IMMUNOMODULATORY EFFECT OF ALLOGENEIC MESENCHYMAL STEM CELLS OF RATS.

INTRODUCTION: Recently, more and more attracted the attention of cell therapy, which requires a study of the efficacy and safety of allogeneic MSCs transplantation in acute and chronic inflammatory reactions. The aim of our study was to examine the effectiveness of transplantation of allogeneic mesenchymal bone marrow stromal cells for the healing of surgical wounds the glandular stomach in rats. MATERIAL AND METHODS: Using white Wistar rats. Producing cell transplantation mononuclear fraction derived from rat bone marrow aspirate. Injected cells 8 and 9 th passage. The dose of cells administered to 3-th and / or 7-th days 3,5h106 cells / ml twice or 5,0h106 cells / ml dose. Autopsy on day 10-th and 17-th. The serum ELISA determined the content of the pro- and anti-inflammatory cytokines IL1ß, TNFα, IFNy, IL-4. RESULTS AND DISCUSSION: Introduction MSCs contributed to the decline of the immune mediators of inflammation IL1P, TNFa, IFNy, increase anti-inflammatory IL4. Quality improved healing. CONCLUSION: Rapid curative effect of stem cells may be associated with the formation of blood immune cells (macrophages) that produce substances that restore damaged tissue. They restore the balance between Th1 and Th2.

Cytogenetic recapitulation, induced by medical preparations, as the universal stage of formation of urgent protection against damage at organ transplantation.

In this article modern representations about cellular mechanisms of formation by pharmacological preparations of urgent protection of organs against damage are given at transplantation. On an example of ischemic damage of kidneys it is shown, that at use of preparations of different pharmacological groups by the most expressed protective effect those from them which operating within the limits of a nonspecific adaptive syndrome of cellular systems, clearly induce in organs the evolutional developed signs of cytogenetic recapitulation possess: support a cellular homeostasis at the lowered level at the expense of activation of a glycolysis and conformational reorganizations of macromolecules, and also change in cells of water contain- decrease of free and increase of bound.

[Correction of cronic liver failure by transplantation of liver cells suspension and cell-engineering designs (experimental investigation)].

On an experimental model of chronic fibrotic liver damage (male rats Wistar (n-60), damage of CCl4, the duration of the experiment 90 days) it was studied the effectiveness of cell therapy for the correction of chronic liver failure. These rats were divided into 3 experimental groups: in the Ist-group (control, n=10) isotonic saline (650 mkl.) was injected; in the IInd-group (n=20) suspension of liver cells was applicated in a dose 8 - l0 x 10(6) cells; in the IIIrd-group (n=30) suspension of liver cells and bone marrow cells (mesenchymal stromal cells) in ratio 5:1 were used as cell associates on microparticles intjectable heterogeneous biopolymer hydrogel "SpheroGEL" (cell-engineering design) in common dose 8 - l0 x 10(6) It was ascertained that in the 2nd and in the 3rd groups the accelerated normalization of disturbed liver functional indices (ALT, AST, ALP) took place - to 30 days, but in the control group only to 90 days. The reliable differences in rats ofnormalization offunctional indices were absent between the IInd and the IIIrd groups. But in 90 days by using special histological dyeing it was found out that defibrotic processes in liver tissue were more expressed in the IIIrd group in comparison with the IIIrd group. Received results were consequence of prolonged vital activity of cells (liver cells and mesenchymal stromal bone marrow cells) into cell-engineering designs, which were transplanted in the IIIrd group. The obtained effect can be explained by that the developed cell-engineering designs provide adequate conditions for prolonged vital activity of the transplanted cells.

[Mesenchymal stem cells transplantation in the treatment of radiation skin lesions].

Transplantation of mesenchymal stem cells, both at early and later stage after local exposure of rats source beta radiation dose, 90Sr/90Y 140 GR, stimulates recovery of damaged skin. Diminution area local radiation injuries and accelerate healing radiation ulcers. Clinically shows the high efficiency of the transplantations autologous mesenchymal stem cells in treatment of deep beam ulcers, intractable standard conservative treatment. Found promising application of mesenchymal stem cells for treatment of severe local radiation injuries and the need to develop the best possible conditions for their use.

[The cultivation of bone marrow cells and cell lines on polymeric films].

The cultivation of multipotent mesenchymal stromal bone marrow cells and cells of A-431, MDCK, Vero, 3T3 and Hep-G2 was performed on polymeric films (PVA) with different hydrophobic fatty acid residues. The cells of different types grew on these films with different intensity, but in the most cases comparable with the cultivation control on usual plastic. The examined films were nontoxic to cells and sufficiently adhesive. They did not changed pH of cultural media, were optically transparent under microscope and comfortable in the experimental work. These films can be used as a model for the artificial organ construction. The covalent binding of different fatty acids to PVA shows possibility of the adaptable changes of films properties (hydrophobity and adhesiveness), and therefore possibility of the creation of optimal conditions for different cell types attachement and growth.

[Cytokine imbalance correction and regeneration stimulation of persistent autoimmune stomach ulcers with help of periulcer transplantation of mononuclear cells of autological bone marrow].

The efficiency of correction of cytokine disbalance and peptic ulcer regeneration has been studied using the model of chronic peptic ulcer at use based on mononuclear fraction cells (MFC) of autologous bone marrow (BM). MFCs are shown to remove cytokine disbalance, reduce systemic inflammation reaction and accelerate peptic ulcer regeneration.

[Multipotent mesenchymal stromal cells of the autologous bone marrow accelerate healing indolent gastric ulcers].

The influence of cultivated multipotent mesenchymal stromal cells (MMSC) of the autologous bone marrow on activity of wound healing processes was studied using the model of long-indolent autoimmune gastric ulcer in rats. MMSC of the bone marrow when under the state of stress inhibition do not possess optimal and regulatory activity. Precultivated MMSC of the bone marrow accelerate the processes of regeneration of the gastric ulcer. MMSC of the bone marrow realize their bioregulatory

[Use of precultivative multipotent mesenchymal stromal cells isolated from autologic bone marrow in treatment of non-healing autoimmune gastric ulcers].

It is shown that multipotent mesenchymal stromal cells (MMSC) grafted to the zone of ulcer defect contribute to better wound repair in case of chronic non-healing gastric ulcers. It is noted that precultivative MMSC isolated from bone marrow achieve their bioregulatory activity in patient's organism not at once but due to prolonged functioning in the grafting zone (not less than 30 days).

[Regenerative cell therapy of the type I diabetes mellitus and its complications].

This review describes up-to-date concepts of the type I diabetes mellitus pathogenesis and its complications. The leading role of the immune system malfunction in pancreatic islet of Langerhans and in vessel wall regeneration impairment is emphasized. It is suggested, that cell therapy of the type I diabetes mellitus with pancreatic islet and bone marrow cell transplantation promote beta-cell regeneration due to normalization of intracellular interaction in these tissues and immune homeostasis in whole organism.

[Improvement of myocardial remodelling and functioning in patients undergoing surgical treatment of chronic cardiac insufficiency by two-staged elevation of the immunoregulatory reserve of autological bone marrow cells and their intramyocardial application].

The aim of the investigation was to study a possibility to improve left ventricular (LV) volume and function remodelling in patients with chronic cardiac insufficiency (CCI) by means oftwo-stage bone marrow cell (BMC) activation: first in vivo and then ex vivo within the process of their cultivation. Two groups of CCI patients were recruited. Group 1 (controls) consisted of 50 patients undergoing conventional aorto-coronary bypass surgery (ACBS). Group 2 consisted of 57 patients injected with 200 million autological mononuclear BMC intramyocardially during ACBS. In Group 2 patients, the severity of immune dysregulation was assessed initially using blood leukocyte stimulation index (SI) values. Sixteen patients with SI >1 and moderate immunographic alterations were considered to have a favorable prognosis for BMC treatment; 41 subjects with SI <1 and pronounced immunographic abnormalities were regarded as having an unfavorable prognosis for BMC application. Twenty-two patients with SI <1 were administered a preliminary immunocorrective course (in vivo BMC activation). Mononuclear BMC obtained from 38 patients with SI >1 and 19 patients with SI <1 were cultured for 5 to 7 days (ex vivo BMC activation). Positive changes in BMC phenotypical pattern were observed only in patients with SI >1: CD3, CD4, CD8, CD25, and some other measurements increased. Significant positive effects on LV function parameters and Duke Activity Status Index (DASI) values were revealed in patients with SI >1 six months after BMC administration. In vivo immunocorrection in combination with subsequent ex vivo BMC activation (n=38) promoted significant improvements in LV volume characteristics 6 months after ACBS vs. the controls (ACBS without BMC, n=50). In conclusion, to make the administration of autological BMC in CCI patients effective, two-staged BMC activation should be performed: in vivo activation with immunocorrectors should be followed by ex vivo activation of cultured cells.

[Diabetic complications in rats in long-term modeling of type I diabetes mellitus].

Diabetes mellitus type 1 (DM-1) was modeled by a single intraperitoneal injection of streptozotocine (STZ) in a dose 65 mg/kg in two groups of Wistar rats: control (n=6) and test (n=14). Clinical and pathomorphological examinations of different organs were conducted for 4.5 months. It was found that the above model represents clinical picture and vascular diabetic complications (microangiopathy with parenchymal lesions) accompanied with destruction and depression of spleen activity. It is shown that rats, more resistant to the toxic action of STZ restore the ability for regeneration of the pancreatic islets and regress of glycemia and angiopathy, for positive changes in splenic structure and function.

[Controlled production of active oxygen forms by leukocytes allows prognosis of the body resistance to stress damage].

The authors suggest using chemiluminescent reaction of the patients' blood leukocytes including the stimulation index (SI) for prediction of the body resistance to stress damage (operative stress). It is shown that in chronic cardiac patients (n = 259) SI is initially subnormal. Some of the above patients had critically low SI ( > 0.6). Such patients developed severe immunodependent postoperative complications. To increase the patients' resistance to operative stress and to avoid postoperative complications it is recommended to conduct prophylactic immunocorrecting therapy under SI control.

[Transposition of autologous cultivated bone marrow cells promotes prevention and treatment of immunodependent complications in cardiosurgical patients].

Coronary artery bypass grafting (CABG) has been made in two groups of patients with chronic heart failure (CHF) with further estimation of the rate of postoperative organic dysfunctions and pyoseptic complications. In group 1 (n = 50) CABG was combined with intracoronary or intramyocardial injection of autologous precultivated for 7-8 days mononuclear cells of the bone marrow (1 x 10(9) cells). In group 2 (n = 479) the intraoperative injection of the above cells was not made. It was found that autologous cultivated mononuclear bone marrow cells prevent organic dysfunction and reduce frequency of infectious-septic complications especially in patients with preoperative focuses of chronic infections.

[Inflammation and apoptosis in relation with the effectiveness of bone marrow cell transplantation to patients with chronic heart failure].

The authors analyze the pathogenetic significance of inflammation, immune activation, and apoptosis in the development of heart failure and the functional regeneration of left ventricular myocardium after treatment including autologic bone marrow cell transplantation. The paper shows that autologic bone marrow cell transplantation influences the same mechanisms that cause heart failure, namely inflammation, apoptosis, and neurohumoral mechanisms. Investigations have made it possible to formulate guidelines for the prediction and monitoring of the effects of treatment including autologic bone marrow cell transplantation in patients with heart failure.

[Cell therapy as a method to correct pathogenetic disturbances in dyslipidemia and early atherosclerosis].

The article is dedicated to the pathogenetic mechanisms of the development of immune inflammation in the vascular wall, as well as hepatic functional disregulation in dislipidemia and atherosclerosis. The authors emphasize that the decompensation of the process of cell regeneration in actively functioning organs (vascular endothelium, hepatocytes) and a concomitant immune disregulation become the main factors of the progression of dislipidemia and atherosclerosis. The authors consider the conditions for and factors of cell therapy, based on the use of allogenic fetal hepatic cells and autological bone marrow cells.

Cell transplantation inhibits inflammatory reaction and stimulates repair processes in burn wound.

We compared the effects of transplantation of fetal fibroblasts and fibroblast-like mesenchymal stem cells of the bone marrow on healing of deep burn wound in rats. It was found that transplantation of fetal fibroblasts and fibroblast-like mesenchymal stem cells on the burn surface reduces cell infiltration, promotes the formation of vessels and granulation tissue, which creates conditions for more rapid healing of the burn wounds.

Use of autologous bone marrow mesenchymal stem cells for healing of free full-thickness skin graft in a zone with pronounced hypoperfusion of soft tissues caused by arteriovenous shunting.

A combined graft consisting of a free full-thickness skin flap and cultured autologous fibroblast-like bone marrow mesenchymal stem cells was effectively implanted and healed on the facial soft tissue defect after removal of a pathological vascular conglomeration in a female patient with congenital arteriovenous macrofistulous dysplasia. In order to reduce bloodflow intensity and arteriovenous shunting, repeated endovascular occlusion and transcutaneous ligature of regional vessels from the carotid artery basin feeding the pathological zone was carried out followed by resection of this tumor-like vascular formation.

Use of recombinant AdSV40-BetaGal adenoviral construction for monitoring of transplanted cells.

Original recombinant adenoviral construction carrying E. coli beta-galactosidase LacZ gene designed by the authors is convenient for labeling and monitoring of bone marrow mesenchymal (stromal) progenitor cells and myocardial and skin fetal cells transplanted in damaged rat tissues (in the perinecrotic zone of the myocardium and onto burnt skin surface) for their reparation. This genetic construction after pre-inactivation of endogenous beta-galactosidase allows to detect transplanted cells in the foci of injury; positive effects of transplantation on tissue reparation processes can be attributed to the presence of transplanted cells.

ElastoPHB membrane systems with immobilized bone marrow stromal cells optimize conditions for regeneration of damaged tissue.

The effects of autologous bone marrow stromal cells immobilized on ElastoPHB membranes on reparative processes were studied on a model of rat skeletal muscle injury. Bone marrow stromal cells inhibited substitute (sclerosing) regeneration and activated reparative (reconstructive) regeneration of tissues.