Morphological and morphometric changes on the background of cell cardiomyoplasty in experimental myocardial infarction

S. Estrin, T. Kravchenko, A. Pechenenko
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Abstract

The aim: to study the morphological and morphometric changes in the myocardium against the background of cellular cardiomyoplasty in experimental myocardial infarction. Materials and methods: the experiment was carried out on 142 Wistar-Kyoto rats weighing 200–220 g, which were kept in the vivarium of the Department of Experimental Surgery of the State Institution “Inst itute of Emergency and Reconstructive Surgery named after V. K. Gusak of the National Academy of Medical Sciences of Ukraine” in the period from 2012 to 2013. The Wistar-Kyoto breed was used because it is inbred, which minimizes the rejection reaction, due to its genetic homogeneity. The animals were kept in a vivarium under conditions of 12-hour daylight hours, room temperature and access to water and food at libitum at an air temperature of +20 – + 22 °C, humidity no more than 50 %, in a light mode day-night. The use of animals in the experiment was carried out in accordance with the rules regulated by the “European Convention for the Supervision and Protection of Vertebrate Animals used for Experimental and Ot her Scientific Purposes” (Strasbourg, 1986), Directives of the Council of the European Union of November 24, 1986 and the order of the Ministry of Health of Ukraine No. 32 dated 02.22.88. The induction of my ocardial infarction (MI) was carried out according to the technique developed by us under general anesth esia. A separate group consisted of 20 males, whom we used as donors of mesenchymal stem cells (MSC) for further research on the Y chromosome of cell homing in the body. Cultivation of MSCs was carried out in a mixture of nutrient media DMEM / F12, 1:1, (Sigma, USA). The material for morphological studies was the sections of the myocardium of laboratory animals. To assess the morphometric parameters, histochemical methods were performed according to the recipes, which are given in the instructions for histochemistry. Immunohistochemical study was performed on paraffin sections with a thickness of 5-6 μm by the indirect Koons method according to the Brosman method (1979). Results: it was found that cellular cardiomyoplasty significantly improves the structure of the postinfarction heart, manifests itself in a decrease in the scar area and connective tissue, respectively, in an increase in the number of vessels and the percentage of preserved muscle fibers. The best results were achieved with intramyocardial injection, which requires confirmation of this fact in a clinical study. Conclusions: cellular cardiomyoplasty with any method of introducing a cell graft has a positive effect both on the morphological substrate of the heart in the form of a decrease in the size of the scar during postinfarction remodeling, an increase in the number of newly formed vessels and an increase in the percentage of preserved cardiomyocytes. This occurs due to the homing of MSCs into the ischemic zone and the commonality of two mechanisms – direct differentiation into endothelial cells of the heart vessels, as well as due to the paracrine effect
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实验性心肌梗死细胞心肌成形术背景下的形态学和形态计量学变化
目的:研究细胞心肌成形术对实验性心肌梗死心肌形态学和形态学的影响。材料与方法:实验选用Wistar-Kyoto大鼠142只,体重200-220 g,于2012 - 2013年饲养于“乌克兰国家医学科学院急诊与重建外科研究所”实验外科实验室。使用Wistar-Kyoto品种是因为它是近亲繁殖的,由于其遗传同质性,这可以最大限度地减少排斥反应。饲养条件:白昼12小时,室温,自由取水和食物,温度+20 ~ + 22℃,湿度不超过50%,昼夜光照模式。在实验中使用动物是按照《欧洲监督和保护用于实验和科学目的的脊椎动物公约》(1986年,斯特拉斯堡)、1986年11月24日欧洲联盟理事会指令和1988年2月22日乌克兰卫生部第32号命令所规定的规则进行的。我的心梗(MI)的诱导是根据我们开发的技术在全身麻醉下进行的。另一组由20名男性组成,我们将其作为间充质干细胞(MSC)的供体,用于进一步研究细胞在体内归巢的Y染色体。在营养培养基DMEM / F12 1:1 (Sigma, USA)的混合物中进行MSCs的培养。形态学研究材料为实验动物心肌切片。为了评估形态计量学参数,根据组织化学说明书中给出的配方进行组织化学方法。参照Brosman法(1979),采用间接Koons法对5-6 μm厚度的石蜡切片进行免疫组化研究。结果:细胞心肌成形术显著改善梗死后心脏结构,瘢痕面积和结缔组织分别减少,血管数量增加,肌纤维保存比例增加。心内注射取得了最好的效果,这需要在临床研究中证实。结论:任何引入细胞移植物的方法的细胞心肌成形术对心脏的形态学基质都有积极的影响,其形式是在梗死后重塑期间疤痕大小的减少,新形成的血管数量的增加和保存的心肌细胞百分比的增加。这是由于间充质干细胞归巢到缺血区以及两种机制的共性-直接分化为心脏血管内皮细胞,以及由于旁分泌效应
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