Морфологічне дослідження ефективності застосування клітинних технологій у корекції експериментальної виразки шлунку

Introduction. Modern methods of treatment of gastric ulcers, both conservative and surgical, aimed at creating an optimal balance between the factors of "defense" and "aggression" mucosa, leading to the healing of the ulcer. However, these approaches are not in any way affect the actual on reparative processes in the wall of the stomach, which adversely affect the course of disease in general. Solving this problem requires going beyond traditional methods of treatment. The most promising areas is the use of plasma-rich platelets as a source of growth factors or autologous purified lipoaspiratu as donor mesenchymal stromal stem cells polipotentnyh. Objective: to evaluate the morphological features of healing of experimental gastric ulcer lesions in terms of the use of autologous purified lipoaspiratu and plasma enriched with platelets. Materials and methods. Experimental research conducted at the vivarium Vinnitsa National Pirogov Medical University. All experiments were performed according to the "Regulations on the use of animals in biomedical experiments" with the permission of the Bioethics Committee. To study included 48 white laboratory rats of both sexes under the age of 1 year and weighing 120 to 220 g (186 ± 17 g). The control group consisted of 12 rats, which were modeled on symmetric ulcerative lesions of the stomach. The other two groups included 18 rats with simulated gastric and symmetrical subsequent stimulation of reparative processes in the gastric anterior wall of the stomach. 1 experimental group consisted of rats WHO to accelerate the repair of locally injected autologous purified lipoaspirat 2 research group - rats, for whom locally injected plasma enriched with platelets. Modeling symmetrical ulcerative lesions of the stomach by a modified method Susumu Okabe using the author's device was made all 48 rats. Assess the state of existing ulcers was performed 3 days after modeling when (according to published data) completely formed ulcerative defects. To do this selectively picked 4 rats from each of the 3 groups. Macroscopically assessed the state of the walls of the stomach and abdomen. Then vivo measured area formed ulcers front and rear walls of the stomach on the author's technique. After fixing performance areas ulcers animals taken out of the experiment by dislocation of the cervical vertebrae. Removed the stomach, gastrotomy performed and evaluated macroscopically condition mucosa. Material taken for further morphological studies. The experimental results evaluated at 7 and 14 days from the date of repair stimulation - measured performance areas ulcers, and took away material for morphological studies. To evaluate the pathological changes of gastric mucosa experimental fixed in 10% formalin solution neutral. Preparations were prepared by the standard method, histological sections 5-7 microns thick were stained with hematoxylin, eosin, Sudan III, pikrofuksynom by Van ghisoni main brown for Shubichem, a combination of brown and strong core of green dye with PAS reaction altsianovym blue. Microscopy and histological preparations photographing performed using a light microscope Olympus BX 41 at magnifications of 40, 100, 200 and 400 times. Results. Histological analysis of acetic ulcer after 7 days of the experiment revealed preference destructive inflammatory changes in the gastric mucosa. Thus, the surface epithelium of the mucous membrane in the area of the ulcer was necrotic. Microscopic analysis reveals the shallowness characteristic at a time of observation: the outer layer at the bottom and edges of the ulcers presented fibrinous-purulent or purulent exudate. Followed by fibrinoid necrosis zone, which is located deeper layer of granulation tissue, and even deeper - fibroblasts. At the bottom of the ulcer manifested structureless mass of desquamated epithelial cells limfotsytodopodibnyh impurities. When necrotic areas were observed among granulation tissue, small intermittent zones of fibrinoid necrosis. In experimental groups, where stimulation of reparative processes performed using lipoaspirat and plasma enriched with platelets, ulcers on their morphological structure on the seventh day were similar to the acetate ulcer: ulcers proximal end hung above its crater, distal end smoothly passed to the mucosa. Layers of purulent exudate and fibrinoid necrosis turned into the lumen of the stomach, beneath formed granulation tissue. In rats injected plasma enriched with platelets, the depth and degree of inflammatory infiltration compared with the acetate ulcer without correction was significantly lower (0,189±0,010±11,45 and 407 respectively, p<0,001). 14 day experiment in rats with acetic ulcer without correction biological stimulants progressed and ulcerative defect healing was observed in any animal. Pathomorphological changes acetate stomach ulcers not significantly different from the 7 days and were characterized by the persistence of floors According to the morphometric study average depth of ulcers in the group lipoaspiratu use in this study period was 0,155±0,007 mm. In the group of rats injected plasma, platelet-rich epithelium holes 14 day experiment, by contrast, was generally healthy appearance. The cytoplasm of its cells filled with moderately secret basophilic, indicating that a large number rybonukleyidiv. There were figures mitotic division. Proliferation of cells in the deeper parts of the pits led to their extension and acquisition of tortuous or shtoporopodibnoho look. In some animals the use of plasma in the area of epithelial defect observed focal intestinal metaplasia. In the group of experimental animals injected lipoaspirat complete epithelization ulcer was observed in 2 (28.6%) cases, platelet-rich plasma - 5 (71.4%). In the group of acetate ulcers not healed ulcer none during the experiment. Morphometric study found that 14 days of the experiment the number of macrophages virtually unchanged in all study groups, but the number of fibroblasts in the case of plasma (209 ± 18,10) numerically higher than obtained in the group of acetate ulcers without correction. The size of the plasma cells and lymphocytes had the opposite trend and characterized their advantage in the group of acetate ulcers without correction, which can prolong suggests chronic inflammation in these rats. The results of morphological studies of acetic acid gastric ulcers healing in rats with correction of the reparation process with local use of autologous purified lipoaspirate and platelet rich plasma are shown in this article. Platelet rich plasma significantly better than lipoaspirate reduces inflammatory response and stimulates proliferation of gastric epithelial cells on 7th day with the restoration of secretory activity and epithelialization of ulcers in 71.4% of experimental animals on 14th day, and the activation of the fibroblastic reaction during the all experiment. Conclusions. For the purposes lipoaspiratu morphological changes after 7 and 14 days of the experiment did not provide decongestants and anti-inflammatory effects, but the depth of the ulcer and the degree of inflammatory changes was significantly lower compared with the acetate ulcer without correction (p<0,001), and epithelization 14 day experiment 28.6% was determined in experimental animals. The plasma enriched with platelets, reduces the inflammatory response and gastric epithelial cells stimulates proliferation on day 7 of the restoration of secretory activity and epithelialization of ulcers in 71.4% of experimental animals at 14 days, and the activation of the fibroblastic reaction during the entire duration of the experiment.