[Wounds treatment by the transplantation of the fibroblasts and stem cells on collagenic matrix].

During many decades the development of effective methods of wounds treatment has been and still remains to be one of the most actual problems of surgery. Interest and constant attention to wounds speaks first of all that simultaneously with development of medicine, cellular and molecular biology, molecular genetics and immunology consideration of wounds processing is constantly vary. New prospects and opportunities of effective treatment of wounds are opening. We develop a new original method of treatment full-thickness wounds by a method of transplantation fibroblasts and stem cells on the basis of collagenic matrix. The presented method of treatment of full-thickness wounds provides the most perfect realization of stimulating action of cultures of cells on the processes of regeneration of tissue. Application of fibroblasts and stem cells on the basis of collagenic matrix promotes restoration of normal structure of a skin and allows reducing terms of healing considerably. Stem cells are integrated into three-dimensional tissue structures of an organism and promote histotopically to restoration of defects of a skin covering. Here it is necessary to note, that transplanted skin fibroblasts synthesize and allocate in an environment a plenty of biologically active substances among which distinguish epidermal growth factor (EGF), fibroblast growth factor (FGF), transforming growth factor (TGF), keratinocyte growth factor (KGF), and also various components of extracellular matrix.

[Treatment of obstruction of extrahepatic biliary pathways by antireflux, bilio-cholecystic (cystico) enteroanastomosis methods].

The main target of our investigations was the development of antireflux, bilio-cholecysto (cystico) enteral anastomosis method for the surgical treatment of extrahepatic biliary duct obstruction. We were using the cholecystic duct spiral valve (Lutkens valve, Haister valve) as the antireflux structure. In this case the bile drains into the biliary cyst not through the cholecystic duct but through the extrahepatic biliary duct - the bilio-cholecystic anastomosis. Such an interposition of cholecystic and spiral valves promotes inhibition of chemical reflux between extrahepatic ducts and gastrointestinal tract, from digestive tract into the cholecystic and intrahepatic biliary tracts. Method developed and tested by us involves creation of blio-cholecystic (cystico) entero anastomosis, namely creation of biliary duct-cholecystic (cystico)-duodenoanastomosis. 3 successful experimental operations have been performed using the above presented approach. Sutures were taken of on the seventh day, animals were housed under investigations for 10-14 days, faecal masses had normal appearance.

[Treatment of thrombophlebitis by the method of the transplantation of the endothelial cells].

The end XX of century was marked by introduction into the clinical practice of the methods of cellular therapy. Intensive studies on its application for treating the whole series of diseases are conducted. Thrombophlebitis is not an exception in spite of the already existing effective surgical and conservative methods of its treatment. On the rabbits we produced experiments on the treatment of thrombophlebitis of "aseptic" and infectious etiology. The results of complex treatment with the use of autologous vascular endothelial cells were studied. The damaged wall of vein under the conditions of the "aseptic" model of thrombophlebitis, with conducting active complex therapy with the start of antibiotics and trombolitic means is completely covered with endothelium, and only to the place where the dust devils were attached to the walls of vein, defects remain. In these places the wall of vein gradually is extended and undergoes scarring, and the intact sections of vein around the defect are hypertrophied and then they undergo dilatation, being adapted thus to the new conditions of functioning. The transplanted endothelial vascular cells not only modify the process of remodulation of the damaged vein and thus they contribute to averting the dilation of its wall and expansion of scar, but also contribute to the retention of the thickness of the wall of vein. Calculations showed that in the case of "aseptic" etiology the processes of thrombogenesis and treating thrombophlebitis continue more rapidly than in the case of the infectious etiology. The comparison of traditional method of the treatment of thrombophlebitis with the complex (with the application of auto of endothelial cells) showed that the latter is more effective.

[New solution for cells and organs tissue conservation].

One of the major problems in organ and tissue transplantation is the conservation of donor's material from the moment of its impressments till its grafting in the recipient's organism. The new direction in transplantation is a method of liver tissue micro-fragments transplantation in bio-container Kakabadze, which was elaborated at the Department of Clinical Anatomy of Georgian State Medical Academy. In this work the results of investigation of hepatocytes' viability in micro-fragments were studied. The micro-fragments were placed in different medias, including the one elaborated by us on the bases of natural sugar-containing preservative. The decrease of percentage of viable hepatocytes in dependence of preservation time in different media was investigated. In all cases the dependence can be approximated by exponential function. On the bases of experimental data semi-empirical mathematical model of processes taking place in liver tissue micro-fragments during their preservation was created. Two parameters are introduced: one of them A depends on the method of micro-dissection, and the other T (1/2) is characteristic of the media and represents the time, in which the percentage of viable hepatocytes halves. The percentage of nonviable hepatocytes and also the percentage hepatocytes, which were in condition of apoptosis, were investigated. The investigations had shown that in all cases 50% barrier did not exceed 17 hours. This barrier in solution that is elaborated by us reached 46, 4 hours. Besides, if we assume, that the half of hepatocytes, which were in apoptosis condition, will become viable after transplantation, then the barrier will run up to 125 hours. The investigations of intra-cell glycogen showed that in all medias we had used, resources of glycogen were exhausted during one hour. In the case of solution elaborated by us glycogen was maintained in cells during 7-8 days.