclerotic transformation caused by direct damage and chronic inflammation caused by the presence of slowly resorbed ligature. Injection of stromal cells after this intervention led to formation of more extensive scar. This can attest to the possibility of stromal cells differentiation into connective tissue cells, fibroblasts, and stimulation of proliferation and collagen synthesis by host fibroblasts. A decrease in the volume of dense fibrous connective tissue due to scar reorganization at latter terms cannot not excluded.

Features of Blood Supply to Axillary Lymph Nodes in Breast Cancer Patients.

Numerical density of vessels in axillary lymph node structures in breast cancer patients was studied by light microscopy with antibodies to CD34. The correlation between the stage of breast cancer and vascularization of lymph nodes was analyzed. The development of breast cancer was followed by rapid and significant increase in the number of blood vessel in axillary lymph nodes in general and in all their specific areas, especially in the paracortex and medullary substance. Strong or very strong positive correlation between the stage of tumor process and parameters of vascularization was observed in various areas of the axillary lymph nodes. No significant differences in node vascularization were observed between women without breast cancer and women with stage I breast cancer; the differences were found only starting from stage II. Considerable vascularization of lymph nodes can be a symptom of malignant tumor growth in the area of lymph collection. The duration of the tumor process and the size of the tumor can be indirectly evaluated by the increase in vessel number in lymph nodes.

Initial Stages of Angiogenesis after Acute Experimental Local Venous Outflow Disturbances and Application of Cell Technologies.

The initial stages of angiogenesis in rats after transcutaneous injection of autologous bone marrow multipotent mesenchymal stromal cells transfected with GFP gene and stained cell membranes in the projection of ligated femoral vein were studied by fluorescent light and confocal laser microscopy. Large clusters of brightly fluorescing elongated fibroblast-like cells were seen in the paravasal tissue and in the postoperative scar and signs of angiogenesis were noted as soon as in 4 days. The injected cells not only formed new vessels, but also integrated into vessels formed by host cells. Some injected cells were phagocytizied by macrophages and the latter started to fluoresce due to the presence of the membrane dye. These macrophages within the specified period appeared in the regional inguinal lymph nodes where they formed clusters in the lymphoid parenchyma of the cortical substance.

Possibility of Using Mesenchymal Stromal Cells to Restore Lymph Flow in Experimental Phlebothrombosis.

The possibility of formation of lymphatic vessels after introduction of autologous bone marrow-derived multipotent mesenchymal stromal cells transfected with GFP gene into thrombosed femoral vein was studied by fluorescent microscopy. Vascular thrombosis caused by ligation of the great vein with subsequent injection of thrombin solution was accompanied by blockade of regional lymph flow. The cells injected into thrombosed vein directly participate in the formation of new lymphatic vessels in the paravasal tissue surrounding the vein, its tissue region, and around regional lymph nodes. This is seen from bright specific fluorescence of individual cells in the walls of lymphatic vessels and all vascular layers and valves in UV light.

[SOME REACTIONS OF THE REGIONAL LYMPH NODES OF RATS AFTER IMPLANTATION OF MULTIPOTENT STROMAL CELLS ADSORBED ON POLYHYDROXYALKANOATE INTO A BONE TISSUE DEFECT].

The reactions of the regional lymph nodes, caused by implantation of the autologous multipotent stromal cells of bone marrow origin (AMSCBMO) to accelerate the healing of mandibular bone defect were studied by fluorescent microscopy in inbred male Wag rats aged 6 months (n=62). After the introduction of polyhydroxyalkanoate transplant containing adsorbed AMSCBMO with a transfected Green Fluorescent Protein (GFP) gene into a damaged bone area, the lymphoid nodules in submandibular lymph nodes demonstrated the appearance of numerous large macrophages containing multiple oval fluorescent inclusions in the cytoplasm. The number of these macrophages increased within 2 weeks after surgery and then began to decline. Apparently, AMSCBMO introduced in this way, were partially absorbed by macrophages. After destruction of the structures formed from AMSCBMO, the debris was also phagocytized by macrophages. In either case, these macrophages appeared in the germinal centers of lymphoid nodules in lymph nodes, where the induction of immune responses against DNA and GFP protein was probable.

[The age changes of lymphoid organs structure: review of literature].

Numerous and often contradictory research results specify that the mechanism of increased susceptibility to diseases in old age has no satisfactory explanation so far. Together with it in literature almost completely there are no comparative data on features of surgery and injury influence on a structure and functions of lymphoid organs at patients of different age groups. However, lymph nodes are a marker of activity of inflammatory process in the region, on their changes it is possible to estimate precisely productivity of different medical actions, to predict development of many complications, and, thus, successfully to take actions for their prophylaxis. All this is especially important and actual because of constant increase in average age, in life expectancy of human population and strengthening of surgical activity at patients of advanced and old age.

The features of lymph node reactions to tissue damage of lymph collecting region at old rats.

The features of submandibular lymph node (LN) reactions after lower jaw tissue damage at old rats were studied by method of light microscopy. In LN of old rats for all terms of observation, including intact control, the volume of connective tissue is considerable increased. Besides, old animals in LN have of more change duration of a cortical plateau volume, less - of paracortex and lymphoid nodules without light centers. Features of cellular element number consisted in considerable decrease in quantity of lymphocytes, mitotic figures, immunoblasts and plasmablasts against increase in keeping of macrophages, cells with destruction, stromal elements and erythrocytes.

[ANGIOGENESIS IN THE TISSUES AFTER THE INJECTION OF STROMAL STEM CELLS OF BONE MARROW ORIGIN CLOSE TO THROMBOSED VEIN IN AN EXPERIMENT].

The effects of the injection of autologous multipotent stromal stem cells of bone marrow origin (MSSCBM) (mesenchymal stem cells) with green fluorescent protein gene, additionally marked with DAPI nuclear stain, close to a thrombosed hindlimb vein, were studied by fluorescent microscopy in adult male Wag rats (n = 214). The control groups consisted of intact rats (n = 12), animals with venous thrombosis without the injection of MSSCBM (n = 71) and rats that received paravasal injection of MSSCBM, but without preliminary modeling of venous thrombosis (n = 72). It was found that MSSCBM participated in the development of granulation tissue at the site of surgical intervention performed during the modeling of thrombosis. The rapid development of granulation tissue at the site of surgical trauma may contribute to faster wound clearance from detritus, nonviable tissue and antigenic substances, early onset of tissue repair processes and rapid healing. Restoration of blood flow in the tissue region of a thrombosed vein began later than after the intravenous injection of MSSCBM.